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Am J Physiol Heart Circ Physiol (September 23, 2004). doi:10.1152/ajpheart.00608.2004
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Submitted on June 18, 2004
Accepted on September 20, 2004

IMAGING REMODELING OF THE ACTIN CYTOSKELETON IN VASCULAR SMOOTH MUSCLE CELLS FOLLOWING MECHANOSENSITIVE ARTERIOLAR CONSTRICTION

Nicholas A. Flavahan*, Simon R. Bailey, William A. Flavahan, Srabani Mitra, and Sheila Flavahan

* To whom correspondence should be addressed. E-mail: flavahan-1{at}medctr.osu.edu.

Experiments were performed to determine whether remodeling of the actin cytoskeleton contributes to arteriolar constriction. Mouse tail arterioles were mounted on cannulae in a myograph and superfused with buffer solution. The {alpha}1-adrenergic agonist phenylephrine (0.1 to 1 µmol/L) caused constriction that was unaffected by cytochalasin D (300 nmol/L) or latrunculin A (100 nmol/L), inhibitors of actin polymerization. In contrast, each compound abolished the mechanosensitive constriction (myogenic response) evoked by elevation in transmural pressure (PTM; 10 to 60 or 90 mmHg). Arterioles were fixed, permeabilized and stained with Alexa-568 phalloidin and Alexa-488 DNAse I to visualize F-actin and G-actin, respectively, using a Zeiss 510 laser scanning microscope. Elevation in PTM, but not phenylephrine (1 µmol/L), significantly increased the intensity of F-actin and significantly decreased the intensity of G-actin staining in arteriolar vascular smooth muscle cells (VSMs). The increase in F-actin staining caused by elevation in PTM was inhibited by cytochalasin D. In VSMs at 10 mmHg, prominent F-actin staining was restricted to the cell periphery, whereas after elevation in PTM, transcytoplasmic F-actin fibers were localized through the cell interior, running parallel to the long axis of the cells. Phenylephrine (1 µmol/L) did not alter the architecture of the actin cytoskeleton. In contrast to VSMs, the actin cytoskeleton of endothelial or adventitial cells was not altered by elevation in PTM. Therefore, the actin cytoskeleton of VSMs undergoes dramatic alteration following elevation in PTM of arterioles and plays a selective and essential role in mechanosensitive, myogenic constriction.




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