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1 Department of Molecular and Cellular Physiology, College of Medicine, University of Cincinnati, Cincinnati, Ohio 45267-0576; and 2 First Department of Physiology, School of Medicine, University of Ryukyus, Nishihara, Okinawa 903-0215, Japan
To clarify
interactions between the cytoskeleton and activity of L-type
Ca2+ (CaL) channels in vascular smooth muscle
(VSM) cells, we investigated the effect of disruption of actin
filaments and microtubules on the L-type Ca2+ current
[IBa(L)] of cultured VSM cells (A7r5 cell
line) using whole cell voltage clamp. The cells were exposed to each
disrupter for 1 h and then examined electrophysiologically and
morphologically. Results of immunostaining using anti-
-actin and
anti--tubulin antibodies showed that colchicine disrupted both actin
filaments and microtubules, cytochalasin D disrupted only actin
filaments, and nocodazole disrupted only microtubules.
IBa(L) was greatly reduced in cells that were
exposed to colchicine or cytochalasin D but not to nocodazole.
Colchicine even inhibited IBa(L) by about 40%
when the actin filaments were stabilized by phalloidin or when the
cells were treated with phalloidin plus taxol to stabilize both
cytoskeletal components. These results suggest that colchicine must
also cause some inhibition of IBa(L) due to
another unknown mechanism, e.g., a direct block of CaL
channels. In summary, actin filament disruption of VSM cells inhibits
CaL channel activity, whereas disrupting the microtubules
does not.
cytoskeleton; microtubules; colchicine; cytochalasin D; nocodazole
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