title: Src modulates contractile vascular smooth muscle function via regulation of focal adhesions.
abstract: Src is a known regulator of focal adhesion turnover in migrating cells; but, in contrast, Src is generally assumed to play little role in differentiated, contractile vascular smooth muscle (dVSM). The goal of the present study was to determine if Src-family kinases regulate focal adhesion proteins and how this might affect contractility of non-proliferative vascular smooth muscle. We demonstrate here, through the use of phosphotyrosine screening, deconvolution microscopy imaging, and differential centrifugation, that the activity of Src family kinases in aorta is regulated by the alpha agonist and vasoconstrictor phenylephrine, and leads to focal adhesion protein phosphorylation and remodeling in dVSM. Furthermore, Src inhibition via morpholino knockdown of Src or by the small molecule inhibitor PP2 prevents phenylephrine-induced adhesion protein phosphorylation, markedly slows the tissue's ability to contract, and decreases steady state contractile force amplitude. Significant vasoconstrictor-induced and Src-dependent phosphorylation of Cas pY-165, FAK pY-925, paxillin pY-118, and Erk1/2 were observed. However, increases in FAK 397 phosphorylation were not seen, demonstrating differences between cells in tissue versus migrating, proliferating cells. We show here that Src, in a cause and effect manner, regulates focal adhesion protein function and, consequently, modulates contractility during the action of a vasoconstrictor. These data point to the possibility that vascular focal adhesion proteins may be useful drug discovery targets for novel therapeutic approaches to cardiovascular disease.
authors.affiliation: Department of Health Sciences, Boston University, Boston, MA 02215, USA.
authors.forename: Jianghong
authors.initials: J
authors.lastname: Min
authors.forename: Maya
authors.initials: M
authors.lastname: Reznichenko
authors.forename: Ransom H
authors.initials: RH
authors.lastname: Poythress
authors.forename: Cynthia M
authors.initials: CM
authors.lastname: Gallant
authors.forename: Susanne
authors.initials: S
authors.lastname: Vetterkind
authors.forename: Yunping
authors.initials: Y
authors.lastname: Li
authors.forename: Kathleen G
authors.initials: KG
authors.lastname: Morgan
mesh_terms: D000818:Animals; D001011:Aorta / Q000502:physiology; D005289:Ferrets; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism*; D022001:Focal Adhesions / Q000187:drug effects / Q000502:physiology*; D055785:Gene Knockdown Techniques; D006801:Humans; D009119:Muscle Contraction / Q000187:drug effects / Q000502:physiology*; D009131:Muscle, Smooth, Vascular / Q000502:physiology*; D009924:Organ Culture Techniques; D010656:Phenylephrine / Q000494:pharmacology; D010766:Phosphorylation / Q000187:drug effects; D019000:Phosphotyrosine / Q000378:metabolism; D011743:Pyrimidines / Q000494:pharmacology; D015398:Signal Transduction; D019061:src-Family Kinases / Q000502:physiology
chemical_list: C412373:AG 1879; D011743:Pyrimidines; D010656:Phenylephrine; D019000:Phosphotyrosine; D051416:Focal Adhesion Protein-Tyrosine Kinases; D019061:src-Family Kinases