Abstract
Publisher Summary In situ hybridization histochemistry (ISHH) can be used to detect the transcribed messenger RNA (mRNAs) coding for neuropeptides (or for enzymes or other proteins) in cells that express the corresponding genes. Tissue sections are incubated with labeled complementary DNA (cDNA) or complementary (c) RNA probes that form hybrids with the cellular mRNA, and the location of labeled cells is then determined usually by autoradiography for radioactively tagged probes. The probes can be designed to distinguish among a gene's splicing alternatives or among related genes' mRNAs. They also readily penetrate processed tissue sections and yield good signal-to-noise ratios without resorting to nucleases to reduce background. The use of ribonucleases to treat sections prior to hybridization is similar to treating sections with proteases prior to immunohistochemistry and is of questionable value. Similarly, adding excess unlabeled probe to the hybridization simply reduces the specific activity of the probe.
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