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In smooth muscles, inhibition of Na+,K+,2Cl- cotransport (NKCC) by bumetanide decreased intracellular Cl- content ([Cl-]i) and suppressed the contractions triggered by diverse stimuli. This study examines whether or not bicarbonate, a regulator of several Cl- transporters, affects the impact of NKCC in excitation-contraction coupling. Addition of 25 mM NaHCO3 attenuated the inhibitory action of bumetanide on mesenteric artery contractions evoked by 30 mM KCl and phenylephrine (PE) by 5 and 3-fold, respectively. In cultured vascular smooth muscle cells, NaHCO3 almost completely abolished inhibitory actions of bumetanide on transient depolarization and [Ca2+]i elevation triggered by PE. In bicarbonate-free medium, bumetanide decreased [Cl-]i by ∼15%; this effect was almost totally abrogated by NaHCO3. The addition of NaHCO3 resulted in 2-fold inhibition of NKCC activity and 3-fold attenuation of [Cl-]i. These data strongly suggest that extracellular HCO3- diminishes the NKCC-sensitive component of excitation-contraction coupling via suppression of this carrier. Copyright © 2009 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Previously, we reported that hyposmotic swelling evoked transient vascular smooth muscle cell (SMC) contraction that was completely abolished by L-type Ca2+ channel blockers. In contrast, sustained contraction revealed in hyper- and isoosmotically-shrunken SMCs was insensitive to L-type channel blockers and was diminished in Ca2+-free medium by only 30-50%. Several research groups reported cell volume-dependent cytoskeleton network rearrangements. This study examines the role of cytoskeleton proteins in cell volume-dependent contraction of endothelium-denuded vascular smooth muscle rings (VSMR) from the rat thoracic aorta. Hyperosmotic shrinkage and hyposmotic swelling were triggered by modulation of medium osmolality; isosmotic shrinkage was induced by VSMR transfer from hypo- to isosmotic medium. The relative content of globular (G) and fibrillar (F) actin was estimated by fluorescence microscopy. Hyperosmotic shrinkage and hyposmotic swelling led to elevation of the F-actin/G-actin ratio by 2.5- and 1.8-fold respectively. Contraction of shrunken and swollen VSMR was insensitive to modulators of microtubules such as vinblastine, colchicine and docetaxel. Microfilament disassembly by cytochalasin B resulted in dramatic attenuation of the maximal amplitude of contraction of hyperosmotically-shrunken and hyposmotically-swollen VSMR, and almost completely abolished the contraction triggered by isosmotic shrinkage. These data suggest that both L-type Ca2+ channel-mediated contraction of swollen vascular SMC and Ca2+o-insensitive contractions of shrunken cells are triggered by reorganization of the microfilament network caused by elevation of the F-actin/G-actin ratio. Copyright © 2008 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]