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J Appl Physiol 93: 1907-1917, 2002. First published August 30, 2002; doi:10.1152/japplphysiol.00988.2001
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Vol. 93, Issue 6, 1907-1917, December 2002

Smooth muscle cells contract in response to fluid flow via a Ca2+-independent signaling mechanism

Mete Civelek, Kristy Ainslie, Jeff S. Garanich, and John M. Tarbell

Biomolecular Transport Dynamics Laboratory, Departments of Chemical Engineering and Bioengineering, The Pennsylvania State University, University Park, Pennsylvania 16802

Smooth muscle cells (SMC) are exposed to fluid shear stress because of transmural (interstitial) flow across the arterial wall. This shear stress may play a role in the myogenic response and flow-mediated vasomotion. We, therefore, examined the effects of fluid flow on contraction of rat aortic SMC. SMC that had been serum-starved to induce a contractile phenotype were plated on quartz slides and exposed to controlled shear stress levels in a flow chamber. The area of the cells was quantified, and reduction in the cell area was reported as contraction. At 25 dyn/cm2, significant area reduction was apparent 3 min after the onset of flow and exceeded 30% at 30 min. At 1 dyn/cm2, significant contraction was not observed at 30 min. The threshold for significant shear-induced contraction appeared to be 11 dyn/cm2. The signal transduction mechanism was studied at 25 dyn/cm2. Intracellular calcium was imaged by using the calcium-sensitive fluorescent dye fura 2-AM. There was no detectable change in intracellular calcium during 10 min of exposure to shear stress, even though the cells displayed a significant calcium response to thapsigargin, calcium ionophore, and KCl. Further studies using pathway inhibitors provided evidence that the most important signal transduction pathway mediating calcium-independent contraction in response to fluid flow is the Rho-kinase pathway, although there was a suggestion that protein kinase C plays a secondary role.

shear stress; vascular smooth muscle; myogenic response; calcium-independent contraction


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