Short Protocols in Neuroscience is an easy-to-use compendium of cellular and molecular methods. It provides a portable and streamlined resource, covering:

• methods for analysis of neural cell structure and function
• isolation and culture of neural cells
• characterization of neural cell gene expression
• and more

Contents

Neuroanatomical Methods

• Basic Neuroanatomical Methods
• Immunohistochemical Localization of Proteins in the Nervous System
• Hybridization Histochemistry of Neural Transcripts
• Receptor Binding Techniques
• Intracellular and Juxtacellular Staining with Biocytin

Imaging

• Theory and Application of Fluorescence Microscopy
• Basic Confocal Microscopy
• Imaging Nervous System Activity
• Loading Neurons with Dextran-Conjugated Calcium Indicators in Intact Nervous Tissue
• Using FM1-43 and Other Derivatives to Investigate Neuronal Function
• Multidisciplinary Approaches to Characterizing Synaptic Vesicle Proteins

Cellular and Developmental Neuroscience

• Culture of Neuroepithelial Stem Cells
• Differentiation of Embryonic Stem Cells
• Quantitative Analysis of In Vivo Cell Proliferation
• Long-Term Culture of Hippocampal Neurons
• Culture of Substantia Nigra Neurons
• Isolation and Purification of Primary Rodent Astrocytes
• Experimental Transplantation in the Embryonic, Neonatal, and Adult Mammalian Brain

Gene Cloning, Expression, Metagenesis

• PCR Cloning of Neural Gene Products
• Interaction Trap/Two Hybrid System to Identify Interacting Proteins
• cDNA Library Construction from Single Cells
• Overview of Neural Gene Expression
• Selection of Transfected Mammalian Cells
• Protein Expression in the Drosophila Schneider 2 Cell System
• Protein Expression in the Baculovirus System
• Gene Transfer into Neural Cells in Vitro Using Adenoviral Vectors
• Gene Transfer into Rat Brain Using Adenoviral Vectors
• Production of High-Titer Lentivirus Vectors
• Production of Recombinant Adeno-Associated Viral Vectors and Use In Vitro and In Vivo Administration
• Alphavirus-Mediated Gene Transfer into Neurons
• Overview of Gene Delivery into Cells Using HSV-1 Vectors
• Generation of High-Titer Defective HSV-1 Vectors
• Gene Delivery Using Helper Virus–Free HSV-1 Amplicon Vectors
• Overview of Nucleic Acid Arrays
• Preparation of mRNA for Expression Monitoring
• Gene Expression Analysis Using cDNA Microarrays
• Overview of Gene Targeting by Homologous Recombination

Molecular Neuroscience

• Analysis of RNA in Northern and Slit-Blot Hybridization
• RNA Analysis by Nuclease Protection
• Analysis of mRNA Populations from Single Live and Fixed Cells from the Central Nervous system
• Overview of RNAi
• Production of Antipeptide Antisera
• Production of Antisera Using Fusion Proteins
• Immunoblotting and Immunodetection
• The Use of Phage Display in Neurobiology
• Using Phage Display in Neurobiology
• Detection of Protein Phosphorylation in Tissues and Cells
• Overview of Membrane Protein Solubilization
• Immunoprecipitation
• Coimmunoprecipitation
• FRET
• An Overview on the Generation of BAC Transgenic Mice for Neuroscience Research
• Modification of Bacterial Artificial Chromosomes (BACs) and Preparation of Intact BAC DNA for Generation of Transgenic Mice

Index

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