Cerebrovascular connexin expression: Effects of traumatic brain injury

Marcela A. Avila, Stacy L. Sell, Bridget E. Hawkins, Helen L. Hellmich, Debbie R. Boone, Jeanna M. Crookshanks, Donald S. Prough, Douglas S. Dewitt

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

Traumatic brain injury (TBI) results in dysfunction of the cerebrovasculature. Gap junctions coordinate vasomotor responses and evidence suggests that they are involved in cerebrovascular dysfunction after TBI. Gap junctions are comprised of connexin proteins (Cxs), of which Cx37, Cx40, Cx43, and Cx45 are expressed in vascular tissue. This study tests the hypothesis that TBI alters Cx mRNA and protein expression in cerebral vascular smooth muscle and endothelial cells. Anesthetized (1.5% isoflurane) male Sprague-Dawley rats received sham or fluid-percussion TBI. Two, 6, and 24h after, cerebral arteries were harvested, fresh-frozen for RNA isolation, or homogenized for Western blot analysis. Cerebral vascular endothelial and smooth muscle cells were selected from frozen sections using laser capture microdissection. RNA was quantified by ribonuclease protection assay. The mRNA for all four Cx genes showed greater expression in the smooth muscle layer compared to the endothelial layer. Smooth muscle Cx43 mRNA expression was reduced 2h and endothelial Cx45 mRNA expression was reduced 24h after injury. Western blot analysis revealed that Cx40 protein expression increased, while Cx45 protein expression decreased 24h after injury. These studies revealed significant changes in the mRNA and protein expression of specific vascular Cxs after TBI. This is the first demonstration of cell type-related differential expression of Cx mRNA in cerebral arteries, and is a first step in evaluating the effects of TBI on gap junction communication in the cerebrovasculature.

Original languageEnglish (US)
Pages (from-to)1803-1811
Number of pages9
JournalJournal of neurotrauma
Volume28
Issue number9
DOIs
StatePublished - Sep 1 2011

Keywords

  • connexin
  • endothelial cells
  • gap junction
  • laser capture microdissection
  • traumatic brain injury

ASJC Scopus subject areas

  • Clinical Neurology

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