Immunohistochemical detection of raf protein kinase in cerebral cortical areas of adult guinea pigs and rats

Brain Res. 1993 Nov 12;627(2):225-38. doi: 10.1016/0006-8993(93)90325-h.

Abstract

The raf protooncogenes are the cellular counterparts of the v-raf oncogene expressed by a murine sarcoma virus. The raf protooncogenes encode cytoplasmic serine/threonine-specific protein kinases which can be activated from different growth factor receptors by phosphorylation. The mRNAs of raf protooncogenes are found in a large variety of normal adult tissues, including the central nervous system. As concerns the distribution and localization of their protein products (the raf kinases), very few data are to be found in the literature. This is the first detailed description of their light microscopic localization in neocortical and allocortical areas of rodents. Preembedding immunohistochemical studies were performed on vibratome sections from the brains of adult guinea pigs and albino rats. The localizations of two isoenzymes, raf-1 kinase and B-raf kinase, were studied with the help of isoenzyme-specific polyclonal antibodies. Both of the antibodies detected raf protein-like immunoreactivity in many neurons and scattered glial cells of the sensory neocortex, and the cingular, pyriform, perirhinal and entorhinal allocortical areas. Pyramidal and non-pyramidal cells of Ammon's horn, granule cells of the dentate fascia and the large neurons in the hilar region were immunoreactive, too. The findings indicated that B-raf protein kinase and raf-1 kinase are present almost ubiquitously in the neurons of the investigated cortical structures. The intensity of staining obtained with serial dilutions of the antibodies indicated that the cytoplasmic concentration of B-raf kinase is tended to be higher than that of raf-1 kinase. The present findings suggested that the raf kinases are localized in postsynaptic structures, mainly in dendrites and cell bodies. Their cytosolic localization and their ability to undergo intracellular translocation during activation and phosphorylation raise the possibility that they play a pivotal role in the intracellular signaling of neurons.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal
  • Antibody Specificity
  • Auditory Cortex / enzymology
  • Cerebral Cortex / enzymology*
  • Guinea Pigs
  • Immunohistochemistry
  • Isoenzymes / analysis*
  • Limbic System / enzymology
  • Male
  • Molecular Sequence Data
  • Protein Serine-Threonine Kinases / analysis*
  • Proto-Oncogene Proteins / analysis*
  • Proto-Oncogene Proteins c-raf
  • Rats
  • Rats, Wistar
  • Sexual Maturation
  • Somatosensory Cortex / enzymology

Substances

  • Antibodies, Monoclonal
  • Isoenzymes
  • Proto-Oncogene Proteins
  • Protein Serine-Threonine Kinases
  • Proto-Oncogene Proteins c-raf