Electron Microscopy and In Situ Hybridization

Expression of P2Y2 Receptor mRNA in the Cerebellum
  • Andrzej Loesch
  • Rainer Glass
Part of the Methods in Molecular Biology™ book series (MIMB, volume 326)

Summary

This chapter describes a pre-embedding in situ hybridization method utilizing an immunogold-silver intensification step to identify P2Y2 receptor mRNA transcripts in the adult rat cerebellum. The method was applied for ultrastructural (electron microscopic) examination. Transcripts for P2Y2 receptors, marked by the gold-silver grains, were revealed in Purkinje cells. Transcripts were essentially localized in the cytoplasm although they also appeared to be specifically associated with granular endoplasmic reticulum. This suggests that Purkinje cells may produce functional P2Y2 receptors involved in the ATP-related regulatory role in the cerebellum. The in situ hybridization method that was applied enabled simultaneous preservation of tissue ultrastructure and localization of labeled mRNA transcripts. This method may be adapted and used to study various tissues.

Key Words

In situ hybridization ultrastructure cerebellum P2Y2 receptor 

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Copyright information

© Humana Press Inc. 2006

Authors and Affiliations

  • Andrzej Loesch
  • Rainer Glass

There are no affiliations available

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