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Assessment of Parkinson disease manifestations.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 ratin...
Source: Current Protocols in Neuroscience - September 30, 2009 Category: Neuroscience Authors: Perlmutter JS Tags: Curr Protoc Neurosci Source Type: journals

EEG recording and analysis for sleep research.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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)
Source: Current Protocols in Neuroscience - September 30, 2009 Category: Neuroscience Authors: Campbell IG Tags: Curr Protoc Neurosci Source Type: journals

Neuropsychological assessment in the diagnosis of Alzheimer's disease.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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)
Source: Current Protocols in Neuroscience - September 30, 2009 Category: Neuroscience Authors: Baldwin S, Farias ST Tags: Curr Protoc Neurosci Source Type: journals

Imaging nervous system activity.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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)
Source: Current Protocols in Neuroscience - September 30, 2009 Category: Neuroscience Authors: Fields DR, Shneider N, Mentis GZ, O'Donovan MJ Tags: Curr Protoc Neurosci Source Type: journals

EEG recording in rodents, with a focus on epilepsy.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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)
Source: Current Protocols in Neuroscience - September 30, 2009 Category: Neuroscience Authors: Martín Del Campo C, Velázquez JL, Freire MA Tags: Curr Protoc Neurosci Source Type: journals

Recapitulation and Reversal of a Persistent Depression-like Syndrome in Rodents.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 C...
Source: Current Protocols in Neuroscience - September 30, 2009 Category: Neuroscience Authors: Gourley SL, Taylor JR Tags: Curr Protoc Neurosci Source Type: journals

Assessing Reproductive Status/Stages in Mice.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 ty...
Source: Current Protocols in Neuroscience - June 30, 2009 Category: Neuroscience Authors: Caligioni CS Tags: Curr Protoc Neurosci Source Type: journals

Identification of Neural Programmed Cell Death Through the Detection of DNA Fragmentation In Situ and by PCR.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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)
Source: Current Protocols in Neuroscience - June 30, 2009 Category: Neuroscience Authors: Yung YC, Kennedy G, Chun J Tags: Curr Protoc Neurosci Source Type: journals

Production of Polyclonal Antisera.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 ...
Source: Current Protocols in Neuroscience - June 30, 2009 Category: Neuroscience Authors: Cooper HM, Paterson Y Tags: Curr Protoc Neurosci Source Type: journals

Measurement of Oxygen Radicals and Lipid Peroxidation in Neural Tissues.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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, t...
Source: Current Protocols in Neuroscience - June 30, 2009 Category: Neuroscience Authors: Hall ED, Bosken JM Tags: Curr Protoc Neurosci Source Type: journals

Detection and Quantification of Neurotransmitters in Dialysates.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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] (Sour...
Source: Current Protocols in Neuroscience - June 30, 2009 Category: Neuroscience Authors: Zapata A, Chefer VI, Shippenberg TS, Denoroy L Tags: Curr Protoc Neurosci Source Type: journals

Simple Behavioral Assessment of Mouse Olfaction.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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. Pr...
Source: Current Protocols in Neuroscience - June 30, 2009 Category: Neuroscience Authors: Yang M, Crawley JN Tags: Curr Protoc Neurosci Source Type: journals

Differentiation of embryonic stem cells.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 embr...
Source: Current Protocols in Neuroscience - March 31, 2009 Category: Neuroscience Authors: Mansouri A, Fukumitsu H, Schindehuette J, Krieglstein K Tags: Curr Protoc Neurosci Source Type: journals

Transfection of cultured primary neurons via nucleofection.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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. ...
Source: Current Protocols in Neuroscience - March 31, 2009 Category: Neuroscience Authors: Zeitelhofer M, Vessey JP, Thomas S, Kiebler M, Dahm R Tags: Curr Protoc Neurosci Source Type: journals

Overview of brain microdialysis.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 ava...
Source: Current Protocols in Neuroscience - March 31, 2009 Category: Neuroscience Authors: Chefer VI, Thompson AC, Zapata A, Shippenberg TS Tags: Curr Protoc Neurosci Source Type: journals

Microdialysis in rodents.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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)
Source: Current Protocols in Neuroscience - March 31, 2009 Category: Neuroscience Authors: Zapata A, Chefer VI, Shippenberg TS Tags: Curr Protoc Neurosci Source Type: journals

Fear extinction in rodents.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 labo...
Source: Current Protocols in Neuroscience - March 31, 2009 Category: Neuroscience Authors: Chang CH, Knapska E, Orsini CA, Rabinak CA, Zimmerman JM, Maren S Tags: Curr Protoc Neurosci Source Type: journals

Evolution of neurotoxins: from research modalities to clinical realities.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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. Add...
Source: Current Protocols in Neuroscience - January 1, 2009 Category: Neuroscience Authors: Kostrzewa RM Tags: Curr Protoc Neurosci Source Type: journals

Identifying novel protein-protein interactions using co-immunoprecipitation and mass spectroscopy.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 i...
Source: Current Protocols in Neuroscience - January 1, 2009 Category: Neuroscience Authors: Free RB, Hazelwood LA, Sibley DR Tags: Curr Protoc Neurosci Source Type: journals

An animal model of premenstrual dysphoric disorder sensitive to antidepressants.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 bu...
Source: Current Protocols in Neuroscience - January 1, 2009 Category: Neuroscience Authors: Schneider T, Popik P Tags: Curr Protoc Neurosci Source Type: journals

Culture of postnatal mesencephalic dopamine neurons on an astrocyte monolayer.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 develo...
Source: Current Protocols in Neuroscience - July 1, 2008 Category: Neuroscience Authors: Fasano C, Thibault D, Trudeau LE Tags: Curr Protoc Neurosci Source Type: journals

Conditional gene expression and targeting in neuroscience research.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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)
Source: Current Protocols in Neuroscience - July 1, 2008 Category: Neuroscience Authors: Morozov A Tags: Curr Protoc Neurosci Source Type: journals

Assessment of cell viability in primary neuronal cultures.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 viab...
Source: Current Protocols in Neuroscience - July 1, 2008 Category: Neuroscience Authors: Aras MA, Hartnett KA, Aizenman E Tags: Curr Protoc Neurosci Source Type: journals

The Majchrowicz binge alcohol protocol: an intubation technique to study alcohol dependence in rats.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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. Aft...
Source: Current Protocols in Neuroscience - July 1, 2008 Category: Neuroscience Authors: Faingold CL Tags: Curr Protoc Neurosci Source Type: journals

Vapor inhalation of alcohol in rats.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 alc...
Source: Current Protocols in Neuroscience - July 1, 2008 Category: Neuroscience Authors: Gilpin NW, Richardson HN, Cole M, Koob GF Tags: Curr Protoc Neurosci Source Type: journals

Retrograde axonal tracing with fluorescent markers.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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, incl...
Source: Current Protocols in Neuroscience - April 1, 2008 Category: Neuroscience Authors: Schofield BR Tags: Curr Protoc Neurosci Source Type: journals

Ballistic labeling with fluorescent dyes and indicators.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 r...
Source: Current Protocols in Neuroscience - April 1, 2008 Category: Neuroscience Authors: Morgan JL, Wong RO Tags: Curr Protoc Neurosci Source Type: journals

Use of FM1-43 and other derivatives to investigate neuronal function.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 synapti...
Source: Current Protocols in Neuroscience - April 1, 2008 Category: Neuroscience Authors: Cousin MA Tags: Curr Protoc Neurosci Source Type: journals

Dissection and culture of embryonic spinal commissural neurons.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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,...
Source: Current Protocols in Neuroscience - April 1, 2008 Category: Neuroscience Authors: Moore SW, Kennedy TE Tags: Curr Protoc Neurosci Source Type: journals

Schedule-induced polydipsia: a rat model of obsessive-compulsive disorder.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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-induc...
Source: Current Protocols in Neuroscience - April 1, 2008 Category: Neuroscience Authors: Platt B, Beyer CE, Schechter LE, Rosenzweig-Lipson S Tags: Curr Protoc Neurosci Source Type: journals

Mouse and rat anesthesia and analgesia.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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)
Source: Current Protocols in Neuroscience - January 1, 2008 Category: Neuroscience Authors: Davis JA Tags: Curr Protoc Neurosci Source Type: journals

Biochemical fractionation of brain tissue for studies of receptor distribution and trafficking.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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...
Source: Current Protocols in Neuroscience - January 1, 2008 Category: Neuroscience Authors: Hallett PJ, Collins TL, Standaert DG, Dunah AW Tags: Curr Protoc Neurosci Source Type: journals

Neural stem cell transplantation in mouse brain.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 ...
Source: Current Protocols in Neuroscience - January 1, 2008 Category: Neuroscience Authors: Lee JP, McKercher S, Muller FJ, Snyder EY Tags: Curr Protoc Neurosci Source Type: journals

Overview of mouse assays of ethanol intoxication.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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...
Source: Current Protocols in Neuroscience - January 1, 2008 Category: Neuroscience Authors: Crabbe JC, Cameron AJ, Munn E, Bunning M, Wahlsten D Tags: Curr Protoc Neurosci Source Type: journals

Alphaviruses: Semliki Forest virus and Sindbis virus vectors for gene transfer into neurons.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 develope...
Source: Current Protocols in Neuroscience - October 1, 2007 Category: Neuroscience Authors: Ehrengruber MU, Lundstrom K Tags: Curr Protoc Neurosci Source Type: journals

Split-ubiquitin system for identifying protein-protein interactions in membrane and full-length proteins.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 screeni...
Source: Current Protocols in Neuroscience - October 1, 2007 Category: Neuroscience Authors: Grefen C, Lalonde S, Obrdlik P Tags: Curr Protoc Neurosci Source Type: journals

Models of nociception: hot-plate, tail-flick, and formalin tests in rodents.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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] (Sourc...
Source: Current Protocols in Neuroscience - October 1, 2007 Category: Neuroscience Authors: Bannon AW, Malmberg AB Tags: Curr Protoc Neurosci Source Type: journals

Ratings of L-DOPA-induced dyskinesia in the unilateral 6-OHDA lesion model of Parkinson's disease in rats and mice.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 condition...
Source: Current Protocols in Neuroscience - October 1, 2007 Category: Neuroscience Authors: Cenci MA, Lundblad M Tags: Curr Protoc Neurosci Source Type: journals

Gene delivery using helper virus-free HSV-1 amplicon vectors.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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...
Source: Current Protocols in Neuroscience - July 1, 2007 Category: Neuroscience Authors: Fraefel C Tags: Curr Protoc Neurosci Source Type: journals

Overview of gene targeting by homologous recombination.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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, crite...
Source: Current Protocols in Neuroscience - July 1, 2007 Category: Neuroscience Authors: Mortensen R Tags: Curr Protoc Neurosci Source Type: journals

Production of antisera using fusion proteins.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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...
Source: Current Protocols in Neuroscience - July 1, 2007 Category: Neuroscience Authors: Dodson SE, Heilman CJ, Kahn RA, Levey AI Tags: Curr Protoc Neurosci Source Type: journals

Locomotor behavior.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 locomot...
Source: Current Protocols in Neuroscience - July 1, 2007 Category: Neuroscience Authors: Pierce RC, Kalivas PW Tags: Curr Protoc Neurosci Source Type: journals

Histochemical methods for the detection of apoptosis in the nervous system.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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...
Source: Current Protocols in Neuroscience - April 1, 2007 Category: Neuroscience Authors: Oo TF, Burke RE Tags: Curr Protoc Neurosci Source Type: journals

Lentiviral strategies for RNAi knockdown of neuronal genes.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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. Lenti...
Source: Current Protocols in Neuroscience - April 1, 2007 Category: Neuroscience Authors: Crittenden JR, Heidersbach A, McManus MT Tags: Curr Protoc Neurosci Source Type: journals

Delay discounting and impulsive choice in the rat.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 de...
Source: Current Protocols in Neuroscience - April 1, 2007 Category: Neuroscience Authors: Mar AC, Robbins TW Tags: Curr Protoc Neurosci Source Type: journals

Overview of animal models of schizophrenia.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 wit...
Source: Current Protocols in Neuroscience - April 1, 2007 Category: Neuroscience Authors: Powell SB, Geyer MA Tags: Curr Protoc Neurosci Source Type: journals

Techniques for mammalian cell tissue culture.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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)
Source: Current Protocols in Neuroscience - January 1, 2007 Category: Neuroscience Authors: Phelan MC Tags: Curr Protoc Neurosci Source Type: journals

Whole-cell recording in vivo.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 t...
Source: Current Protocols in Neuroscience - January 1, 2007 Category: Neuroscience Authors: DeWeese MR Tags: Curr Protoc Neurosci Source Type: journals

Electrophysiological recordings from neonatal neocortical brain slices.Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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)
Source: Current Protocols in Neuroscience - January 1, 2007 Category: Neuroscience Authors: Daw M, Isaac J Tags: Curr Protoc Neurosci Source Type: journals

Assays for cyclic nucleotide-specific phosphodiesterases (PDEs) in the central nervous system (PDE1, PDE2, PDE4, and PDE10).Email this article to a colleague. Save this article to My Clippings. Discuss or comment on this article.
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 rel...
Source: Current Protocols in Neuroscience - January 1, 2007 Category: Neuroscience Authors: Deng C, Wang D, Bugaj-Gaweda B, De Vivo M Tags: Curr Protoc Neurosci Source Type: journals