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Journal of Neuroscience, Vol 15, 2906-2919, Copyright © 1995 by Society for Neuroscience
Cellular localization of thrombin receptor mRNA in rat brain: expression by mesencephalic dopaminergic neurons and codistribution with prothrombin mRNA
JR Weinstein, SJ Gold, DD Cunningham and CM Gall
Department of Microbiology and Molecular Genetics, University of California, Irvine 92717, USA.
Cell culture studies demonstrating that the serine protease thrombin can
induce neuronal and glial process retraction, glial proliferation, and
changes in gene expression suggest a role for thrombin in CNS development,
plasticity, and response to injury. Most cellular responses to thrombin are
mediated by proteolytic activation of the cloned thrombin receptor (TR), a
member of the seven transmembrane domain, G-protein-coupled receptor
superfamily. As a step toward understanding the role of thrombin and its
receptor in the CNS, Northern blot, in situ hybridization, and
immunohistochemical techniques were used to analyze the cellular
localization of TR mRNA in weanling-age rat brain. TR mRNA was broadly
distributed across the neuraxis, although expression was very focal and
often anatomically limited within specific neural structures. The greatest
hybridization was associated with individual neurons in neocortex,
cingulate/retrosplenial cortex, and subiculum, subsets of nuclei in
hypothalamus, thalamus, pretectum, and ventral mesencephalon, and discrete
cell layers in the hippocampus, cerebellum, and olfactory bulb. Patterns of
hybridization included neuronal, glial, and ependymal cells, although white
matter was uniformly negative, as were most cerebrovascular endothelial
cells. Expression of TR mRNA by astroglia and dopaminergic neurons was
confirmed by colocalization with immunoreactivity for glial fibrillary
acidic protein (GFAP) in hippocampus and tyrosine hydroxylase in the
substantia nigra. Comparison between TR and prothrombin (thrombin's
precursor) cRNA hybridization demonstrated distinct but overlapping brain
distributions of these transcripts, most clearly evident in postnatally
developing, laminated structures. These results suggest widespread
utilization of, and multiple physiologic, and possibly pathophysiologic,
functions for, the thrombin/TR cell signaling system in the CNS.
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