The explosion of interest in specific molecules important for brain function and dysfunction has drawn individuals from diverse backgrounds toward the use of in situ hybridization techniques. Study of the brain demands the anatomic precision and biochemical specificity that this approach can potentially bring. Workers with backgrounds in peptide neuroanatomy, neuropharmacology, molecular biology, neurovirology, neuropathology, and neurophysiology have joined together in this volume to discuss their initial experiences in applying ill situ hybridization techniques to the study of the brain. The work, although still in an early phase of development, is worthy of initial summary and dissemination. In the area of neuropeptide gene expression alone, investigators represented here describe studies of vasopressin, opiate peptides, oxytocin, vasoactive intestinal peptide, cholecystokinin, and somatostatin. Other contributions provide insight into applications of the technique to studies of the expression of genes for neurotransmitter synthesizing enzymes, viral-encoded genes, trophic factor genes, and the genes selected on the basis of their special roles in the brain. The authors provide an important series of technical perspectives, and describe specific experimental protocols. This volume should be of interest to individuals seeking an introduction to these methods, as well to those desiring an up to date precis of work in this burgeoning area. Dr. Uhl, with the sponsorship of the Howard Hughes Medical Institute, has done a superb job of assembling the leaders in this area, and in organizing the presen ta tion of their perspecti ves herein. Joseph B. Martin, M.D., Ph.D.
|Edition description:||Softcover reprint of the original 1st ed. 1986|
|Product dimensions:||7.01(w) x 10.00(h) x 0.03(d)|
Table of ContentsSection I: In Situ Hybridization: Approaches and Applications.- 1. Cellular Location and Regulation of Proenkephalin mRNA in Rat Brain.- 2. Vasopressin and Somatostatin mRNA In Situ Hybridization.- 3. Opioid Peptides and Vasopressin: The Application of In Situ Hybridization to Studies of the Hypothalamus and Pituitary.- 4. Detection of Neuropeptide mRNAs by In SituHybridization Histochemistry.- 5. In Situ Hybridization as a Quantitative Autoradiographic Method: Vasopressin and Oxytocin Gene Transcription in the Brattleboro Rat.- 6. Glucocorticoid Regulation of Vasopressin Messenger RNA.- 7. Neuro Anatomical and Developmental Expression of Novel Brain mRNAs Determined by In Situ Hybridization.- 8. In Situ Detection of GAD mRNA in Mouse Brain.- 9. The Cellular Localization of Preprotachykinin, Preproenkephalin A and Beta Prepronerve Growth Factor mRNA.- Discussion: Nerve Growth Factor Gene Expression in Mouse Hippocampal Formation.- 10. Molecular Approaches to Human Neurological Diseases and Their Animal Models.- 11. In Situ Hybridization of Somatostatin and Vasoactive Intestinal Peptide mRNA in the Rat Nervous System: Contrasting Patterns of Ontogeny.- 12. Use of In Situ Hybridization Histochemistry to Analyze Gene Transcription in Individual Cells.- 13. In Situ Hybridization for Detection of Viral Nucleic Acid in Cell Cultures and Tissues.- Discussion: Detection of Viral Nucleic Acids in the Central Nervous System.- Section II: Methodological Issues.- 14. Probes.- (A) General Features.- (B) Nick-Translation.- (C) Single-Stranded End-Labeled DNA Probes.- (1)W. Scott Young, III.- (2)Leonard G. Davis, Michael E. Lewis and Frank Baldino, Jr..- (D) Single-Stranded Multiply-Labeled cDNA Probes.- (1) Syn Probes.- (2) Single-Stranded DNA Probes Using an M13 Template.- (E) Single-Stranded Multiply-Labeled RNA Probes.- (F) Biotin Labeling.- 15. Hybridization and Wash Conditions.- 16. Quantitative In Situ Hybridization and Determination of mRNA Content.- (1)W. Scott Young, III.- (2)Michael J. Kuhar.- 17. In Situ Hybridization Histochemistry as a Supplement to Immunohistochemistry.- 18. Determination of Specificity in In Situ Hybridization.- Section III: Appendix.- Index and Glossary.