In Situ Hybridization for Peptide mRNA

  • Giorgio Terenghi
  • Julia M. Polak

Abstract

The introduction of in situ hybridization for mRNA using labelled complementary nucleic acid probes has made possible the understanding of gene expression at the cellular level. Different types of probe have been used for in situ hybridization, including cDNA sequences, synthetic oligonucleotides, and complementary RNA (cRNA) probes. Hybridization can be carried out on sections of fixed tissue as well as on cell culture preparations. The approach to radioactive and non-radioactive in situ hybridization is similar, with some variations in the concentration of the solutions or the timing of different steps. The choice of radiolabel is generally determined by a balance between speed and resolution, and both 32P and 35S are suited for studies of the neuroendocrine system. The use of digoxigenin is recommended when working with non-radioactive probes, as it sometimes gives lower background than biotin. The detection of either non-isotopic reporter molecule can be carried out using a wide range of immunohisto-chemical methods. We have applied in situ hybridization for various studies of regulatory peptides, such as anatomical identification of peptide mRNA, detection of functional changes of peptide synthesis and secretion in different endocrine situations. In situ hybridization can offer a suitable complement or alternative to morphological investigation at light microscopical level by immunohistochemistry.

Keywords

Dust Lymphoma Adenocarcinoma Polypeptide Dehydration 

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Copyright information

© Springer Science+Business Media New York 1994

Authors and Affiliations

  • Giorgio Terenghi
    • 1
  • Julia M. Polak
    • 1
    • 2
  1. 1.Department of HistochemistryRoyal Postgraduate Medical School Hammersmith HospitalLondonUK
  2. 2.Department of HistochemistryRoyal Postgraduate Medical School Hammersmith HospitalLondonUK

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