Full Description
Neurobiologists are bound to differ in their perceptions of what the discipline of molecular neurobiology should encompass. We have taken the view that molecular neurobiology should cover any aspect of brain science that uses the techniques of modern molecular biology, though we accept the fact that classification of a technique as a "biochemical* or "molecular biological* one is in itself somewhat arbitrary. Each chapter of this volume sets out to identify a clear problem in neurobiology and to place it in its context within the literatu- i.e., indicating how the solution of the problem will advance knowledge in the field. The core of the chapter then details the approaches taken to solve the problem, in sufficient detail that the reader can appre- ate exactly why a specific strategy was adopted and how it was executed. Each chapter also includes detailed protocols providing all the inf- mation necessary to reproduce the technique and its results in any appropriately equipped laboratory.
Contents
Separation of Large DNA Molecules by Pulsed-Field Gel Electrophoresis.- The Isolation of Genomic DNA from Invertebrates.- Analysis of Mitochondrial DNA Mutations.- The Design and Use of Oligonucleotides.- The Use of Degenerate Oligonucleotides for Polymerase Chain-Reaction-Based Isolation of Related DNA Sequences.- Isolation of Photoreceptor Cell-Specific MEKA cDNA by Differential Hybridization.- Generation of Isoform-Specific Antisera from Cloned cDNAs.- Determination of Transcriptional Initiation Sites and Their Usage in the Nervous System.- Mapping of Trans-Acting Factor Binding in the Nervous System.- In Situ Hybridization to Brain Tissue Sections Using Labeled Single-Strand Complementary RNA Probes.- In Situ Hybridization Histochemistry Using Alkaline Phosphatase-Labeled Oligodeoxynucleotide Probe.- The Identification of Neuropeptide Gene Regulatory Elements in Transgenic Mice.- Expression of Exogenous Ion Channels and Neurotransmitter Receptors in RNA-Injected Xenopus Oocytes.- Analysis of Insulin and Insulin-Like Growth Factor-I Receptors in Neural Tissues.- Use of Affinity Chromatography in Purification of A1 Adenosine Receptors from Rat Brain Membranes.- Purification and Structure of L-Type Calcium Channels.- Purification and Reconstitution of the Ryanodine-Sensitive Ca2+ Release Channel Complex from Muscle Sarcoplasmic Reticulum.- Identification of a Ligand-Gated Ion Channel by Photoaffinity Labeling and Microsequencing.- Voltage-Gated Ion Channels.- An Electrophysiological Approach to the Regulation of Neuronal Voltage-Activated Calcium Channels by Guanine Nucleotide Binding Proteins.
