cGMP-dependent protein kinase expression restores contractile function in cultured vascular smooth muscle cells

Colleen M. Brophy, David A. Woodrum, Jennifer Pollock, Mary Dickinson, Padmini Komalavilas, Trudy L. Cornwell, Thomas M. Lincoln

Research output: Contribution to journalArticle

32 Scopus citations

Abstract

Vascular diseases, such as atherosclerosis and restenosis following angioplasty or transplantation, are due to abnormal vascular smooth muscle growth and gene expression. The smooth muscle cells (SMC) in response to injury lose their contractile function, become highly proliferative and synthesize and secrete extracellular matrix proteins. Similar changes in the phenotypic properties of vascular SMC occur during in vitro culture. In this report, we examined whether restoration of the expression of the major receptor protein for nitric oxide (NO) signaling in smooth muscle, the guanosine 3′:5′ cyclic monophosphate (cGMP)-dependent protein kinase (PKG), reestablished contractile function to cultured rat aortic SMC. Contractile function was monitored using the silicone polymer wrinkle assay used previously to determine contractility in cultured mesangial cells. Non-contractile rat aortic smooth muscle cells transfected with the cDNA encoding the type I isoform of PKG, but not those transfected with empty vector, formed discreet wrinkles on the substratum in response to serum indicative of contraction. Treatment of the PKG-expressing SMC with sodium nitroprusside (SNP), an NO donor, and with cGMP analogs, or with the adenylyl cyclase activator, forskolin, and with adenosine 3′:5′ cyclic monophosphate (cAMP) analogs reduced wrinkling. The expression of a major PKG substrate protein involved in smooth muscle relaxation, heat shock-related protein-20 (HSP20), was also reestablished in PKG-expressing SMC. Treatment of the PKG-expressing SMC with nitroprus-side resulted in phosphorylation of HSP20. Collectively, these results indicate that PKG expression is important to establish contractility to SMC in culture.

Original languageEnglish (US)
Pages (from-to)95-103
Number of pages9
JournalJournal of Vascular Research
Volume39
Issue number2
DOIs
StatePublished - May 23 2002

Keywords

  • Heat shock proteins
  • Phenotype
  • Phosphorylation
  • Protein kinase
  • Relaxation
  • Smooth muscle
  • cGMP

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

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