In Situ Hybridization in Neurobiology: Advances in MethodologyJames H. Eberwine, Karen L. Valentino, Jack D. Barchas Oxford University Press, 1994 - 199 pages In situ hybridization has become an important and widely used research tool for neurobiologists since the authors' first book on the subject, In Situ Hybridization: Applications to Neurobiology (Oxford, 1987). This new volume examines the methodological advances that have been made in in situ hybridization techniques and their impact on the study of the nervous system. These include methods for improving sensitivity, ways of examining multiple mRNAs within the same section, ways of examining both proteins and mRNAs within the same section, and methods for using in situ hybridization technology to examine areas of cellular biological significance, such as translational control. The development of alternative labels to radioactivity for the detection of hybridized probes, and the combined use of in situ hybridization and immunocytochemistry is also covered in this book. It discusses the quantification of in situ hybridization signal and presents an overview of information generated by using in situ hybridization in diagnostic medicine. This volume will be essential to researchers in the neurosciences as well as those in other disciplines. |
Contents
Methodological Considerations in the Utilization of In Situ | 3 |
Combining Nonisotopic In Situ Hybridization Histochemistry with Steroid | 24 |
Development of Techniques to Combine Isotopic and Nonisotopic In Situ | 43 |
Enzyme Histochemical Detection of Neuronal mRNA | 63 |
Characterization of the Receptor Complement of Individual Neurons | 78 |
Assessment of Peptide Secretion and Gene Expression Within Individual | 96 |
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30 minutes amplification analysis antibody aRNA autoradiography Baldino binding biotin biotinylated bridization cDNA cDNA synthesis cellular Chesselet clones colocalization concentration containing coverslips cRNA probes cytoplasmic denatured DEPC DEPC-treated detection digoxigenin digoxigenin-labeled DNA polymerase dNTPs dual-label ISHH duplexes Eberwine Emson emulsion encoding enzyme estrogen receptor ethanol exposure Figure fixed formamide GABA receptor gene expression Haase histochemistry hormone hybridization histochemistry hybridization probes hybridization signal incubation infection IS-PCR Kiyama levels of labeling LHRH LHRH mRNA LHRH neurons method molecular mRNA mRNA levels NaCl neurons Neuroscience nonisotopic nonradioactive nonspecific Northern blots nucleic acid nucleotides oligonucleotide probes paraformaldehyde peptide pituitary polymerase chain reaction POMC prehybridization primary transcript primer procedure proopiomelanocortin protein quantitative radiolabeled receptor subunit region reverse transcriptase riboprobes RNase room temperature sensitivity sequence silver grains situ hybridization slides sodium solution staining stem-loop steroid studies synthetic oligonucleotide technique tissue sections Toran-Allerand washed µg/ml