MedWorm.com provides a medical RSS filtering service. Over 6000 RSS medical sources are combined and output via different filters. This feed contains the latest items from the 'Current Protocols in Neuroscience' source. Tue, 29 Nov 2011 20:04:01 +0100 http://www.medworm.com/index.php?rid=5297413&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D21971846%26dopt%3DAbstract Authors: Potter H, Heller R Abstract Electroporation-the use of high-voltage electric shocks to introduce DNA into cells-can be used with most cell types, yields a high frequency of both stable transformation and transient gene expression, and, because it requires fewer steps, can be easier than alternate techniques. This unit describes electroporation of mammalian cells, including ES cells for the preparation of knock-out, knock-in, and transgenic mice. Protocols are described for the use of electroporation in vivo to perform gene therapy for cancer therapy and DNA vaccination. Also described are modifications for preparation and transfection of plant protoplasts. Curr. Protoc. Neurosci. 57:A.1E.1-A.1E.11. © 2011 by John Wiley & Sons, Inc. PMID: 21971846 [PubMed - in proc... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=5297413 Sat, 01 Oct 2011 04:00:00 +0100 5297413 http://www.medworm.com/index.php?rid=5297412&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D21971847%26dopt%3DAbstract We describe experimental procedures for functional multineuron calcium imaging (fMCI) based on Nipkow-disk confocal microscopy, which enables us to monitor the activities of hundreds of neurons en masse at a cellular resolution at up to 2000 fps. Curr. Protoc. Neurosci. 57:2.14.1-2.14.10. © 2011 by John Wiley & Sons, Inc. PMID: 21971847 [PubMed - in process] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=5297412 Sat, 01 Oct 2011 04:00:00 +0100 5297412 http://www.medworm.com/index.php?rid=5297411&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D21971848%26dopt%3DAbstract Authors: Gray SJ, Choi VW, Asokan A, Haberman RA, McCown TJ, Samulski RJ Abstract Adeno-associated virus is a nonpathogenic human virus that has been developed into a gene-delivery vector due to its high efficiency of infection for many different cell types and its ability to persist and lead to long-term gene expression. This unit describes efficient methods to generate high-titer, research-grade, adenovirus-free recombinant single-stranded and self-complementary adeno-associated virus in various serotypes, along with methods to quantify the viral vectors. Two detailed methods are provided for viral vector delivery into the rodent brain and spinal cord, and for histological detection of transgene expression of GFP. Curr. Protoc. Neurosci. 57:4.17.1-4.17.30. © 2011 by John Wiley &... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=5297411 Sat, 01 Oct 2011 04:00:00 +0100 5297411 http://www.medworm.com/index.php?rid=5297410&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D21971849%26dopt%3DAbstract Authors: Ehrengruber MU, Schlesinger S, Lundstrom K Abstract Alphaviral vectors based on Semliki Forest virus and Sindbis virus infect many host cell types, causing rapid and high-level transgene expression. In the CNS, Semliki Forest virus and Sindbis virus exhibit an outstanding preference for neurons rather than glial cells, compared to other viruses. Generation of high-titer virus stocks is rapid (less than two days) and typically requires biosafety level 1 or 2 containment. Wild-type vectors are cytotoxic, permitting short-term transgene expression. However, mutant vectors with decreased cytotoxicity, to prolong host cell survival, have been developed. They also increase transgene expression and cellular co-infection, permitting heteromeric protein expression in individual cel... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=5297410 Sat, 01 Oct 2011 04:00:00 +0100 5297410 http://www.medworm.com/index.php?rid=5297409&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D21971850%26dopt%3DAbstract Authors: Jeon D, Shin HS Abstract Research on observed fear and its relation to human mental disorders has been hampered by the lack of a suitable animal model. The empathetic response, which is impaired in various mental disorders, requires the ability to recognize the emotions and feelings of others. Due to the lack of a robust behavioral assay system, studies of empathy in laboratory animals have been absent from the literature. This unit describes a protocol for assessing social observational fear learning as a precursor of empathy in the mouse. In this assay, the observer animal is conditioned for context-dependent fear by observing the behavior of the demonstrator animal receiving aversive stimuli. The magnitude of the fear response of the observer is positively influenced by... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=5297409 Sat, 01 Oct 2011 04:00:00 +0100 5297409 http://www.medworm.com/index.php?rid=5008854&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D21732311%26dopt%3DAbstract Authors: Gallagher SR Quantitation of nucleic acids is a fundamental tool in molecular biology that requires accuracy, reliability, and the use of increasingly smaller sample volumes. This unit describes the traditional absorbance measurement at 260 nm and three more sensitive fluorescence techniques employing Hoechst 33258, ethidium bromide, and PicoGreen. The range of the assays covers 25 pg/ml to 50 µg/ml. Absorbance at 260 nm has an effective range from 1 to 50 µg/ml; Hoechst 33258 from 0.01 to 15 µg/ml; ethidium bromide from 0.1 to 10 µg/ml; and PicoGreen from 25 to 1000 pg/ml. Curr. Protoc. Neurosci. 56:A.1K.1-A.1K.14. © 2011 by John Wiley & Sons, Inc. PMID: 21732311 [PubMed - in process] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=5008854 Thu, 30 Jun 2011 23:00:00 +0100 5008854 http://www.medworm.com/index.php?rid=5008853&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D21732312%26dopt%3DAbstract Authors: Smith CL This unit introduces the reader to the basic principles of confocal microscopy and the design and capabilities of current confocal microscopes. The advantages and disadvantages of confocal microscopy compared to other techniques for fluorescence imaging are described. There are also practical guidelines for sample preparation and optimization of imaging parameters, as well as examples of some of the applications of confocal microscopy. Curr. Protoc. Neurosci. 56:2.2.1-2.2.18. © 2011 by John Wiley & Sons, Inc. PMID: 21732312 [PubMed - in process] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=5008853 Thu, 30 Jun 2011 23:00:00 +0100 5008853 http://www.medworm.com/index.php?rid=5008852&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D21732313%26dopt%3DAbstract Authors: Ito K, Fujita K The neural crest is a unique structure in vertebrates. Neural crest cells play important roles in the formation of organs that characterize the vertebrate body plan. In this unit, we describe a primary culture method for mouse mesencephalic neural crest cells. The neural crest cells cultured by this method actively proliferate and differentiate into various cell types that originate from cranial neural crest cells, such as chondrocytes, neurons, and glia. Therefore, this primary culture method is useful for analyzing the development of mouse mesencephalic neural crest cells. Curr. Protoc. Neurosci. 56:3.23.1-3.23.8. © 2011 by John Wiley & Sons, Inc. PMID: 21732313 [PubMed - in process] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=5008852 Thu, 30 Jun 2011 23:00:00 +0100 5008852 http://www.medworm.com/index.php?rid=5008851&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D21732314%26dopt%3DAbstract Authors: Yang M, Silverman JL, Crawley JN Autism is diagnosed by three major symptom categories: unusual reciprocal social interactions, impaired communication, and repetitive behaviors with restricted interests. Direct social approach in mice has strong face validity to simple social approach behaviors in humans, which are frequently impaired in autism. This unit presents a basic protocol for a standardized, high-throughput social approach test for assaying mouse sociability. Our automated three-chambered social approach task quantifies direct social approach behaviors when a subject mouse is presented with the choice of spending time with either a novel mouse or a novel object. Sociability is defined as the subject mouse spending more time in the chamber containing the novel target m... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=5008851 Thu, 30 Jun 2011 23:00:00 +0100 5008851 http://www.medworm.com/index.php?rid=4704315&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D21462159%26dopt%3DAbstract Authors: Staffend NA, Meisel RL Identifying neuronal morphology is a key component in understanding neuronal function. Several techniques have been developed to address this issue, including Golgi staining, electroporation of fluorescent dyes, and transfection of fluorescent constructs. Ballistic delivery of transgenic constructs has been a successful means of rapidly transfecting a nonbiased population of cells within tissue or culture. Recently, this technique was modified for the ballistic delivery of dye-coated gold or tungsten particles, enabling a nonbiased, rapid fluorescent membrane labeling of individual neurons in both fixed and nonfixed tissue. This unit outlines a step-by-step protocol for the ballistic method of dye delivery ("DiOlistic" labeling) to fixed tissue, includin... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=4704315 Thu, 31 Mar 2011 23:00:00 +0100 4704315 http://www.medworm.com/index.php?rid=4704314&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D21462160%26dopt%3DAbstract Authors: Mortensen R Gene targeting by homologous recombination is a powerful and widely used technique for introduction of specific gene mutations (frequently a gene inactivation) in transgenic animals. The basic method detailed in this unit uses sequences homologous to the endogenous gene flanking the mutation. While methods using bacterial artificial chromosomes (BACs) and recombineering may be used, in most cases simpler bacterial plasmid clones with several kb of homology are sufficient. This protocol details the strategic factors in designing the constructs for selection and screening for homologous recombination. Curr. Protoc. Neurosci. 55:4.30.1-4.30.12. © 2011 by John Wiley & Sons, Inc. PMID: 21462160 [PubMed - in process] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=4704314 Thu, 31 Mar 2011 23:00:00 +0100 4704314 http://www.medworm.com/index.php?rid=4704313&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D21462161%26dopt%3DAbstract Authors: Golemis EA, Serebriiskii I, Finley RL, Kolonin Hunt By Interaction Mating MG, Gyuris J, Brent R The yeast two-hybrid method (or interaction trap) is a powerful technique for detecting protein interactions. The procedure is performed using transcriptional activation of a dual reporter system in yeast to identify interactions between a protein of interest (the bait protein) and the candidate proteins for interaction. The method can be used to screen a protein library for interactions with a bait protein or to test for association between proteins that are expected to interact based on prior evidence. Interaction mating facilitates the screening of a library with multiple bait proteins. Curr. Protoc. Neurosci. 55:4.4.1-4.4.35. © 2011 by John Wiley & Sons, Inc. PMID: 2146... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=4704313 Thu, 31 Mar 2011 23:00:00 +0100 4704313 http://www.medworm.com/index.php?rid=4704312&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D21462162%26dopt%3DAbstract Authors: Castagné V, Moser P, Roux S, Porsolt RD The development of antidepressants requires simple rodent behavioral tests for initial screening before undertaking more complex preclinical tests and clinical evaluation. Presented in the unit are two widely used screening tests used for antidepressants, the forced swim (also termed behavioral despair) test in the rat and mouse, and the tail suspension test in the mouse. These tests have good predictive validity and allow rapid and economical detection of substances with potential antidepressant-like activity. The behavioral despair and the tail suspension tests are based on the same principle: measurement of the duration of immobility when rodents are exposed to an inescapable situation. The majority of clinically used antidepressants... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=4704312 Thu, 31 Mar 2011 23:00:00 +0100 4704312 http://www.medworm.com/index.php?rid=4343301&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D21207365%26dopt%3DAbstract Authors: Apperson ML, Agius MA Multiple sclerosis (MS) is an immune-mediated disorder causing inflammation and demyelination in the central nervous system. As the onset of multiple sclerosis is at a young age, it is one of the leading neurological causes of disability. Disease activity and disability can be measured by neurological assessments and by magnetic resonance imaging. The development of standardized assessments has been a very important step in clinical research in MS. Clinical research in MS has led to a better understanding of the disease itself and has resulted in exciting new therapies. The protocols provided in this unit are four basic clinical and neuroimaging assessments commonly used as outcome measures in clinical research studies of MS subjects. These step-by-step i... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=4343301 Sat, 01 Jan 2011 00:00:00 +0100 4343301 http://www.medworm.com/index.php?rid=4343300&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D21207366%26dopt%3DAbstract Authors: Bortolotto ZA, Amici M, Anderson WW, Isaac JT, Collingridge GL Synaptic plasticity is the process by which the brain alters the strength of its synaptic connections, a fundamental function of the brain that enables individuals to learn from experience. The study of synaptic plasticity often involves the application of standard in vitro electrophysiological techniques to hippocampal slice preparations. This unit discusses many of the special considerations that are applicable for the optimal study of synaptic plasticity in this system. Most of these principles also apply to the study of synaptic plasticity in other brain slice preparations. Curr. Protoc. Neurosci. 54:6.13.1-6.13.26. © 2011 by John Wiley & Sons, Inc. PMID: 21207366 [PubMed - in process] (Source: Current... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=4343300 Sat, 01 Jan 2011 00:00:00 +0100 4343300 http://www.medworm.com/index.php?rid=4343297&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D21207367%26dopt%3DAbstract Authors: Russell VA Attention-deficit/hyperactivity disorder (ADHD) is a heterogeneous, highly heritable, behavioral disorder that affects ∼5% to 10% of children worldwide. Although animal models cannot truly reflect human psychiatric disorders, they can provide insight into the disorder that cannot be obtained from human studies because of the limitations of available techniques. Genetic models include the spontaneously hypertensive rat (SHR), the Naples High Excitability (NHE) rat, poor performers in the 5-choice serial reaction time (5-CSRT) task, the dopamine transporter (DAT) knock-out mouse, the SNAP-25 deficient mutant coloboma mouse, mice expressing a human mutant thyroid hormone receptor, a nicotinic receptor knock-out mouse, and a tachykinin-1 (NK1) receptor knock-out mouse... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=4343297 Sat, 01 Jan 2011 00:00:00 +0100 4343297 http://www.medworm.com/index.php?rid=4343295&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D21207368%26dopt%3DAbstract Authors: Stone EA, Lin Y This protocol describes a simplified method for inducing a chronic depression-like state in mice that is based on the repeated open-space forced swim method originally developed for rats. The method consists of mice swimming daily in lukewarm water in rat tub cages, for 15  min/day for 4 days, and thereafter once per week. This procedure produces a progressive decrease in distance swum and a concomitant increase in immobility (floating) in ∼70% of the mice, both of which persist unaltered for weeks. The model has predictive, face, and construct validity and has a number of advantages over previous methods in that it utilizes very mild stress, is short in duration, and is easily standardized. Moreover, since it utilizes a greater swimming area than the tradit... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=4343295 Sat, 01 Jan 2011 00:00:00 +0100 4343295 http://www.medworm.com/index.php?rid=4064521&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D20938922%26dopt%3DAbstract Authors: Grozdanov V, Müller A, Sengottuvel V, Leibinger M, Fischer D Retinal ganglion cells (RGCs) are central nervous system neurons with a very limited ability for axon regeneration. This unit details a cell culture technique, which can be used to functionally screen factors/compounds for their neuritogenic and neuroprotective effects on RGCs. In this protocol, the retina is isolated, digested in a papain solution, and after trituration, the RGCs are cultured. The neuritogenic effect of applied factors/compounds on RGCs in the medium is functionally determined by measuring the average neurite length of βIII-tubulin-positive RGCs in culture after 3 days. This protocol takes 3 to 7 days to perform depending on the application to complete, and is suitable to reliably test pharmacolog... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=4064521 Thu, 30 Sep 2010 23:00:00 +0100 4064521 http://www.medworm.com/index.php?rid=4064520&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D20938923%26dopt%3DAbstract Authors: Barde I, Salmon P, Trono D Lentiviral vectors have emerged over the last decade as powerful, reliable, and safe tools for stable gene transfer in a wide variety of mammalian cells. Unlike other vectors derived from oncoretroviruses, they allow for stable gene delivery into most nondividing primary cells, including neurons. This is why lentivectors (LVs) are becoming the most useful and promising tools in the field of neuroscience, not only for research, but also for future gene and cell therapy approaches. LVs derived from HIV-1 have gradually evolved to display many desirable features aimed at increasing both their safety and their versatility. These latest designs are reviewed in this unit. This unit also describes protocols for production and titration of LVs that can be im... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=4064520 Thu, 30 Sep 2010 23:00:00 +0100 4064520 http://www.medworm.com/index.php?rid=4064519&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D20938924%26dopt%3DAbstract Authors: O'Connor EC, Stephens DN, Crombag HS In appetitive Pavlovian associative learning, a stimulus (conditioned stimulus, CS) that has been associated with the delivery of a reinforcing event (unconditioned stimulus, US; e.g., food) can subsequently elicit or modulate goal-directed instrumental behaviors. For example, a Pavlovian CS can serve to reinforce (novel) instrumental behavior (conditioned reinforcement or CRf), or it can energize and potentiate ongoing instrumental responses when presented non-contingently (Pavlovian-instrumental transfer or PIT). Notably, these different effects of a Pavlovian CS on instrumental behavior are mediated by dissociable psychological and neurobiological mechanisms. Given the critical role that Pavlovian-instrumental interactions play in regula... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=4064519 Thu, 30 Sep 2010 23:00:00 +0100 4064519 http://www.medworm.com/index.php?rid=4064518&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D20938925%26dopt%3DAbstract Authors: Mathiasen JR, Dicamillo A Neuropsychiatric disorders encompass a broad patient population in a variety of disease states across all age groups and are often accompanied by deficits in short-term/working memory. However, most preclinical models that allow for an assessment of cognitive enhancement do not provide robust behavioral readouts with a level of throughput sufficient to support modern drug discovery efforts. The rat social recognition assay presented in this unit is one exception that has been increasingly employed to test new chemical entities for enhancing cognitive activity. The test is simple in design and takes advantage of the spontaneous behavior of rats to investigate conspecifics. The protocol in this unit is designed to evaluate the effects of a test substanc... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=4064518 Thu, 30 Sep 2010 23:00:00 +0100 4064518 http://www.medworm.com/index.php?rid=4064517&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D20938926%26dopt%3DAbstract Authors: Chan JS, Waldinger MD, Olivier B, Oosting RS This unit describes the testing of sexual behaviors of male Wistar rats. The described test enables the detection of stimulatory and inhibitory profiles of compounds. The test includes four training sessions to reach a stable sexual performance, followed by acute and/or chronic administration of drugs. The main quantifiable sexual behaviors are number of mounts (no vaginal penetration), intromissions (vaginal penetration), and ejaculations. By comparing the test compound to reference compound(s), sexual (side) effects can be determined. Curr. Protoc. Neurosci. 53:9.34.1-9.34.11. © 2010 by John Wiley & Sons, Inc. PMID: 20938926 [PubMed - in process] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=4064517 Thu, 30 Sep 2010 23:00:00 +0100 4064517 http://www.medworm.com/index.php?rid=3702107&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D20578033%26dopt%3DAbstract Authors: Light GA, Williams LE, Minow F, Sprock J, Rissling A, Sharp R, Swerdlow NR, Braff DL Understanding the basic neural processes that underlie complex higher-order cognitive operations and functional domains is a fundamental goal of cognitive neuroscience. Electroencephalography (EEG) is a non-invasive and relatively inexpensive method for assessing neurophysiological function that can be used to achieve this goal. EEG measures the electrical activity of large, synchronously firing populations of neurons in the brain with electrodes placed on the scalp. This unit outlines the basics of setting up an EEG experiment with human participants, including equipment, and a step-by-step guide to applying and preparing an electrode cap. Also included are support protocols for two event-rel... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=3702107 Sun, 27 Jun 2010 20:24:15 +0100 3702107 http://www.medworm.com/index.php?rid=3702106&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D20578034%26dopt%3DAbstract Authors: Kristian T This unit contains a protocol describing the isolation of brain mitochondria by using discontinuous Percoll gradient centrifugation. The Percoll density gradient centrifugation separates synaptosomes, myelin, and free nonsynaptic mitochondria released from cells during tissue homogenization into individual fractions. Mitochondria entrapped in synaptosomes (synaptic mitochondria) can be liberated using nitrogen cavitation and then further purified by Percoll gradient centrifugation. These methods yield mitochondria that exhibit good respiratory coupling and high respiratory rates. Curr. Protoc. Neurosci. 52:7.22.1-7.22.12. (c) 2010 by John Wiley & Sons, Inc. PMID: 20578034 [PubMed - in process] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=3702106 Sun, 27 Jun 2010 20:24:12 +0100 3702106 http://www.medworm.com/index.php?rid=3702105&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D20578035%26dopt%3DAbstract Authors: Motulsky HJ, Neubig RR Measuring the rate and extent of radioligand binding provides information on the number of binding sites, and their affinity and accessibility of these binding sites for various drugs. This unit explains how to design and analyze such experiments. Curr. Protoc. Neurosci. 52:7.5.1-7.5.65. (c) 2010 by John Wiley & Sons, Inc. PMID: 20578035 [PubMed - in process] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=3702105 Sun, 27 Jun 2010 20:24:09 +0100 3702105 http://www.medworm.com/index.php?rid=3702104&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D20578036%26dopt%3DAbstract Authors: Coccurello R, Moles A In comparison with conventional, first-generation antipsychotics (e.g., haloperidol), the administration of atypical antipsychotics (AAPs) has been associated with a higher risk of metabolic derangements, including body weight increase, dysregulation of glucose homeostasis, fat accumulation, and even liability to develop type II diabetes. Since this is a serious clinical problem that may be further exacerbated in overweight schizophrenics, establishing animal models of AAP-induced adverse effects may contribute to clarifying the mechanisms underlying these effects. Here we present three basic protocols by which this problem has been modeled. The three protocols differ in many aspects (routes of administration, extent of the chronic treatment, diets, and d... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=3702104 Sun, 27 Jun 2010 20:24:06 +0100 3702104 http://www.medworm.com/index.php?rid=3450159&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D20373505%26dopt%3DAbstract Authors: Driver EC, Kelley MW The sensory epithelium of the mammalian inner ear, also referred to as the organ of Corti, is a remarkable structure comprised of highly ordered rows of mechanosensory hair cells and non-sensory supporting cells located within the coiled cochlea. This unit describes an in vitro explant culture technique that can be coupled with gene transfer via electroporation to study the effects of altering gene expression during development of the organ of Corti. While the protocol is largely focused on embryonic cochlea, the same basic protocol can be used on cochleae from mice as old as P5. PMID: 20373505 [PubMed - in process] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=3450159 Wed, 31 Mar 2010 23:00:00 +0100 3450159 http://www.medworm.com/index.php?rid=3450158&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D20373506%26dopt%3DAbstract Authors: Bradbury AR Phage display has been extensively used to study protein-protein interactions, receptor- and antibody-binding sites, and immune responses, to modify protein properties, and to select antibodies against a wide range of different antigens. In the format most often used, a polypeptide is displayed on the surface of a filamentous phage by genetic fusion to one of the coat proteins, creating a chimeric coat protein, and coupling phenotype (the protein) to genotype (the gene within). As the gene encoding the chimeric coat protein is packaged within the phage, selection of the phage on the basis of the binding properties of the polypeptide displayed on the surface simultaneously results in the isolation of the gene encoding the polypeptide. This unit describes the backgro... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=3450158 Wed, 31 Mar 2010 23:00:00 +0100 3450158 http://www.medworm.com/index.php?rid=3450157&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D20373507%26dopt%3DAbstract Authors: Vidi PA, Przybyla JA, Hu CD, Watts VJ Members of the G protein-coupled receptor (GPCR) superfamily have been shown to homo- and hetero-oligomerize both in vitro and in vivo. Although the functional and pharmacological significance of GPCR oligomerization is far from being completely understood, evidence suggests that, depending on the receptor, oligomerization may influence ligand binding, G protein coupling, and receptor targeting. Bimolecular fluorescence complementation (BiFC) is a technique based on the complementation of fragments from fluorescent proteins that allows the measurement and visualization of protein interactions in living cells. It can be extended to the simultaneous detection of distinct protein-protein interactions using a multicolor setup. This unit descri... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=3450157 Wed, 31 Mar 2010 23:00:00 +0100 3450157 http://www.medworm.com/index.php?rid=3450156&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D20373508%26dopt%3DAbstract Authors: June HL, Gilpin NW Operant self-administration procedures are used to assess the neural basis of ethanol-seeking behavior under a wide range of experimental conditions. In general, rats do not spontaneously self-administer ethanol in pharmacologically meaningful amounts. This unit provides a step-by-step guide for training rats to self-administer quantities of ethanol that produce moderate to high blood-alcohol content. Different protocols are used for rats that are genetically heterogeneous versus rats that are selectively bred for high alcohol preference. Also, these protocols have different sets of advantages and disadvantages in terms of the ability to control for caloric intake and taste of solutions in operant testing. Basic self-administration protocols can also be alte... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=3450156 Wed, 31 Mar 2010 23:00:00 +0100 3450156 http://www.medworm.com/index.php?rid=3172666&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D20066654%26dopt%3DAbstract Authors: Xu Q, Anderson SA As the brain develops, progenitor cells acquire the features of specific neuronal or glial subtypes through dynamic expression of the fate-determining signaling molecules and their targeting transcription factors. An effective and versatile approach for tracing lineage of progenitors into adult cell types is to target the promoter of an interested gene with Cre (a phage DNA recombinase) to achieve simultaneous activation during neurogenesis. The bacterial artificial chromosome (BAC) is an efficient Cre carrier. Not only the targeted gene remains diploidy in BAC-Cre transgenic mice, but also the large portions of the gene's regulatory elements to be incorporated in the BAC allow Cre to sufficiently and reliably reproduce the endogenous gene expression pattern.... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=3172666 Fri, 01 Jan 2010 00:00:00 +0100 3172666 http://www.medworm.com/index.php?rid=3172665&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D20066655%26dopt%3DAbstract Authors: Combs CA The field of fluorescence microscopy is rapidly growing, providing ever increasing imaging capabilities for cell and neurobiologists. Over the last decade, many new technologies and techniques have been developed which allow for deeper, faster, or higher resolution imaging. For the non-expert microscopist, it can be difficult to match the best imaging technique to the biological question to be examined. Picking the right technique requires a basic understanding of the underlying imaging physics for each technique, as well as an informed comparison and balancing of competing imaging properties in the context of the sample to be imaged. This unit provides concise descriptions of a range of commercially available imaging techniques and provides a tabular guide to choosin... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=3172665 Fri, 01 Jan 2010 00:00:00 +0100 3172665 http://www.medworm.com/index.php?rid=3172664&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D20066656%26dopt%3DAbstract Authors: Delaney KR Dextran-conjugated Ca(2+) indicators are retained well in neurons for many days following loading in intact or semi-intact brain tissue. Methods for loading neurons, as well as discussion of the unique properties of dextran-conjugated dyes which need to be considered for their use, are presented. PMID: 20066656 [PubMed - in process] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=3172664 Fri, 01 Jan 2010 00:00:00 +0100 3172664 http://www.medworm.com/index.php?rid=3172663&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D20066657%26dopt%3DAbstract Authors: Puntel M, Kroeger KM, Sanderson NS, Thomas CE, Castro MG, Lowenstein PR Viral vector-mediated gene delivery is an attractive procedure for introducing genes into the brain, both for purposes of basic neuroscience research and to develop gene therapy for neurological diseases. Replication-defective adenoviruses possess many features which make them ideal vectors for this purpose-efficiently transducing terminally differentiated cells such as neurons and glial cells, resulting in high levels of transgene expression in vivo. Also, in the absence of anti-adenovirus immunity, these vectors can sustain very long-term transgene expression within the brain parenchyma. This unit provides protocols for the stereotactic injection of adenoviral vectors into the brain, followed by protocol... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=3172663 Fri, 01 Jan 2010 00:00:00 +0100 3172663 http://www.medworm.com/index.php?rid=3172662&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D20066658%26dopt%3DAbstract Authors: Pei Y, Dong S, Roth BL G protein-coupled receptors (GPCRs) and their signal transductions are important for both physiological and pathological processes in neuron systems. Neuronal GPCRs activated by synthetic ligands have been created by designed mutagenesis for studying their functions and signal pathways. However, these engineered GPCRs have problems, such as their high constitutive activity. To overcome this drawback, a new generation of receptors termed designer receptors exclusively activated by designer drugs (DREADDs), have been designed. DREADDs are exclusively activated by synthetic ligands, but are insensitive to their endogenous ligand and have no constitutive activity, which provides the ability to selectively modulate signal transduction of certain GPCRs in vitr... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=3172662 Fri, 01 Jan 2010 00:00:00 +0100 3172662 http://www.medworm.com/index.php?rid=2866132&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19802812%26dopt%3DAbstract Authors: Perlmutter JS Parkinson disease (PD) is a progressive neurologic condition that causes motor and nonmotor manifestations. Treatment provides symptomatic benefit but no current treatment has been proven to slow disease progression. Research studies of PD require a means of rating the severity of disease by measurement of motor manifestations, assessment of ability to perform daily functional activities, and symptomatic response to medication. The most common rating scales are the Unified Parkinson Disease Rating Scale (UPDRS), Hoehn and Yahr staging, and the Schwab and England rating of activities of daily living. Each of these rating scales are described, including detailed instructions on how to implement these ratings. Although these are the most widely applied rating scales... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2866132 Wed, 30 Sep 2009 23:00:00 +0100 2866132 http://www.medworm.com/index.php?rid=2866131&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19802813%26dopt%3DAbstract Authors: Campbell IG The electroencephalogram (EEG) is the most common tool used in sleep research. This unit describes the methods for recording and analyzing the EEG. Detailed protocols describe recorder calibration, electrode application, EEG recording, and computer EEG analysis with power spectral analysis. Computer digitization of an analog EEG signal is discussed, along with EEG filtering and the parameters of fast Fourier transform (FFT) power spectral analysis. Sample data are provided for a typical night's analysis of EEG during NREM (non-REM) and REM sleep. PMID: 19802813 [PubMed - in process] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2866131 Wed, 30 Sep 2009 23:00:00 +0100 2866131 http://www.medworm.com/index.php?rid=2866130&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19802814%26dopt%3DAbstract Authors: Baldwin S, Farias ST Alzheimer's disease (AD) is the most common form of dementia in older adults. It represents a significant public health concern because of its associated personal, social, and economic burden. As such, AD is the focus of considerable research worldwide. This unit reviews the major cognitive and behavioral impairments associated with AD, and the practical application of current neuropsychological procedures used to assess these deficits. PMID: 19802814 [PubMed - in process] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2866130 Wed, 30 Sep 2009 23:00:00 +0100 2866130 http://www.medworm.com/index.php?rid=2866129&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19802815%26dopt%3DAbstract Authors: Fields DR, Shneider N, Mentis GZ, O'Donovan MJ This unit describes methods for loading ion- and voltage-sensitive dyes into neurons, with a particular focus on the spinal cord as a model system. In addition, we describe the use of these dyes to visualize neural activity. Although the protocols described here concern spinal networks in culture or an intact in vitro preparation, they can be, and have been, widely used in other parts of the nervous system. Curr. Protoc. Neurosci. 49:2.3.1-2.3.16. (c) 2009 by John Wiley & Sons, Inc. PMID: 19802815 [PubMed - in process] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2866129 Wed, 30 Sep 2009 23:00:00 +0100 2866129 http://www.medworm.com/index.php?rid=2866128&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19802816%26dopt%3DAbstract Authors: Martín Del Campo C, Velázquez JL, Freire MA This unit describes the materials, methods, and analytical techniques available for the study of electrical activity of neural tissue in rodents in both homeostatic and disease states, with emphasis on epileptogenesis. A table containing a list of suppliers of relevant materials and equipment is also provided. Curr. Protoc. Neurosci. 49:6.24.1-6.24.24. (c) 2009 by John Wiley & Sons, Inc. PMID: 19802816 [PubMed - in process] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2866128 Wed, 30 Sep 2009 23:00:00 +0100 2866128 http://www.medworm.com/index.php?rid=2866127&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19802817%26dopt%3DAbstract Authors: Gourley SL, Taylor JR Alterations in multiple biological functions, such as transcription factor activity, are implicated in the neurobiology of depression, based primarily on the characterization of antidepressant efficacy in naïve rodents rather than on models that capture the protracted feelings of anhedonia and helplessness that typify depression. This unit presents rat and mouse models of depression that involve chronic oral exposure to the stress-associated adrenal hormone, corticosterone (CORT), resulting in anhedonic- and helplessness-like behaviors that are persistent yet reversible by chronic antidepressant treatment. Prior CORT exposure also chronically influences molecular targets hypothesized to contribute to negative mood. One example is phosphorylation of c... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2866127 Wed, 30 Sep 2009 23:00:00 +0100 2866127 http://www.medworm.com/index.php?rid=2573540&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19575469%26dopt%3DAbstract Authors: Caligioni CS The short reproductive cycle length observed in rodents, called the estrous cycle, makes them an ideal animal model for investigation of changes that occur during the reproductive cycle. Most of the data in the literature about the estrous cycle is obtained from rats because they are easily manipulated and they exhibit a clear and well-defined estrous cycle. However, the increased number of experiments using knockout mice requires identification of their estrous cycle as well, since (in)fertility issues may arise. In mice, like rats, the identification of the stage of estrous cycle is based on the proportion of cell types observed in the vaginal secretion. The aim of this unit is to provide guidelines for quickly and accurately determining estrous cycle phases in ... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2573540 Tue, 30 Jun 2009 23:00:00 +0100 2573540 http://www.medworm.com/index.php?rid=2573539&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19575470%26dopt%3DAbstract Authors: Yung YC, Kennedy G, Chun J Programmed cell death is a fundamental process for the development and somatic maintenance of organisms. This unit describes methods for visualizing both dying cells in situ and for detection of nucleosomal ladders. A description of various current detection strategies is provided, as well as support protocols for preparing positive and negative controls and for preparing genomic DNA. Curr. Protoc. Neurosci. 48:3.8.1-3.8.24. (c) 2009 by John Wiley & Sons, Inc. PMID: 19575470 [PubMed - as supplied by publisher] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2573539 Tue, 30 Jun 2009 23:00:00 +0100 2573539 http://www.medworm.com/index.php?rid=2573538&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19575471%26dopt%3DAbstract Authors: Cooper HM, Paterson Y Much of modern biology and biochemistry relies on the availability of highly specific antibodies for use in such ubiquitous techniques as immunohistochemistry, ELISAs, immunoprecipitation, and immunoblotting. Thus, the generation of large quantities of specific antibodies directed against proteins or peptides of interest is essential to the success of both basic and applied research programs. In addition, with the advent of antibody-based proteomic strategies for profiling protein expression and post-translational modification, a requirement for timely production of specific antibodies has been exemplified. Polyclonal antibodies derived from animals immunized with purified proteins or peptides are particularly valuable for use in the laboratory. This unit... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2573538 Tue, 30 Jun 2009 23:00:00 +0100 2573538 http://www.medworm.com/index.php?rid=2573537&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19575472%26dopt%3DAbstract Authors: Hall ED, Bosken JM One of the most completely validated processes involved in secondary tissue damage following acute brain or spinal cord injury and in many chronic neurodegenerative diseases has to do with the pathological formation of reactive oxygen species (ROS) and reactive nitrogen species (RNS). These are generated by multiple mechanisms and give rise to highly reactive oxygen radicals that can damage neuronal, glial, and microvascular elements. Particular interest has centered upon oxygen radical-induced, iron-catalyzed lipid peroxidation (LP) as the principal mechanism of neuronal injury associated with oxygen radicals. Thus, there has been a growing interest in monitoring increased oxygen radical levels as an index of oxidative stress, as well as measuring markers o... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2573537 Tue, 30 Jun 2009 23:00:00 +0100 2573537 http://www.medworm.com/index.php?rid=2573536&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19575473%26dopt%3DAbstract Authors: Zapata A, Chefer VI, Shippenberg TS, Denoroy L Sensitive analytical methods are needed for the separation and quantification of neurotransmitters obtained in microdialysate studies. This unit describes methods that permit quantification of nanomolar concentrations of monoamines and their metabolites (high-pressure liquid chromatography electrochemical detection), acetylcholine (HPLC-coupled to an enzyme reactor), and amino acids (HPLC-fluorescence detection; capillary electrophoresis with laser-induced fluorescence detection).Curr. Protoc. Neurosci. 48:7.4.1-7.4.30. (c) 2009 by John Wiley & Sons, Inc. PMID: 19575473 [PubMed - as supplied by publisher] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2573536 Tue, 30 Jun 2009 23:00:00 +0100 2573536 http://www.medworm.com/index.php?rid=2573535&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19575474%26dopt%3DAbstract Authors: Yang M, Crawley JN This unit presents two basic protocols that offer rapid assessments of anosmia (the absence of a sense of smell) in mice. The buried food test is used to check for the ability to smell volatile odors. The olfactory habituation/dishabituation test is used to test whether the animal can detect and differentiate different odors, including both nonsocial and social odors. A non-contact method of odor presentation, along with a general method for collecting urine samples, is given as an alternate protocol. The tests described in this unit only require simple equipment and can be adopted readily by most laboratories.Curr. Protoc. Neurosci. 48:8.24.1-8.24.12. (c) 2009 by John Wiley & Sons, Inc. PMID: 19575474 [PubMed - as supplied by publisher] (Source: Cur... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2573535 Tue, 30 Jun 2009 23:00:00 +0100 2573535 http://www.medworm.com/index.php?rid=2525277&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19340810%26dopt%3DAbstract Authors: Mansouri A, Fukumitsu H, Schindehuette J, Krieglstein K Mouse embryonic stem (ES) cells are derived from mouse blastocyst and are able to generate all embryonic tissues in vitro. This propensity of ES cells has acquired considerable attention in recent years due to the promising potential for future cell replacement-based therapies. Therefore, it is of fundamental interest to establish protocols that allow the differentiation of ES cells into specific cell types. In recent years, several such differentiation procedures have been described for mouse and human embryonic stem cells. This unit describes a simple procedure that promotes the neuronal differentiation of mouse embryonic stem cells and yields a high proportion of midbrain dopaminergic neurons. Furthermore, this procedu... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2525277 Tue, 31 Mar 2009 23:00:00 +0100 2525277 http://www.medworm.com/index.php?rid=2525273&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19340811%26dopt%3DAbstract Authors: Zeitelhofer M, Vessey JP, Thomas S, Kiebler M, Dahm R Despite the development of various transfection methods, the transfection of post-mitotic cells, including neurons, poses a challenging task. Nucleofection, a specialized form of electroporation described in this unit, achieves high transfection efficiencies in primary mammalian neurons, such as hippocampal neurons, while simultaneously maintaining high cell viability. Therefore, it allows for biochemical analyses that rely on large numbers of transfected cells. The recently developed 96-well shuttle system described in this unit further permits the transfection of up to 96 different constructs in a single experiment. This opens up the possibility for large-scale experiments in primary neurons, such as shRNA-mediated knock-... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2525273 Tue, 31 Mar 2009 23:00:00 +0100 2525273 http://www.medworm.com/index.php?rid=2525270&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19340812%26dopt%3DAbstract Authors: Chefer VI, Thompson AC, Zapata A, Shippenberg TS The technique of microdialysis enables sampling and collecting of small-molecular-weight substances from the interstitial space. It is a widely used method in neuroscience and is one of the few techniques available that permits quantification of neurotransmitters, peptides, and hormones in the behaving animal. More recently, it has been used in tissue preparations for quantification of neurotransmitter release. This unit provides a brief review of the history of microdialysis and its general application in the neurosciences. The authors review the theoretical principles underlying the microdialysis process, methods available for estimating extracellular concentration from dialysis samples (i.e., relative recovery), the various f... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2525270 Tue, 31 Mar 2009 23:00:00 +0100 2525270 http://www.medworm.com/index.php?rid=2525267&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19340813%26dopt%3DAbstract Authors: Zapata A, Chefer VI, Shippenberg TS Microdialysis is an in vivo sampling technique that permits the quantification of various substances (e.g., neurotransmitters, peptides, electrolytes) in blood and tissue. It is also used to infuse substances into the brain and spinal cord. This unit describes methods for the construction and stereotaxic implantation of microdialysis probes into discrete brain regions of the rat and mouse. Procedures for the conduct of conventional and quantitative microdialysis experiments in the awake and anesthetized rodent are also provided. PMID: 19340813 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2525267 Tue, 31 Mar 2009 23:00:00 +0100 2525267 http://www.medworm.com/index.php?rid=2525265&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19340814%26dopt%3DAbstract Authors: Chang CH, Knapska E, Orsini CA, Rabinak CA, Zimmerman JM, Maren S Pavlovian conditioning paradigms have become important model systems for understanding the neuroscience of behavior. In particular, studies of the extinction of Pavlovian fear responses are yielding important information about the neural substrates of anxiety disorders, such as phobias and post-traumatic stress disorder (PTSD) in humans. These studies are germane to understanding the neural mechanisms underlying behavioral interventions that suppress fear, including exposure therapy in anxiety disorders. This unit describes detailed behavioral protocols for examining the nature and properties of fear extinction in laboratory rodents. PMID: 19340814 [PubMed - indexed for MEDLINE] (Source: Current Protocols in... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2525265 Tue, 31 Mar 2009 23:00:00 +0100 2525265 http://www.medworm.com/index.php?rid=2175341&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19170022%26dopt%3DAbstract Authors: Kostrzewa RM In the 1950s, the discovery of anti-nerve growth factor, an immunotoxin stunting sympathetic neural development, signaled the advent of neurotoxins as research modalities. Other selective neurotoxins were discovered in rapid succession. In the 1960s, 6-hydroxydopamine and 6-hydroxydopa were shown to destroy noradrenergic and dopaminergic nerves. Excitotoxins (glutamate, aspartate, and analogs) were discovered in the 1970s. DSP-4 [N-(2-chloroethyl)-N-ethyl-2-bromobenzylamine] proved to be selective for noradrenergic destruction, while 5,6- and 5,7-dihydroxytryptamines were relatively selective for serotonin neurons. Additional neurotoxins were discovered, but it was MPTP (1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine) that predominated neurotoxicity research in the ... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2175341 Thu, 01 Jan 2009 05:00:00 +0100 2175341 http://www.medworm.com/index.php?rid=2175340&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19170023%26dopt%3DAbstract Authors: Free RB, Hazelwood LA, Sibley DR Proteomics has evolved from genomic science due to the convergence of advances in protein chemistry, separations, mass spectroscopy, and peptide and protein databases. Where identifying protein-protein interactions was once limited to yeast two-hybrid analyses or empirical data, protein-protein interactions can now be examined in both cells and native tissues by precipitation of the protein complex of interest. Coupling this field to receptor pharmacology has recently allowed for the identification of proteins that differentially and selectively interact with receptors and are integral to their biological effects. It is becoming increasingly apparent that receptors in neurons do not exist as singular independent units, but rather are part of la... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2175340 Thu, 01 Jan 2009 05:00:00 +0100 2175340 http://www.medworm.com/index.php?rid=2175339&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D19170024%26dopt%3DAbstract Authors: Schneider T, Popik P Premenstrual dysphoric disorder (PMDD) is characterized by the recurrence of a cluster of physical and negative mood symptoms, especially irritability, appearing when estrogen and progesterone levels decrease during the late luteal phase of the menstrual cycle. This unit describes a new animal model of PMDD that shows differentiation between female rats expressing and not expressing ovarian cycle-dependent irritability measured by the behavior of burying harmless objects. Burying behavior is enhanced in a subgroup of female rats at metestrus and decreased at the proestrus phase of the estrous cycle. Increased marble burying at metestrus does not habituate and can be reversed by acute treatment with several antidepressants, which mimic the pharmacology of P... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=2175339 Thu, 01 Jan 2009 05:00:00 +0100 2175339 http://www.medworm.com/index.php?rid=1926951&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18633997%26dopt%3DAbstract Authors: Fasano C, Thibault D, Trudeau LE This unit presents a protocol for primary culture of postnatal mesencephalic dopamine neurons grown on an astrocyte monolayer, which can be used to investigate cellular and molecular mechanisms regulating dopamine neuron function. Using this in vitro approach, dopamine neurons survive for an extended period of time and establish functional axon terminals and dendrites that display properties similar to those observed in vivo and in brain slices. An alternate protocol is provided for a microculture system in which astrocytes are grown on a spatially limited surface where single or small groups of dopamine neurons develop. Under such conditions, isolated neurons establish synaptic contacts, or autapses, onto their own somatodendritic compartment,... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926951 Tue, 01 Jul 2008 04:00:00 +0100 1926951 http://www.medworm.com/index.php?rid=1926950&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18633998%26dopt%3DAbstract Authors: Morozov A Recently developed techniques for spatially and temporally controlled genetic manipulations based on regulated homologous recombination and/or transcription are extensively used in brain research. In addition to being important for testing the role of specific proteins in the central nervous system, these techniques allow analysis of brain functions at the neuronal circuit level. This overview discusses principles of conditional inactivation and expression of genes, and their specific applications to studies of the mammalian brain. PMID: 18633998 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926950 Tue, 01 Jul 2008 04:00:00 +0100 1926950 http://www.medworm.com/index.php?rid=1926949&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18633999%26dopt%3DAbstract Authors: Aras MA, Hartnett KA, Aizenman E This unit contains five protocols for assaying cell viability in vitro using primary neuronal cultures, including a novel method for use with transfected neurons. Three of the assays are based on the principle that cell death cascades alter membrane permeability. The lactate dehydrogenase (LDH) release assay measures the amount of the cytoplasmic enzyme released into the bathing medium, while the trypan blue and propidium iodide assays measure the ability of cells to exclude dye from their cytoplasm. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay measures the mitochondrial activity of viable cells by quantifying the conversion of the tetrazolium salt to its formazan product. Finally, the fifth assay details the mea... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926949 Tue, 01 Jul 2008 04:00:00 +0100 1926949 http://www.medworm.com/index.php?rid=1926948&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18634000%26dopt%3DAbstract Authors: Faingold CL Binge drinking of alcohol is an important public health issue, and experimental studies are needed to understand the pathophysiological mechanisms of this problem and develop improved approaches to treatment. This unit presents a validated and widely used method to model binge alcohol drinking in rats. It consists of three daily intragastric administrations of ethanol to rats for 4 days. Ethanol is initially administered at 5 g/kg, and then each subsequent dose is determined based on the degree of intoxication the rat exhibits prior to each dose. The behavior of the animal is graded based on a well-described scale. After the fourth day, various aspects of the ethanol withdrawal syndrome can be observed over a predictable time course and additional protocols can be ... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926948 Tue, 01 Jul 2008 04:00:00 +0100 1926948 http://www.medworm.com/index.php?rid=1926947&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18634001%26dopt%3DAbstract Authors: Gilpin NW, Richardson HN, Cole M, Koob GF Alcohol dependence constitutes a neuroadaptive state critical for understanding alcoholism, and various methods have been utilized to induce alcohol dependence in animals, one of which is alcohol vapor exposure. Alcohol vapor inhalation provides certain advantages over other chronic alcohol exposure procedures that share the ultimate goal of producing alcohol dependence in rats. Chronic alcohol vapor inhalation allows the experimenter to control the dose, duration, and pattern of alcohol exposure. Also, this procedure facilitates testing of somatic and motivational aspects of alcohol dependence. Chronic exposure to alcohol vapor produces increases in alcohol-drinking behavior, increases in anxiety-like behavior, and reward deficits in ... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926947 Tue, 01 Jul 2008 04:00:00 +0100 1926947 http://www.medworm.com/index.php?rid=1926956&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428673%26dopt%3DAbstract Authors: Schofield BR The growth of fluorescence imaging technology and the development of sensitive fluorescent retrograde tracers has provided many new approaches for analyzing neuronal circuits. Fluorescent markers provide unparalleled opportunity for combining axonal tract tracing with techniques such as immunohistochemistry or physiological recording. This unit describes the use of six different fluorescent tracers: Fast Blue, fluorescein dextran, FluoroGold, FluoroRuby, red beads, and green beads. Guidance is provided on how to choose a tracer for a particular experiment, and three methods are described for injecting the tracers, including pressure injection through a microsyringe or a micropipet, and iontophoretic injection through a micropipet. Criteria for selecting the most a... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926956 Tue, 01 Apr 2008 04:00:00 +0100 1926956 http://www.medworm.com/index.php?rid=1926955&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428674%26dopt%3DAbstract Authors: Morgan JL, Wong RO Neuronal cell labeling is fundamental to investigations of the nervous system. Labeling of cells in live or fixed tissue with dyes or ion indicators using ballistic approaches has recently been developed for the study of neuronal architecture and function. In this approach, dye-coated particles are propelled into cells by a pulse of pressurized helium. This unit provides step-by-step protocols for coating tungsten particles with fluorescent or indicator dyes and for delivering these particles into cells and tissue. The major advantage of the ballistic method of dye delivery is that large populations of neurons can be rapidly labeled within a piece of live or fixed tissue. Advantages and limitations of the approach are discussed and technical advice is provid... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926955 Tue, 01 Apr 2008 04:00:00 +0100 1926955 http://www.medworm.com/index.php?rid=1926954&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428675%26dopt%3DAbstract Authors: Cousin MA The fluorescent dye FM1-43 and its derivatives can be used to monitor the physiology of synaptic vesicle turnover in central nerve terminals. They do so by their ability to reversibly partition into membranes, a process that results in a huge increase in fluorescence in comparison to their quantum yield in solution. This unit provides protocols for quantifying total synaptic vesicle turnover, the kinetics and extent of synaptic vesicle exocytosis, and the kinetics and mode of synaptic vesicle endocytosis. Descriptions of other ways these protocols have been used to derive information about the life cycle of the synaptic vesicle are also provided. PMID: 18428675 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926954 Tue, 01 Apr 2008 04:00:00 +0100 1926954 http://www.medworm.com/index.php?rid=1926953&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428676%26dopt%3DAbstract Authors: Moore SW, Kennedy TE Studies of spinal commissural neurons have provided substantial insight into the mechanisms that regulate axon guidance. Explants of embryonic spinal cords and isolated spinal commissural neurons have been important experimental tools for the identification and characterization of several guidance cues, including netrins, semaphorins, slits, sonic hedgehog, BMPs, and wnts. In this unit, protocols are provided for establishing these explant assays to assess the outgrowth and turning capacity of commissural axons. In addition, methods are included for preparing cultures highly enriched with embryonic commissural neurons, which have been used to probe the biochemical signaling mechanisms regulating axon guidance. PMID: 18428676 [PubMed - indexed for MEDLI... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926953 Tue, 01 Apr 2008 04:00:00 +0100 1926953 http://www.medworm.com/index.php?rid=1926952&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428677%26dopt%3DAbstract Authors: Platt B, Beyer CE, Schechter LE, Rosenzweig-Lipson S Obsessive-compulsive disorder (OCD) is difficult to model in animals due to the involvement of both mental (obsessions) and physical (compulsions) symptoms. Due to limitations of using animals to evaluate obsessions, OCD models are limited to evaluation of the compulsive and repetitive behaviors of animals. Of these, models of adjunctive behaviors offer the most value in regard to predicting efficacy of anti-OCD drugs in the clinic. Adjunctive behaviors are those that are maintained indirectly by the variables that control another behavior, rather than directly by their own typical controlling variables. Schedule-induced polydipsia (SIP) is an adjunctive model in which rats exhibit exaggerated drinking behavior (polydipsia) ... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926952 Tue, 01 Apr 2008 04:00:00 +0100 1926952 http://www.medworm.com/index.php?rid=1926960&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428669%26dopt%3DAbstract Authors: Davis JA Many animal models used in neuroscience research must be surgically created and/or anesthetized for imaging studies. The purpose of this unit is to review the advantages and disadvantages of various anesthetic and analgesic agents in rodents; to discuss state-of-the-art methods for monitoring anesthesia; and to provide tips for troubleshooting problems with anesthesia. PMID: 18428669 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926960 Tue, 01 Jan 2008 05:00:00 +0100 1926960 http://www.medworm.com/index.php?rid=1926959&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428670%26dopt%3DAbstract Authors: Hallett PJ, Collins TL, Standaert DG, Dunah AW An important tool for studying the regulation of synapses is a rapid and reliable means of separating synaptic and intracellular proteins. This unit presents a technique for analysis of brain tissue which relies on differential centrifugation to separate proteins present at synaptic sites from those found in intracellular cytoplasmic and vesicular pools. The method is efficient in that only small amounts of tissue, such as might be obtained from a small region of a rodent brain, are required. It is reproducible and, in conjunction with immunoblot or immunoprecipitation techniques, can produce reliable quantitative data. The protocol will be of interest to those conducting a variety of different studies related to the localization ... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926959 Tue, 01 Jan 2008 05:00:00 +0100 1926959 http://www.medworm.com/index.php?rid=1926958&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428671%26dopt%3DAbstract Authors: Lee JP, McKercher S, Muller FJ, Snyder EY Neural stem cells (NSCs) are the most primordial, least committed cells of the nervous system, and transplantation of these multipotent cells holds the promise of regenerative therapy for many central nervous system (CNS) diseases. This unit describes methods for NSC transplantation into neonatal mouse pups, embryonic mouse brain, and adult mouse brain. A description of options for detection of labeled donor cells in engrafted mouse brain is provided along with an example protocol for detecting lacZ-expressing cells in situ. Also included is a protocol for preparing NSCs for transplantation. PMID: 18428671 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926958 Tue, 01 Jan 2008 05:00:00 +0100 1926958 http://www.medworm.com/index.php?rid=1926957&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428672%26dopt%3DAbstract Authors: Crabbe JC, Cameron AJ, Munn E, Bunning M, Wahlsten D There are many behavioral assays to assess sensitivity to ethanol intoxication in mice. Most are simple to implement, and are sensitive to a particular dose range of ethanol. Most reflect genetic influences, and each test appears to reflect the contribution of a relatively distinct collection of genes. This genetic heterogeneity implies that no single test can claim to capture the construct "ethanol intoxication" completely. Depending on the test, and when measurements are made, acute functional tolerance to even a single dose of ethanol must be considered as a contributing factor. Whether or not a test is conducted in naïve mice or as part of a test battery can influence sensitivity, and do so in a strain-dependent man... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926957 Tue, 01 Jan 2008 05:00:00 +0100 1926957 http://www.medworm.com/index.php?rid=1926973&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428656%26dopt%3DAbstract Authors: Ehrengruber MU, Lundstrom K Alphaviral vectors based on Semliki Forest virus and Sindbis virus infect many host cell types, causing rapid and high-level transgene expression. Compared to other viruses used to infect CNS cell and tissue preparations, Semliki Forest virus and Sindbis virus exhibit an outstanding preference for neurons rather than glial cells. High-titer vector generation typically requires biosafety level 1 or 2 containment and occurs in less than 2 days. Wild-type vectors are cytotoxic, permitting short-term transgene expression. However, mutant vectors with decreased cytotoxicity, to prolong host cell survival, have been developed. They also increase transgene expression and cellular coinfection, permitting heteromeric protein expression in individual cells. O... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926973 Mon, 01 Oct 2007 04:00:00 +0100 1926973 http://www.medworm.com/index.php?rid=1926970&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428659%26dopt%3DAbstract Authors: Grefen C, Lalonde S, Obrdlik P Protein-protein interactions play a fundamental role in the regulation of almost all cellular processes. Thus, the identification of interacting proteins can help to elucidate their function. The mating-based split-ubiquitin system (mbSUS) uses yeast as a test organism to identify potential interactions between full-length membrane proteins or between a full-length membrane protein and a soluble protein. The mbSUS can also be used to provide further evidence for protein-protein interactions detected with other methods and to map the interaction domains of selected proteins. The mbSUS is optimized for systematic screening approaches employing a mating-based approach, as typically used to determine protein interactions on a genomic scale. Construct... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926970 Mon, 01 Oct 2007 04:00:00 +0100 1926970 http://www.medworm.com/index.php?rid=1926963&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428666%26dopt%3DAbstract Authors: Bannon AW, Malmberg AB Experimental models of pain include tests of response thesholds to high intensity stimuli (acute pain tests) and changes in spontaneous or evoked behavioral responses in animals with peripheral injury or inflammation (persistent pain models). Acute thermal pain is modeled by the hot-plate and tail-flick test, while persistent pain can be modeled by the formalin test. This unit presents protocols for all three of these tests, including preparation of animals (rats or mice), administration of a compound being tested for its analgesic properties and data collection. PMID: 18428666 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926963 Mon, 01 Oct 2007 04:00:00 +0100 1926963 http://www.medworm.com/index.php?rid=1926961&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428668%26dopt%3DAbstract Authors: Cenci MA, Lundblad M This unit provides detailed protocols for establishing rodent models of L-DOPA-induced dyskinesia. The 6-hydroxydopamine (6-OHDA) lesion procedure is described in more detail for mice than for rats since the lesioning procedure in rats has been described extensively in previous work and is less difficult to perform. Unlike primate models, rodent models of L-DOPA-induced dyskinesia are relatively simple and fast to set up, thus being affordable to most laboratories. These models allow for studying the dyskinetic complications of L-DOPA treatment on large groups of animals under strictly controlled experimental conditions. Along with information and structured protocols for the practical execution of the test, this unit provides a detailed description of the... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926961 Mon, 01 Oct 2007 04:00:00 +0100 1926961 http://www.medworm.com/index.php?rid=1926974&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428655%26dopt%3DAbstract Authors: Fraefel C Herpes simplex virus type 1 (HSV-1)-based amplicon vectors contain only a very small percentage of the 152-kbp viral genome. Consequently, replication and packaging of amplicons depend on helper functions that are provided either by replication-defective mutants of HSV-1 or by replication-competent, but packaging-defective, HSV-1 genomes. Sets of cosmids that overlap and represent the entire HSV-1 genome can form, via homologous recombination, circular replication-competent viral genomes, which give rise to infectious virus progeny. However, if the DNA cleavage/packaging signals are deleted, reconstituted virus genomes are not packageable, but still provide all the helper functions required for the packaging of cotransfected amplicon DNA. The resulting stocks of pack... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926974 Sun, 01 Jul 2007 04:00:00 +0100 1926974 http://www.medworm.com/index.php?rid=1926972&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428657%26dopt%3DAbstract Authors: Mortensen R The analysis of mutant organisms and cell lines is important in determining the function of specific proteins. Recent technological advances in gene targeting by homologous recombination in mammalian systems enable the production of mutants in any desired gene, and can be used to produce mutant mouse strains and mutant cell lines. The yeast Flp/FRT recombinase system and bacteriophage recombinases such as Cre and its recognition sequence, loxP, allow spatial and temporal control of knockouts. This unit discusses crucial issues for homologous recombination experiments, including requirements for the source of DNA, criteria for the targeting constructs, methods of enrichment for homologous recombinants, (positive and negative selection, and the use of endogenous prom... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926972 Sun, 01 Jul 2007 04:00:00 +0100 1926972 http://www.medworm.com/index.php?rid=1926969&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428660%26dopt%3DAbstract Authors: Dodson SE, Heilman CJ, Kahn RA, Levey AI This unit details the use of bacterially produced fusion proteins for the production of antisera, allowing for the large-scale generation of affinity-purified antibodies to specific, targeted epitopes. The use of pET vectors containing a polyhistidine (His) or glutathione-S-transferase (GST) tag to construct bacterial expression plasmids are provided as prototypical examples of fusion protein methodology. The basic protocols provided in this unit describe: (1) transformation of E. coli for high-yield production of soluble fusion protein, (2) purification of soluble fusion proteins for use in immunization using chelated nickel or glutathione affinity chromatography (for His- and GST-tagged fusion proteins, respectively), (3) immunization... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926969 Sun, 01 Jul 2007 04:00:00 +0100 1926969 http://www.medworm.com/index.php?rid=1926965&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428664%26dopt%3DAbstract Authors: Pierce RC, Kalivas PW The study of locomotor activation in rodents does not involve extensive learning or conditioning, and so this dependent measure is often used as the initial screen for pharmacological effects predictive of therapeutic efficacy of a drug class in humans. Methods for evaluating locomotor behavior in rodents fall into two general categories: automated monitoring of behavior and direct observational techniques. This unit presents protocols for measurement of locomotor activity using photocell-based automated monitoring systems, and direct observation using interval and ordinal scales. Other methods used to quantify locomotor activity include video-based systems, rotometers, and running wheels; these are discussed in the commentary. The protocols provided are ... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926965 Sun, 01 Jul 2007 04:00:00 +0100 1926965 http://www.medworm.com/index.php?rid=1926975&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428654%26dopt%3DAbstract Authors: Oo TF, Burke RE Neuroscientists often need to detect neuron death at the light microscope level in tissue sections derived from animal models of neurological disease. In many instances there is a need to detect apoptosis, the most common morphology of programmed cell death. This unit provides two protocols for the detection of apoptosis by immunostaining for either activated forms of caspases or their cleavage products. When used in conjunction with nuclear dyes, these protocols permit visualization not only of caspase activation, but also the nuclear chromatin clumps characteristic of apoptosis. The first protocol utilizes peroxidase-mediated chromogen deposition to visualize antibodies by brightfield microscopy. The second protocol utilizes fluorophores to visualize antibodi... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926975 Sun, 01 Apr 2007 04:00:00 +0100 1926975 http://www.medworm.com/index.php?rid=1926971&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428658%26dopt%3DAbstract Authors: Crittenden JR, Heidersbach A, McManus MT RNA interference (RNAi) refers to the process by which 21- to 23-nucleotide short interfering RNAs (siRNAs) mediate post-transcriptional degradation of homologous mRNA transcripts. This process is carried out by an endogenous pathway that centers on the use of endogenously encoded small RNAs, and can be hijacked to knock down the expression of any target protein by introducing a specific siRNA into a cell. Stable knockdown can be obtained by constitutive expression of the siRNA from the host chromosome. Retroviruses, such as lentivirus, provide a convenient vector by which to integrate RNAi expression constructs. Lentiviruses can infect nondividing cells, thereby allowing knockdown in cells such as mature neurons. This unit provides met... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926971 Sun, 01 Apr 2007 04:00:00 +0100 1926971 http://www.medworm.com/index.php?rid=1926964&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428665%26dopt%3DAbstract Authors: Mar AC, Robbins TW Delay discounting refers to the degree to which immediate outcomes exhibit more influence over behavior than outcomes which are delayed. Impulsive choice, in the context of delay discounting, is generally considered as an increased preference for immediate over delayed outcomes, even where the delayed outcomes are more advantageous. In the past decade, there has been increasing use of delay-discounting paradigms to elucidate the physiological, pharmacological, and behavioral aspects of the putative neural circuitry underlying impulsive choice. This unit describes the assessment of impulsive choice in the rat using a delay-discounting procedure involving an operant response choice between a small reinforcer delivered immediately and a larger reinforcer delive... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926964 Sun, 01 Apr 2007 04:00:00 +0100 1926964 http://www.medworm.com/index.php?rid=1926962&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428667%26dopt%3DAbstract Authors: Powell SB, Geyer MA Animal models of schizophrenia may increase the understanding of the neurological abnormalities associated with the disorder and aid in the development of rational pharmacological treatments. Rather than attempting to model the entire syndrome of schizophrenia, a more biologically oriented approach to animal models has been to focus on specific symptoms of schizophrenia that are more objectively measured in the clinical population and more directly translatable to animals (e.g., observables or endophenotypes). This overview focuses on behavioral measures that have been investigated in rodent models of schizophrenia with varying degrees of predictive, etiological, and construct validity. Because of the severity of cognitive deficits in schizophrenia and thei... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926962 Sun, 01 Apr 2007 04:00:00 +0100 1926962 http://www.medworm.com/index.php?rid=1926976&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428653%26dopt%3DAbstract Authors: Phelan MC This unit opens with detailed discussions on the latest principles of sterile technique and preparation of culture media. Step-by-step protocols describe trypsinizing and subculturing monolayer cultures, passaging suspension cultures, freezing and thawing cells, counting cells using a hemacytometer, and preparing cells for transport. PMID: 18428653 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926976 Mon, 01 Jan 2007 05:00:00 +0100 1926976 http://www.medworm.com/index.php?rid=1926968&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428661%26dopt%3DAbstract Authors: DeWeese MR In vivo whole-cell patch-clamp recording provides a means for measuring membrane currents and potentials from individual cells in the intact animal. Patch-clamp methods have largely been developed in vitro. This body of work has contributed enormously to the understanding of many important phenomena in excitable cells--including synaptic plasticity in the mammalian central nervous system, and the behavior of individual protein channels. In recent years, an increasing number of groups have applied whole-cell recording techniques in the intact animal. Such in vivo studies offer the tantalizing possibility of uncovering the underlying principles and mechanisms of neural interactions within the natural context of fully intact biological networks. This unit focuses on st... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926968 Mon, 01 Jan 2007 05:00:00 +0100 1926968 http://www.medworm.com/index.php?rid=1926967&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428662%26dopt%3DAbstract Authors: Daw M, Isaac J Many brain processes and the expression of many neuronal proteins are developmentally regulated; therefore, studying cellular and molecular processes in tissue from very young animals is vital to understanding the development of the brain. Working on tissue from very young animals (first postnatal week) presents a number of specific problems. This unit describes techniques to overcome these problems to produce in vitro slices from neonatal rodents and to reliably achieve high-quality patch-clamp recordings from neurons in neonatal brain slices. PMID: 18428662 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926967 Mon, 01 Jan 2007 05:00:00 +0100 1926967 http://www.medworm.com/index.php?rid=1926966&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428663%26dopt%3DAbstract Authors: Deng C, Wang D, Bugaj-Gaweda B, De Vivo M Since the identification of phosphodiesterase activity in brain tissue more than 40 years ago, 11 distinct gene families have been identified, differing with respect to localization, regulation, affinity for cAMP and cGMP, and distinct functions within cells. PDEs 1, 2, 4, and 10 are currently of special interest to CNS pharmacology because of their high expression in specific areas of the brain and the behavioral effects of inhibitors of these enzymes in rodents. Efficient high-throughput PDE enzyme assays are essential for PDE-targeted drug discovery, and this unit details two types of assays. The first method is relatively inexpensive and is based on separating radiolabeled cNMPs from degradation products on alumina columns. The sec... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926966 Mon, 01 Jan 2007 05:00:00 +0100 1926966 http://www.medworm.com/index.php?rid=1926997&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428632%26dopt%3DAbstract Authors: Raju DV, Smith Y The mammalian brain contains a myriad of interconnected regions. An examination of the complex circuitry of these areas requires sensitive neuroanatomical tract tracing techniques. The anterograde tracers, Phaseolus vulgaris leucoagglutinin (PHA-L) and biotinylated dextran amines (BDA) are powerful tools that can be used to label fiber tracts that project from one particular brain region. When injected iontophoretically, PHA-L and BDA are readily taken up by neurons and transported anterogradely along their axonal tracts. Combined with immunocytochemistry for neurotransmitters, neuropeptides, and receptors, tract tracing methods may be used to elucidate the phenotype of synapses that form the microcircuitry of specific neural systems. PMID: 18428632 [PubMe... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926997 Wed, 01 Nov 2006 05:00:00 +0100 1926997 http://www.medworm.com/index.php?rid=1926994&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428635%26dopt%3DAbstract Authors: Cameron HA Injection and immunohistochemical detection of 5-bromo-2'-deoxyuridine (BrdU) has become the standard method for studying the birth and survival of neurons, glia, and other cell types in the nervous system. BrdU, a thymidine analog, becomes stably incorporated into DNA during the S-phase of mitosis. Because DNA containing BrdU can be specifically recognized by antibodies, this method allows dividing cells to be marked at any given time and then identified at time points from a few minutes to several years later. BrdU immunohistochemistry is suitable for cell counting to examine the regulation of cell proliferation and cell fate. It can be combined with labeling by other antibodies, allowing confocal analysis of cell phenotype or expression of other proteins. The pot... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926994 Wed, 01 Nov 2006 05:00:00 +0100 1926994 http://www.medworm.com/index.php?rid=1926992&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428637%26dopt%3DAbstract Authors: Salmon P, Trono D Lentiviral vectors have emerged over the last decade as powerful, reliable and safe tools for stable gene transfer in a wide variety of mammalian cells. Unlike other vectors derived from oncoretroviruses, they allow for stable gene delivery into most nondividing primary cells, including neurons. This is why LVs are becoming the most useful and promising tools in the field of neuroscience, not only for research, but also for future gene and cell therapy approaches. Lentivectors (LVs) derived from HIV-1 have gradually evolved to display many desirable features aimed at increasing both their safety and their versatility. These latest designs are reviewed in this unit. This unit also describes protocols for production and titration of LVs that can be implemented ... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926992 Wed, 01 Nov 2006 05:00:00 +0100 1926992 http://www.medworm.com/index.php?rid=1926987&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428642%26dopt%3DAbstract Authors: Kao JP A caged molecule is an inert but photosensitive molecule that is transformed by photolysis into a biologically active molecule at high speed (typically 1 msec). The process is referred to as photorelease. The spatial resolution of photorelease is limited by the properties of light; submicrometer resolution is potentially achievable. Therefore, focal photorelease of caged molecules enables one to control biological processes with high spatio-temporal precision. The principles underlying caged molecules as well as practical considerations for their use are discussed in this unit. PMID: 18428642 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926987 Wed, 01 Nov 2006 05:00:00 +0100 1926987 http://www.medworm.com/index.php?rid=1926986&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428643%26dopt%3DAbstract Authors: Tang CM Photolysis of "caged" compounds is a technique for releasing biologically active compounds in which the timing, rate, and spatial profile of release are controlled by light. Issues relating to the delivery of light for single-photon photolysis are presented. Specific discussions include the theories relating to how light interacts with biological tissue to produce scattering and phototoxicity, as well as the issues involved in choosing the appropriate light source. Several approaches and optical designs are presented for delivering the output of a laser to a microscopic specimen. The criteria for choosing an approach are presented. The commercial sources for the parts needed to build a photolysis system are also provided. This unit will be particularly useful for inves... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926986 Wed, 01 Nov 2006 05:00:00 +0100 1926986 http://www.medworm.com/index.php?rid=1926996&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428633%26dopt%3DAbstract Authors: Kim SH, Che P, Chung SH, Doorn D, Hoy M, Larouche M, Marzban H, Sarna J, Zahedi S, Hawkes R The gross anatomical distribution of an antigen is typically mapped using a combination of serial sectioning, immunocytochemistry, and three-dimensional reconstruction. This is a tedious and time-consuming procedure, which introduces an array of potential alignment and differential shrinkage errors and requires considerable experience and specialized equipment. In particular, it is unsuited for routine screening applications. To circumvent these problems, this unit presents a routine whole-mount immunocytochemistry protocol that can be used to map many antigenic distributions in the developing and adult brain. The technique can also be easily adapted to detect anterograde and retrograde... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926996 Tue, 01 Aug 2006 04:00:00 +0100 1926996 http://www.medworm.com/index.php?rid=1926995&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428634%26dopt%3DAbstract Authors: Hall AK Sensory neurons have proven very useful for analysis of neuronal differentiation in vivo and in vitro. Their utility for in vitro work is based on the fact that sensory neurons are relatively easy to isolate in large numbers and are amenable to manipulations in culture. Lumbar ganglia are usually used because their location in the caudal nervous system means they are the least differentiated at any developmental stage, allowing the analysis of relatively undifferentiated cells. Rodent sensory ganglia from embryonic to adult stages can be dissected effectively and maintained in serum-free medium or in coculture with other cells or factors. This unit describes generation of embryonic rat lumbar dorsal root ganglia (DRG) cultures, which form an important model system for ... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926995 Tue, 01 Aug 2006 04:00:00 +0100 1926995 http://www.medworm.com/index.php?rid=1926980&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428649%26dopt%3DAbstract Authors: Boggiano MM, Chandler PC This unit describes a rodent model of binge eating based on cyclic restriction, refeeding, footshock, and intermittent access to palatable food. These conditions mimic dieting, stress, and "junk" food indulgence, respectively, all common etiological and maintenance factors in human binge eating. Four groups of rats are used: one subjected to cyclic food restriction, another to acute footshock stress, another to both of these (R + S), and a control. Neither cyclic restriction nor stress alone produces binge eating, but the R + S rats, despite satiety, double their intake of palatable food in a discrete period of time (i.e., binge) when stressed. This protocol recapitulates critical properties of human binge eating, namely preference for palatable food, ... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926980 Tue, 01 Aug 2006 04:00:00 +0100 1926980 http://www.medworm.com/index.php?rid=1926979&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428650%26dopt%3DAbstract Authors: Corwin RL, Wojnicki FH In this protocol, binge-type eating is induced in non-food-deprived rats by providing limited access to an optional source of dietary fat: vegetable shortening. The protocol is simple and inexpensive, and the binge behavior is robust, reliable, and maintainable across extended periods of time. Two peptides that normally affect fat intake in rats have no effect on fat intake under limited-access conditions. However, recent results with a GABA(B) receptor agonist and with progressive-ratio responding suggest that the behavior induced by the limited-access binge protocol may share similarities with substance abuse. This protocol is designed to model the kind of excessive behavior that characterizes bingeing-related eating disorders and certain addictions. ... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926979 Tue, 01 Aug 2006 04:00:00 +0100 1926979 http://www.medworm.com/index.php?rid=1926978&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428651%26dopt%3DAbstract Authors: Avena NM, Rada P, Hoebel BG Bingeing behavior is characteristic of many eating disorders. This unit describes an animal model of sugar bingeing. This model has been used successfully to elicit behavioral and neurochemical signs of sugar dependence in rats, e.g., indices of bingeing, withdrawal, increased intake after abstinence (deprivation effect), cross-sensitization with amphetamine, and increases in dopamine release in the nucleus accumbens due to repeated bingeing. PMID: 18428651 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926978 Tue, 01 Aug 2006 04:00:00 +0100 1926978 http://www.medworm.com/index.php?rid=1926977&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428652%26dopt%3DAbstract Authors: Alberts P, Johansson BG, McArthur RA The need for treatment of obesity and obesity-related diseases, such as type 2 diabetes, has been intensified by the epidemic rise of obesity. Recent advances make possible continuous monitoring of metabolically relevant functions in animals to identify novel thermogenic and anorectic compounds. This unit describes non-invasive in vivo calorimetric assessment of energy expenditure using measurements of oxygen consumption and carbon dioxide production, complemented by telemetric monitoring of body core temperature and locomotor activity in mice and rats. Reference compounds are used to illustrate the determination of substance-specific parameters, such as the dose that produces the half-maximal effect (ED(50)), the maximal effect, as well as... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926977 Tue, 01 Aug 2006 04:00:00 +0100 1926977 http://www.medworm.com/index.php?rid=1926993&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428636%26dopt%3DAbstract Authors: Choi VW, Asokan A, Haberman RA, McCown TJ, Samulski RJ Adeno-associated virus is a nonpathogenic human virus that has been developed into a gene-delivery vector due to its high efficiency of infection for many different cell types and its ability to persist and lead to long-term gene expression. This unit describes efficient methods to generate high-titer, research-grade, adenovirus-free recombinant single-stranded and self-complementary adeno-associated virus in various serotypes, along with methods to quantify the viral vectors. Two detailed methods are provided for viral vector injection into rat brain and for histological detection of transgene expression, including lacZ and GFP. PMID: 18428636 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926993 Mon, 01 May 2006 04:00:00 +0100 1926993 http://www.medworm.com/index.php?rid=1926989&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428640%26dopt%3DAbstract Authors: Bonifacino JS, Dell'Angelica EC, Springer TA Immunoprecipitation is a technique in which an antigen is isolated by binding to a specific antibody attached to a sedimentable matrix. It is also used to analyze protein fractions separated by other biochemical techniques such as gel filtration or density gradient sedimentation. The source of antigen for immunoprecipitation can be unlabeled cells or tissues, metabolically or intrinsically labeled cells, or in vitro-translated proteins. This unit describes a wide range of immunoprecipitation techniques, using either suspension or adherent cells lysed by various means (e.g., with and without detergent, using glass beads, etc.). Flow charts and figures give the user a clear-cut explanation of the options for employing the technology. ... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926989 Mon, 01 May 2006 04:00:00 +0100 1926989 http://www.medworm.com/index.php?rid=1926988&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428641%26dopt%3DAbstract Authors: Elion EA Coprecipitation of proteins from whole-cell extracts is a valuable approach to testing for physical interactions between proteins of interest. When a precipitating antibody is used, this method is referred to as co-immunoprecipitation. Coprecipitation can be used to study interactions between known proteins and as a means of identifying components of a complex. This unit describes basic approaches to immunoprecipitating tagged proteins from whole-cell extracts. The extract is prepared under nondenaturing conditions, the protein of interest is precipitated, and the precipitate is tested for a second specifically associated protein. The approach can be used for native or epitope-tagged proteins for which antibodies are available, or for recombinant proteins that bind wi... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926988 Mon, 01 May 2006 04:00:00 +0100 1926988 http://www.medworm.com/index.php?rid=1926991&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428638%26dopt%3DAbstract Authors: Wouters FS, Bastiaens PI Detection of specific protein-protein interactions has long been restricted to bulk biochemical methods such as immunoprecipitation and immunoblotting. Even more sensitive methods using general immunofluorescence are limited, and it is difficult to infer protein-protein interactions from the results of these tests. Fluorescence Resonance Energy Transfer (FRET) is a photophysical process that can be exploited to obtain highly sensitive information about such interactions. It can sense the presence of acceptor fluorophores in the vicinity of a donor fluorophore within a separation distance that is the size of a single protein molecule. This unit details FRET microscopy based on release of quenched donor fluorescence after acceptor photobleaching, microin... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926991 Wed, 01 Feb 2006 05:00:00 +0100 1926991 http://www.medworm.com/index.php?rid=1926990&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428639%26dopt%3DAbstract Authors: Hamdan FF, Percherancier Y, Breton B, Bouvier M Bioluminescence resonance energy transfer (BRET) allows monitoring of protein-protein interactions in real time in living cells. One candidate interacting protein is fused to a luminescent energy donor, such as Renilla luciferase, and the other to a fluorescent energy acceptor, such the green fluorescent protein (GFP), and the two are then coexpressed in the same cells. If the two proteins interact, their close proximity allows nonradiative energy transfer (BRET) between the luciferase and the GFP. BRET does not occur if the two proteins are separated by more than 100 A, making the technique ideal for monitoring protein-protein interactions in biological systems. This unit describes the use of BRET to study constitutive and agoni... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926990 Wed, 01 Feb 2006 05:00:00 +0100 1926990 http://www.medworm.com/index.php?rid=1926985&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428644%26dopt%3DAbstract Authors: Willott JF The three general approaches for assessing the mouse auditory system presented in UNITS 8.21B-8.21D are complementary, but differ in the time required for testing as well as the goals of the tests: the auditory brainstem response (ABR; UNIT 8.21B), distortion product otoacoustic emissions (DPOAE; UNIT 8.21C), and lick suppression behavioral test (UNIT 8.21D). Various issues that need to be addressed in the testing of mouse hearing are discussed in this overview, including accurate nomenclature of the strain and/or mutations, environmental versus genetic influences, selection of frequencies to be tested, and appropriate animal care and use. PMID: 18428644 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926985 Wed, 01 Feb 2006 05:00:00 +0100 1926985 http://www.medworm.com/index.php?rid=1926984&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428645%26dopt%3DAbstract Authors: Willott JF The ABR is an electroencephalographic response measured with scalp electrodes. It provides a quick, easy, and reliable method for physiological assessment of auditory sensitivity in mice. A series of brief tone pips or clicks is presented to an anesthetized mouse at a high rate of speed; each click evokes waves of neural activity in the brainstem that are computer-averaged so they are differentiated from non-auditory background voltages. The intensity of the clicks is reduced in steps until an ABR can no longer be discerned, thereby defining the ABR threshold, which is closely related to the hearing threshold. Key procedural issues are: (1) accurate calibration of the acoustics (what sounds arrive at the mouse's ear), (2) anesthetization of the mouse, (3) setting up... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926984 Wed, 01 Feb 2006 05:00:00 +0100 1926984 http://www.medworm.com/index.php?rid=1926983&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428646%26dopt%3DAbstract Authors: Martin GK, Stagner BB, Lonsbury-Martin BL Distortion-product otoacoustic emissions (DPOAEs) can be measured in the ear canal following the presentation of two tones. These emissions are generated by the outer hair cells (OHCs) of the inner ear and they are reduced or absent when the OHCs are damaged by, for example, exposure to excessive noise or ototoxic drugs. Consequently, DPOAEs provide a powerful and noninvasive means to assess the robustness of OHC function. A detailed method is described for measuring DPOAEs to assess cochlear function in mice. Recommendations are given for the required equipment and instructions are presented for setting up a DPOAE system. Also, a protocol is outlined for measuring DPOAEs in mice and troubleshooting tips are provided. Examples of data ... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926983 Wed, 01 Feb 2006 05:00:00 +0100 1926983 http://www.medworm.com/index.php?rid=1926982&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428647%26dopt%3DAbstract Authors: Heffner HE, Koay G, Heffner RS The method of conditioned suppression described in this unit involves training a thirsty mouse to make steady contact with a waterspout in order to receive a slow, but steady trickle of water and then pairing a sound with mild electric shock delivered through the spout. The mouse quickly learns to avoid the shock by breaking contact with the spout whenever it detects the sound. This suppression of drinking is then used to indicate that the animal detected the sound. Because it is a cognitively simple procedure, conditioned suppression can be used to determine the ability of brain-damaged and genetically altered as well as normal animals to detect and discriminate sounds. PMID: 18428647 [PubMed - indexed for MEDLINE] (Source: Current Protocols... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926982 Wed, 01 Feb 2006 05:00:00 +0100 1926982 http://www.medworm.com/index.php?rid=1926981&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428648%26dopt%3DAbstract Authors: Heinrichs SC, Koob GF This unit presents eight separate stressor protocols for laboratory rodents. Stress induction is a critical element in the study of neural and neuroendocrine mechanisms involved in establishing and maintaining a state of stress. The first four procedures, immobilization, footshock, swimming, and noise, involve acute exposure to noxious stimuli. The next three procedures, social isolation, resident/intruder aggression, and maternal deprivation, induce social disruption by withdrawal from a group housing condition, attack within the unfamiliar territory of a dominant male, or segregation of a preweanling pup from its mother, respectively. The final procedure, sleep deprivation, involves passive denial of the opportunity to sleep. Support protocols are provi... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926981 Wed, 01 Feb 2006 05:00:00 +0100 1926981 http://www.medworm.com/index.php?rid=1927013&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428616%26dopt%3DAbstract Authors: Donovan J, Brown P This unit describes the techniques for the following routes of injection for mice, rats, hamsters and rabbits: intramuscular, intradermal, subcutaneous, intravenous, intraperitoneal, footpad, and intrathymic. Guidelines are also given regarding injection volumes and temperatures, and the use of proper restraints. PMID: 18428616 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927013 Tue, 01 Nov 2005 05:00:00 +0100 1927013 http://www.medworm.com/index.php?rid=1927012&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428617%26dopt%3DAbstract Authors: Donovan J, Brown P Blood is most frequently sampled for evaluation of serum antibodies or analysis of surface markers on peripheral blood cells. The protocols offered in this unit describe collection of blood from the orbital sinus or plexus of the mouse, rat, or hamster. With appropriate techniques, small amounts of blood can be obtained with little ill effect on the animal. Collection from the auricular vein or artery of the rabbit is also relatively unstressful to the animal. Bleeding procedures that should be performed on the anesthetized animal include collection from the mouse axillary plexus, cardiac puncture of the mouse, rat, hamster, or rabbit, and collection from the hamster abdominal aorta or vena cava. PMID: 18428617 [PubMed - indexed for MEDLINE] (Source: Cur... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927012 Tue, 01 Nov 2005 05:00:00 +0100 1927012 http://www.medworm.com/index.php?rid=1927011&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428618%26dopt%3DAbstract Authors: Donovan J, Brown P Euthanasia of laboratory animals must be performed by trained personnel using appropriate techniques, equipment, and reagents in order to effect a death that is humane and satisfies research requirements. Acceptable methods of euthanasia are painless or minimize distress, and are quick and easy to perform, safe for those performing the procedure, and efficient and economic. They are aesthetically acceptable and are done in the absence of other animals. In addition, these methods do not result in gross histological or histochemical changes that would adversely affect research results. This unit offers protocols for euthanasia employing carbon dioxide asphyxiation (see Basic Protocol 1), pentobarbital overdose (see Basic Protocol 2), exsanguination, and cervic... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927011 Tue, 01 Nov 2005 05:00:00 +0100 1927011 http://www.medworm.com/index.php?rid=1927008&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428621%26dopt%3DAbstract Authors: Maric D, Barker JL Neural stem cells (NSCs) are defined as undifferentiated cells originating from the neuroectoderm that have the capacity both to perpetually self-renew without differentiating and to generate multiple types of lineage-restricted progenitors (LRPs). LRPs can themselves undergo limited self-renewal and ultimately differentiate into highly specialized cells that make up the nervous system. However, this physiologically delimited definition of NSCs and LRPs has become increasingly blurred due to lack of protocols for effectively separating these types of cells from primary tissues. This unit discusses recent attempts using fluorescence-activated cell sorting (FACS) strategies to prospectively isolate NSCs from different types of LRPs as they appear in vivo, and ... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927008 Tue, 01 Nov 2005 05:00:00 +0100 1927008 http://www.medworm.com/index.php?rid=1927003&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428626%26dopt%3DAbstract Authors: Slotnick B, Restrepo D A computer-controlled, multiple-channel odor generator, operant test chamber, and discrimination training procedure for mice are described. The odor generator allows controlled presentation of any one or combinations of eight odors to an odor-sampling port that contains a liquid reinforcement delivery tube that also serves to detect responses. A modified discrete trial operant conditioning procedure provides measures of both response accuracy and response rate during anticipation of stimulus delivery and in the presence of two different odor stimuli. Results from numerous experiments demonstrate that, with these methods, mice reliably and rapidly acquire odor detection and a variety of odor-discrimination tasks, and that response rate in the presence of ... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927003 Tue, 01 Nov 2005 05:00:00 +0100 1927003 http://www.medworm.com/index.php?rid=1926998&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428631%26dopt%3DAbstract Authors: Dodart JC, May P In Alzheimer's disease (AD), characteristic lesions develop in brain regions that subserve cognitive functions, ultimately leading to dementia. There are now several lesioned or transgenic small-animal models of the disease that model select aspects of cognitive deficits and/or recapitulate many, but not all, of the characteristic pathologic lesions observed in AD. This overview describes the most common approaches used to model AD in rodents, highlights their utility, and discusses some of their deficiencies. PMID: 18428631 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926998 Tue, 01 Nov 2005 05:00:00 +0100 1926998 http://www.medworm.com/index.php?rid=1927010&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428619%26dopt%3DAbstract Authors: Wouterlood FG A confocal laser scanning microscope (CLSM) collects information from a thin, focal plane and ignores out-of-focus information. The operator configures separate channels (laser, filters, detector settings) for each fluorochrome used in a particular experiment. Then, 3-D reconstructions are made from Z-series of confocal images: one series per channel. Channel signal separation is extremely important and measures to avoid bleaching are vital. Post-acquisition deconvolution of the image series is then performed to increase resolution. In the 3-D reconstruction program described in this unit, reconstructions can be inspected in real time from any viewing angle. By altering viewing angles and by switching channels off and on, the spatial relationship of 3-D-reconstru... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927010 Mon, 01 Aug 2005 04:00:00 +0100 1927010 http://www.medworm.com/index.php?rid=1927009&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428620%26dopt%3DAbstract Authors: Belluscio L Two-photon imaging is an innovative optical technique that has quickly become state of the art for imaging fluorescent signals in a variety of organisms. With many advantages over conventional confocal microscopy, such as greater image resolution, deeper access (approximately 400 microm), and much less photo damage, two-photon microscopy has already proven to be an extremely useful tool for imaging live cells or tissue. Due to its tremendous versatility, recent efforts have adapted this technique to allow visualization of fluorescent cells directly in living animals. This unit describes a basic procedure for performing two-photon imaging in vivo as applied to the dorsal surface of the brain in live anesthetized mice or rats. The protocol outlines a surgical prepara... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927009 Mon, 01 Aug 2005 04:00:00 +0100 1927009 http://www.medworm.com/index.php?rid=1927000&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428629%26dopt%3DAbstract Authors: Thomsen M, Caine SB Chronic intravenous drug self-administration in rodents is a useful procedure for predicting the abuse liability of novel drugs in humans, for evaluating candidate treatments for drug abuse and dependence, and for studying the biological basis of addiction. This unit focuses on recent technical innovations for conducting long-term studies of i.v. drug self-administration behavior in healthy, freely moving rats and mice. Included are protocols for construction, implantation, and maintenance of chronic indwelling jugular catheters, commentary on critical parameters, a troubleshooting guide, and anticipated results. PMID: 18428629 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927000 Mon, 01 Aug 2005 04:00:00 +0100 1927000 http://www.medworm.com/index.php?rid=1926999&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428630%26dopt%3DAbstract Authors: Platt DM, Carey G, Spealman RD Drug self-administration is a procedure in which a subject performs a response, called an operant, that results in the delivery of a drug injection. This procedure is viewed as a relevant model for the study of human drug-taking behavior. Drug self-administration in primates has several characteristics that resemble drug-taking behavior in humans, and drugs that are commonly abused by humans also typically maintain self-administration behavior in monkeys. Drug self-administration procedures allow for the study of a variety of drug properties. For instance, they are used to investigate the abuse potential of new compounds and to study the effects of candidate medications for the treatment of drug addiction. These procedures also can be used to stu... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1926999 Mon, 01 Aug 2005 04:00:00 +0100 1926999 http://www.medworm.com/index.php?rid=1927007&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428622%26dopt%3DAbstract Authors: Yang XW, Gong S This unit provides a comprehensive overview on the generation of transgenic mice using bacterial artificial chromosomes (BACs), and the application of BAC transgenic mice in neuroscience research. In the first section, advantages of the BAC transgenic approach compared to the conventional transgenic approach are summarized. In the second section, important considerations in designing BAC transgenic constructs are outlined. Four commonly used BAC transgenic construct designs are also outlined. Concepts of modifying BACs by homologous recombination in E. coli to introduce a variety of mutations into BACs, and important steps to characterize a modified BAC prior to the generation of transgenic mice are also presented. In the final section, some of the important ap... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927007 Sun, 01 May 2005 04:00:00 +0100 1927007 http://www.medworm.com/index.php?rid=1927006&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428623%26dopt%3DAbstract Authors: Gong S, Yang XW BAC transgenesis is a powerful tool for the study of gene expression and gene function in the mouse in vivo. In this unit, detailed protocols are provided for modification (i.e., marker gene insertion, deletion, or point mutation) of BACs by homologous recombination in E. coli. This method utilizes a shuttle vector that allows transient expression of the E. coli RecA gene to support homologous recombination in the BAC host bacteria. In addition, two protocols are provided for purification of BAC DNA for microinjection to generate transgenic mice. Since BAC DNA is prone to degradation, which may introduce positional effects in transgenic mice, two methods are given for purification of intact BAC DNA for subsequent microinjection. PMID: 18428623 [PubMed - ind... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927006 Sun, 01 May 2005 04:00:00 +0100 1927006 http://www.medworm.com/index.php?rid=1927002&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428627%26dopt%3DAbstract Authors: Humby T, Wilkinson L, Dawson G First developed in the 1980s by Trevor Robbins in Cambridge to investigate attentional function in rats, the 5-choice serial reaction time task involves continuous scanning by the subject across a spatial array of visual stimuli. On detecting a brief stimulus at one of five locations, the subject must make a nose poke in order to collect a reward. The task has also been conducted successfully in several strains of mice, including transgenic models. This unit presents the procedures required to run the task in mice, outlining the expected results at each stage of training and presenting examples of the most common manipulations used to dissociate behavior further. PMID: 18428627 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neur... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927002 Sun, 01 May 2005 04:00:00 +0100 1927002 http://www.medworm.com/index.php?rid=1927001&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428628%26dopt%3DAbstract Authors: Hellier JL, Dudek FE Animal models of injury-induced epilepsy may provide insight into the mechanisms of acquired epilepsy. Previous animal models of temporal lobe epilepsy (TLE) were produced by acute treatments that often have high mortality rates and/or are associated with a low proportion of animals developing spontaneous, chronic motor seizures. In this unit, a protocol is provided for inducing chronic epilepsy in rats using multiple, low-dose, intraperitoneal injections of an excitotoxic agent, kainic acid. This protocol reliably induces TLE in nearly all treated rats (97% had at least two observed spontaneous motor seizures) with a relatively low mortality rate (<15%). This modified chemoconvulsant treatment protocol (i.e., multiple low doses) is efficient and relati... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927001 Sun, 01 May 2005 04:00:00 +0100 1927001 http://www.medworm.com/index.php?rid=1927005&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428624%26dopt%3DAbstract Authors: Kahl SD, Felder CC Scintillation proximity assay technologies provide a rapid non-separation method to measure common biological interactions using radioactively tagged molecules. This unit identifies potential uses of the technology for the measurement of receptor-ligand binding, cAMP accumulation, GTP binding to heterotrimeric G proteins, protease activity and cellular uptake. PMID: 18428624 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927005 Tue, 01 Feb 2005 05:00:00 +0100 1927005 http://www.medworm.com/index.php?rid=1927004&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428625%26dopt%3DAbstract Authors: Blanchard DC, Blanchard RJ, Griebel G Defensive responses include an array of specific behaviors, including flight, freezing, risk assessment, and defensive threat/attack, that are elicited by unconditioned threat stimuli such as predators or predator odors. Some individual defensive behaviors are selectively responsive to drugs effective against generalized anxiety disorder or panic, providing a rationale for their use in investigation of compounds that may be useful in treating these disorders. In addition, defensive behaviors toward predators and some predator odors show rapid conditioning to contextual stimuli, whereas other predator odors do not, although they too elicit defensiveness. This pattern suggests that the ability of a predator odor to predict danger may be a de... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927004 Tue, 01 Feb 2005 05:00:00 +0100 1927004 http://www.medworm.com/index.php?rid=1927028&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428601%26dopt%3DAbstract Authors: Gallagher S, Winston SE, Fuller SA, Hurrell JG Immunoblotting (western blotting) is used to identify specific antigens recognized by polyclonal or monoclonal antibodies. This unit provides numerous protocols for all steps starting with solubilization of the protein samples, usually with SDS and reducing agents. Following solubilization, the material is separated by SDS-PAGE and the antigens are then electrophoretically transferred to a membrane, a process that can be monitored by reversible staining or Ponceau S staining. The transferred proteins are bound to the surface of the membrane, providing access to immunodetection reagents. Any remaining binding sites are blocked by immersing the membrane in a blocking solution. After probing with the primary antibody, the membrane is... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927028 Mon, 01 Nov 2004 05:00:00 +0100 1927028 http://www.medworm.com/index.php?rid=1927026&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428603%26dopt%3DAbstract Authors: Poolos NP, Jones TD Pyramidal neurons of the central nervous system have extensively arborized apical dendrites that contribute importantly to the signaling properties of the neuron. Recent advances in electrophysiological techniques have allowed recording from neuronal dendrites. These techniques depend on using infrared optics to visualize dendritic processes in the unstained brain slice preparation, on pipet positioning with high resolution micromanipulators, and on stringent techniques for brain slice preparation that preserved healthy dendritic processes, even in tissue from mature animals. The procedures underlying these techniques are described in this unit. PMID: 18428603 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927026 Mon, 01 Nov 2004 05:00:00 +0100 1927026 http://www.medworm.com/index.php?rid=1927018&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428611%26dopt%3DAbstract Authors: Goodson S, Halford J, Blundell J Analysis of rodent feeding behavior during the study of anorectic drugs reveals that contextual variables such as type of food, drug, environment, and physiological state influence food intake. Temporal changes in behavior can signal the onset of a feeding modulation process, as is the case with anorectic drugs acting on distinct neurochemical mechanisms that change temporal behavioral profiles. Comparing the effects of anorectic agents in controlled animal assays can identify potential therapeutic drugs that may influence human eating behavior. This unit specifies the materials and procedures required for continuously monitoring the Behavioral Satiety Sequence, as well as a device for continuous, automatic measurement of the mass of food consu... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927018 Mon, 01 Nov 2004 05:00:00 +0100 1927018 http://www.medworm.com/index.php?rid=1927017&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428612%26dopt%3DAbstract Authors: Riley AL, Freeman KB Administration of a drug following ingestion of a novel food or solution often suppresses subsequent intake of the new food or solution. This suppression is associative, in that consumption is not suppressed when there is no temporal relationship between consumption and drug administration. The robust nature of aversion learning has made this procedure a sensitive and widely used behavioral index of drug side effects. The procedures described in this unit are suitable for work with rodents, and may require modifications, e.g., in presentation of the ingesta and drug for other species. Familiar and novel foods may be used instead of solutions, with similar results. PMID: 18428612 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927017 Mon, 01 Nov 2004 05:00:00 +0100 1927017 http://www.medworm.com/index.php?rid=1927015&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428614%26dopt%3DAbstract Authors: Morrow TJ Painful peripheral neuropathy is a common secondary complication of diabetes. The streptozotocin (STZ)-induced diabetic rat is the most commonly employed animal model used to study mechanisms of painful diabetic neuropathy and to evaluate potential therapies. A low dose STZ protocol is described for inducing experimental diabetes in the rat. Several behavioral assays are described, which are routinely used to assess different aspects of neuropathic pain in this animal model of diabetes mellitus, including mechanical allodynia and heat hyperalgesia. PMID: 18428614 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927015 Mon, 01 Nov 2004 05:00:00 +0100 1927015 http://www.medworm.com/index.php?rid=1927038&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428591%26dopt%3DAbstract Authors: Donovan J, Brown P For the safety of the handler and the animal, proper methods for handling and restraining laboratory animals should be followed. Improper handling can result in increased stress and injury to the animal. In addition, the handler risks injury from bite wounds or scratches inflicted when the animal becomes fearful or anxious. By using sure, direct movements with a determined attitude, the animal can be easily handled and restrained. Animals can be restrained either manually or in a plastic restrainer. The protocols in this unit describe handling and manual restraint of mice, rats, hamsters, and rabbits. Alternate protocols describe restraint using the plastic restrainer. PMID: 18428591 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscien... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927038 Wed, 01 Sep 2004 04:00:00 +0100 1927038 http://www.medworm.com/index.php?rid=1927037&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428592%26dopt%3DAbstract Authors: Donovan J, Brown P Proper identification of animals is crucial in research, whether the animals are being used as experimental subjects, for breeding purposes, or as a source for tissues, cells, or fluids. This unit provides specific protocols for the ear punch (notch) and ear tag methods for mouse, rat and hamster; tattoos for mouse, rat and rabbit; and the correct use of a subcutaneous transponder system for all of the above species. PMID: 18428592 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927037 Wed, 01 Sep 2004 04:00:00 +0100 1927037 http://www.medworm.com/index.php?rid=1927036&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428593%26dopt%3DAbstract Authors: Brody T, Odenwald W The goal of neurogenetics is an understanding of the genetic basis of brain structure and function. Neurogenetics deals with multiple areas of investigation, including the genetic basis of neural induction, patterning, cell fate specification, neuron maturation, axonal and dendritic organization, synapse function, and learning and behavior. This appendix provides links to databases and other Web sites used by neurobiologists for recovery of information about genes involved in neural development and neuron function. Special care has been taken to curate sites involving model organisms, since neurogenetics relies heavily on comparative genomics to establish gene function. PMID: 18428593 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neurosci... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927036 Wed, 01 Sep 2004 04:00:00 +0100 1927036 http://www.medworm.com/index.php?rid=1927034&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428595%26dopt%3DAbstract Authors: Finkelstein DI, Stanic D, Parish CL, Drago J, Horne MK Stereological methods allow for the determination of cell numbers, terminal densities, and, subsequently, the estimation of terminal arbor size within a given brain nucleus. This unit provides an explanation for determining the terminal arbor size of dopaminergic neurons of the nigrostriatal pathway in rodents. In contrast to previously used single-axon reconstructions, these stereological methods allow for quick and easy determination of terminal arbor size. PMID: 18428595 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927034 Wed, 01 Sep 2004 04:00:00 +0100 1927034 http://www.medworm.com/index.php?rid=1927031&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428598%26dopt%3DAbstract Authors: Leenders M, Gerwin C, Sheng ZH Investigation of synaptic vesicle membrane proteins using multidisciplinary approaches, particularly to characterize synaptic vesicle proteins in synapses, can greatly advance our knowledge of the molecular mechanisms involved in synaptic vesicle exocytosis and neurotransmission. Three approaches are presented in this unit to identify and characterize synaptic vesicle proteins. The first is a subcellular fractionation method used to isolate synaptic vesicles from rat brain synaptosomal preparations, which can then be used in a variety of biochemical studies on synaptic vesicle proteins. The second is a detailed procedure for pre-embedding immunogold staining and electron microscopic observation, techniques that permit the morphological identifica... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927031 Wed, 01 Sep 2004 04:00:00 +0100 1927031 http://www.medworm.com/index.php?rid=1927029&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428600%26dopt%3DAbstract Authors: Schetz JA, Shankar EP The Schneider-2 (S2) Drosophila cell line is well suited for the stable overexpression of recombinant proteins using plasmid-based protein expression vectors. Following drug selection, a polyclonal S2 cell line can be induced to express on the order of 2 to 100 pmol/mg membrane protein for G-coupled protein receptors, 4000 to 100,000 sites/cell for other membrane receptors and 3 to 35 mg/liter for soluble and secreted proteins. PMID: 18428600 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927029 Wed, 01 Sep 2004 04:00:00 +0100 1927029 http://www.medworm.com/index.php?rid=1927021&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428608%26dopt%3DAbstract Authors: Wehner JM, Radcliffe RA Contextual and cued fear conditioning is a robust form of learning in which an association is made between stimuli and their aversive consequences. Fear conditioning has been used in laboratory rodents in part because it is a highly conserved form of behavior that is exhibited in both laboratory situations and in normal environments. Training requires only a single trial and this makes it adaptable to genetic, pharmacological, and biochemical studies. Clinically, it is has relevance to human behavior in that fear conditioning can be produced in humans, and damage to the amygdala prevents fear conditioning. PMID: 18428608 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927021 Wed, 01 Sep 2004 04:00:00 +0100 1927021 http://www.medworm.com/index.php?rid=1927019&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428610%26dopt%3DAbstract Authors: Wrenn CC The social transmission of food preference (STFP) is an ethologically relevant test of olfactory memory that can be used in mice. In this test, "observer" mice interact with a "demonstrator" mouse that has recently eaten a novel food. When observer mice are presented with a choice between the food eaten by the demonstrator and some other novel food, observer mice prefer the food eaten by the demonstrator. This phenomenon depends on the observer mice detecting olfactory cues on the breath of the demonstrator mouse during their interaction. The subsequent food preference serves as a measure of memory for those olfactory cues. This unit describes a method for performing STFP in mutant mice. The method explains how to address potential confounding factors and avoid possib... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927019 Wed, 01 Sep 2004 04:00:00 +0100 1927019 http://www.medworm.com/index.php?rid=1927016&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428613%26dopt%3DAbstract Authors: Kanarek RB To address the question of how animals choose diets from an array of nutritionally different foods, researchers have designed experiments in which animals are provided with separate sources of the three macronutrients, protein, fat and carbohydrate. These experiments are useful for investigating the neuroanatomical and neurochemical mechanisms involved in food choice, as well as for assessing the effects of changes in physiological status (e.g., pregnancy; lactation and increased energy expenditure) or disease states (e.g., diabetes; and obesity) on nutrient intakes. PMID: 18428613 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927016 Wed, 01 Sep 2004 04:00:00 +0100 1927016 http://www.medworm.com/index.php?rid=1927014&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428615%26dopt%3DAbstract Authors: Ling GS, Lee EY, Kalehua AN Traumatic brain injury is a leading cause of death and disability, particularly among young adults. During closed head trauma, the injury process is initiated by the impact of the brain against the inner table of the calvarium. Subsequently, there is prompt initiation of a complex biochemical, cellular, and physiological injury cascade that may take days to complete. From a functional standpoint, this culminates in neurologic dysfunction and, if severe, death. This unit describes an impact-induced brain trauma model in rats which replicates nonpenetrating head injury. It does not model either penetrating or ischemic brain injuries. PMID: 18428615 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927014 Wed, 01 Sep 2004 04:00:00 +0100 1927014 http://www.medworm.com/index.php?rid=1927035&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428594%26dopt%3DAbstract Authors: Wilson CJ, Sachdev RN Many physiological studies require microscopic examination of the recorded neuron for identification. This unit describes how intracellular and extracellular recording can be combined with single-neuron staining to enable sequential physiological and morphological studies. PMID: 18428594 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927035 Sat, 01 May 2004 04:00:00 +0100 1927035 http://www.medworm.com/index.php?rid=1927030&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428599%26dopt%3DAbstract Authors: Vicario-Abejón C In culture, hippocampal cells can develop to express neuronal antigens and acquire mature neuronal morphologies, including axons, complex dendritic trees, and synapses that are electrophysiologically active. This system is suitable for studying neuronal differentiation and other events, such as synaptogenesis. It is also a valuable model for investigating synaptic plasticity and exploring the mechanisms of neuronal degeneration. This unit provides a protocol for culturing neurons prepared from embryonic (E-18) rat or mouse hippocampus, but could also be used to grow neurons from embryonic cortex, olfactory bulb, striatum, or spinal cord. A second method is included for preparing neuronal cultures from embryos with different genotypes, such as those from t... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927030 Sat, 01 May 2004 04:00:00 +0100 1927030 http://www.medworm.com/index.php?rid=1927025&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428604%26dopt%3DAbstract Authors: Rae JL, Levis RA Patch clamping refers to a wide range of electrophysiological measurements, all of which have in common the use of patch pipets and the formation of gigaohm seals. The purpose of this unit is to describe the fabrication of patch pipets. The aspects of the pipet geometry that are important to different applications and the different procedures that have been found to most reliably and simply achieve these results are described. Parameters for glass selection are detailed in the beginning of the unit. Pulling patch and whole-cell pipets, elastomer coating, fire polishing, pipet filling, and pipet testing in an experimental setup are highlighted. Additional support protocols describe alternative ways to optimize pipet geometry and cleaning the glass before pullin... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927025 Sat, 01 May 2004 04:00:00 +0100 1927025 http://www.medworm.com/index.php?rid=1927023&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428606%26dopt%3DAbstract Authors: File SE, Lippa AS, Beer B, Lippa MT Animal tests of anxiety are used to screen novel compounds for anxiolytic or anxiogenic activity, to investigate the neurobiology of anxiety, and to assess the impact of other occurrences such as exposure to predator odors or early rearing experiences. This unit presents protocols for the most commonly used animal tests of anxiety. The Geller-Seifter conflict test, the social interaction test, light/dark exploration, the elevated plus-maze, defensive burying, and the thirsty rat conflict. The protocols are described in terms of drug screening tests, but can be modified easily for other purposes. PMID: 18428606 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927023 Sat, 01 May 2004 04:00:00 +0100 1927023 http://www.medworm.com/index.php?rid=1927022&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428607%26dopt%3DAbstract Authors: Wenk GL Behavioral tasks must be evaluated in terms of the cognitive functions they require in order to be performed. All of the tasks described in this chapter can be used with each of four experimental manipulations: stimulation of a single brain region by drugs or small electrical current, impairment of normal function by production of a lesion or administration of appropriate pharmacological agents, recording of brain activity during the performance of a specific behavioral task, or behavioral phenotyping of transgenic and knockout mice for genes expressed in specific brain regions. This unit describes protocols for the radial arm maze task and the water maze task, both of which require intact spatial memory abilities. PMID: 18428607 [PubMed - indexed for MEDLINE] (Sou... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927022 Sat, 01 May 2004 04:00:00 +0100 1927022 http://www.medworm.com/index.php?rid=1927033&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428596%26dopt%3DAbstract Authors: Volpicelli-Daley LA, Levey A The immunohistological methods described in this unit can be used to determine the precise localization of neurochemicals, receptors, and proteins throughout the nervous system. Determining the localization of a protein within defined brain nuclei and neuronal cell populations can provide important clues regarding its potential function. Immunoperoxidase reactions and light microscopy are commonly used to visualize the distribution of a single primary antibody directed to an antigen of interest. Double-labeling immunofluorescence and confocal microscopy techniques detect the localization of one protein relative to another protein and allow analysis of colocalization at a cellular and subcellular level. The colocalization of two proteins can also be... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927033 Sun, 01 Feb 2004 05:00:00 +0100 1927033 http://www.medworm.com/index.php?rid=1927032&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428597%26dopt%3DAbstract Authors: Young WS, Mezey E Expression of genes is manifested by the production of RNA transcripts within cells. Hybridization histochemistry (or in situ hybridization) permits localization of these transcripts with cellular resolution or better. Furthermore, the relative amounts of transcripts detected within different tissues or the same tissues in different states (e.g., physiological or developmental) may be quantified. This unit describes hybridization histochemical techniques using either oligonucleotide probes or RNA probes (riboprobes). Also presented is the use of probes labeled with digoxigenin for colorimetric detection of RNA transcripts and a technique to detect the Y chromosome using either mouse or human riboprobes. Finally, a procedure is presented for the autoradiograph... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927032 Sun, 01 Feb 2004 05:00:00 +0100 1927032 http://www.medworm.com/index.php?rid=1927027&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428602%26dopt%3DAbstract Authors: Swanson GT, Contractor A This unit describes techniques for the preparation of mouse cerebellar slices and electrophysiological recording from neurons in the slice. The cerebellum provides a model of motor learning that can be correlated with alterations in synaptic function. In addition, the architecture and well-defined synaptic pathways in the cerebellar slice make this preparation a useful model for exploring general principles of synaptic transmission. PMID: 18428602 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927027 Sun, 01 Feb 2004 05:00:00 +0100 1927027 http://www.medworm.com/index.php?rid=1927024&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428605%26dopt%3DAbstract Authors: Heyser CJ Developmental tests are used to characterize early markers of behavior for investigation of the neurobiology of these behaviors, and to assess the impact of early prenatal or postnatal insult. These perturbations may include pharmacological, environmental, and genetic manipulations. At birth the rat is capable of some specific activities, but its movements are uncoordinated and seemingly random, its tactile sensitivity is not fully developed, and its ear canals and eyes remain closed until several days after birth. Postnatal development consists mainly of the continuation of processes begun earlier. This unit presents protocols for the most commonly used animal tests of developmental reflexology, including negative geotaxis, cliff avoidance, placing responses, tactil... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927024 Sun, 01 Feb 2004 05:00:00 +0100 1927024 http://www.medworm.com/index.php?rid=1927020&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428609%26dopt%3DAbstract Authors: Barrett JE, Vanover KE This unit describes the use of an automated procedure for developing an operant response ("autoshaping") in the mouse. The method has applications in the study of the acquisition of behavior (learning) as well as for the assessment of memory or retention of that task. PMID: 18428609 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927020 Sun, 01 Feb 2004 05:00:00 +0100 1927020 http://www.medworm.com/index.php?rid=1927045&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428584%26dopt%3DAbstract Authors: Geyer MA, Dulawa SC The study of the startle response and its plasticity provides a powerful research paradigm for examining the neural control of behavior. The neural substrates underlying phenomena such as information-processing functions, including sensorimotor gating, affective modulation, and habituation, can be studied using startle plasticity as an operational measure. The mouse is rapidly becoming the most powerful model system for the study of human CNS function and is highly amenable to both genetic and pharmacological study. This unit describes procedures for the assessment of the startle response to acoustic stimuli in mice; methods for assessing prepulse inhibition (PPI) of startle, with treatments including drug administration and genetic manipulation; and method... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927045 Sat, 01 Nov 2003 05:00:00 +0100 1927045 http://www.medworm.com/index.php?rid=1927039&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428590%26dopt%3DAbstract Authors: Sajdyk TJ, Keim SR, Thielen SR, Fitz SD, Shekhar A This unit describes a putative animal model for panic disorder. The basic premise is that pharmacological disruption of critical brain regions implicated in the circuitry of anxiety will lead to a condition similar to that of the human disorder. A clinically relevant test, the sodium lactate challenge, is utilized to assess parallels between the human condition and this rat model. PMID: 18428590 [PubMed - indexed for MEDLINE] (Source: Current Protocols in Neuroscience) Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927039 Sat, 01 Nov 2003 05:00:00 +0100 1927039 http://www.medworm.com/index.php?rid=1927040&cid=s_38151_168_f&fid=38151&url=http%3A%2F%2Fwww.ncbi.nlm.nih.gov%2Fentrez%2Fquery.fcgi%3Ftmpl%3DNoSidebarfile%26db%3DPubMed%26cmd%3DRetrieve%26list_uids%3D18428589%26dopt%3DAbstract Authors: Bergeron M In the rat photochemical cortical lesion model described in this unit, an intravascular photochemical reaction induces endothelial damage resulting in platelet aggregation, thrombosis, thrombotic response (secretion of factors by the platelets) and permanent cerebral vascular occlusion. Because thrombosis is produced in pial vessels, the resulting cortical infarct is generally smaller and more reproducible than in the models involving occlusion of the middle cerebral artery. The surgical procedures involved are limited, making this model generally easier to perform and less invasive than most other models of permanent focal ischemia that involve mechanical occlusion of major cerebral arteries. PMID: 18428589 [PubMed - indexed for MEDLINE] (Source: Current Protoc... Current Protocols in Neuroscience journals http://www.medworm.com/rss/comments.php?id=1927040 Tue, 01 Jul 2003 04:00:00 +0100 1927040