Histochemistry

, Volume 94, Issue 2, pp 187–190 | Cite as

In situ hybridization of semithin Epon sections with BrdU labelled oligonucleotide probes

  • G. F. Jirikowski
  • J. F. Ramalho-Ortigao
  • K. W. Kesse
  • F. E. Bloom
Article

Summary

We recently described a nonradioactive method for in situ hybridization with 5-bromo-2-deoxyuridine (BrdU) labelled oligonucleotide probes. An antibody to BrdU and immunocytochemistry were used in order to detect the hybridization signal. We have now applied this method to semithin Epon sections, in order to hybridize consecutive sections through single cells with different probes and to stain them with antibodies to neuropeptides. It could be shown that Epon embedding preserves mRNA well. In the present study we used a BrdU labelled synthetic oligonucleotide probe complementary to a fragment of the vasopressin precursor and an antibody to Arg-vasopressin. Vasopressin mRNA was demonstrable in a fraction of the vasopressin immunoreactive neurons in the magnocellular nuclei. In addition some of the magnocellular neurons showed either hybridization or vasopressin immunostaining only, perhaps indicating different stages of synthetic and secretory activity.

The method described seems to be a valuable tool for studying synthetic activity in peptidergic neurons on a single cell level. The method might also have potential for in situ hybridization on the electronmicroscopical level.

Keywords

Vasopressin Oxytocin Oligonucleotide Probe Consecutive Section Magnocellular Neuron 

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Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • G. F. Jirikowski
    • 1
    • 2
    • 3
  • J. F. Ramalho-Ortigao
    • 2
    • 3
  • K. W. Kesse
    • 4
  • F. E. Bloom
    • 1
  1. 1.Department of NeuropharmacologyScripps Clinic and Research FoundationUlmFederal Republic of Germany
  2. 2.Abteilung Anatomie und Zellbiologie and Sektion PolymereUniversity of UlmUlmFederal Republic of Germany
  3. 3.Abteilung AnatomieUniversity of UlmUlmFederal Republic of Germany
  4. 4.Department of AnatomyUniversity of AkraAkraGhana

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