Vasoconstrictor-induced endocytic recycling regulates focal adhesion protein localization and function in vascular smooth muscle
| Autor: | Cynthia Gallant, Susanne Vetterkind, Kathleen G. Morgan, Ransom H. Poythress |
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| Rok vydání: | 2013 |
| Předmět: |
Male
Vascular smooth muscle Physiology Myocytes Smooth Muscle Endocytic recycling Muscle Smooth Vascular Zyxin Focal adhesion chemistry.chemical_compound Phenylephrine Animals Vasoconstrictor Agents Paxillin Cells Cultured Cytochalasin D biology Microfilament Proteins Ferrets Cell migration Cell Biology Articles Vinculin Phosphoproteins Endocytosis Cell biology Protein Transport chemistry Gene Expression Regulation Focal Adhesion Protein-Tyrosine Kinases biology.protein Cell Adhesion Molecules |
| Zdroj: | American journal of physiology. Cell physiology. 305(2) |
| ISSN: | 1522-1563 |
| Popis: | Turnover of focal adhesions (FAs) is known to be critical for cell migration and adhesion of proliferative vascular smooth muscle (VSM) cells. However, it is often assumed that FAs in nonmigratory, differentiated VSM (dVSM) cells embedded in the wall of healthy blood vessels are stable structures. Recent work has demonstrated agonist-induced actin polymerization and Src-dependent FA phosphorylation in dVSM cells, suggesting that agonist-induced FA remodeling occurs. However, the mechanisms and extent of FA remodeling are largely unknown in dVSM. Here we show, for the first time, that a distinct subpopulation of dVSM FA proteins, but not the entire FA, remodels in response to the α-agonist phenylephrine. Vasodilator-stimulated phosphoprotein and zyxin displayed the largest redistributions, while β-integrin and FA kinase showed undetectable redistribution. Vinculin, metavinculin, Src, Crk-associated substrate, and paxillin displayed intermediate degrees of redistribution. Redistributions into membrane fractions were especially prominent, suggesting endosomal mechanisms. Deconvolution microscopy, quantitative colocalization analysis, and Duolink proximity ligation assays revealed that phenylephrine increases the association of FA proteins with early endosomal markers Rab5 and early endosomal antigen 1. Endosomal disruption with the small-molecule inhibitor primaquine inhibits agonist-induced redistribution of FA proteins, confirming endosomal recycling. FA recycling was also inhibited by cytochalasin D, latrunculin B, and colchicine, indicating that the redistribution is actin- and microtubule-dependent. Furthermore, inhibition of endosomal recycling causes a significant inhibition of the rate of development of agonist-induced dVSM contractions. Thus these studies are consistent with the concept that FAs in dVSM cells, embedded in the wall of the aorta, remodel during the action of a vasoconstrictor. |
| Databáze: | OpenAIRE |
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