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RESEARCH COMMUNICATION |
a Franz-Volhard Clinic at the Max-Delbrück Center for Molecular Medicine, Humboldt University of Berlin, Germany
b Max Delbrück Center for Molecular Medicine, Berlin-Buch, Germany
Despite intensive interest in understanding the differentiation of vascular smooth muscle cells (VSMC), no information is available about differential regulation of ion channels in these cells. Since expression of the L-type Ca2+ channel can be influenced by differentiation in other cell types, we tested the hypothesis that the L-type (C class) channel is a specific differentiation marker of VSMC and that expression of these channels depends on the state of cell differentiation. We used rat aortic (A7r5) VSMC, which express functional L-type Ca2+ channels, and induced dedifferentiation by cell culture in different media. Treatment with retinoic acid was used to redifferentiate the VSMC. We characterized the differentiated state of the cells by using immunohistochemistry and Western blot analysis for smooth muscle (SM) 
-actin and SM-myosin heavy chain (MHC). The number of functional Ca2+ channels was significantly decreased in dedifferentiated VSMC and increased upon differentiation with retinoic acid. Ca2+ channel function was assessed by whole-cell voltage clamp techniques. Using Western blot and dihydropyridine binding analysis, we found that the expression of the Ca2+ channel 1 subunit, and to a lesser extent the ß2 subunit, was directly correlated with the expression of SM -actin and SM-MHC. We conclude that expression of L-type Ca2+ channel 1 subunits, and thus a functional Ca2+ channel, is highly coordinated with expression of the SM-specific proteins required for specialized smooth muscle cell functions. Furthermore, our results demonstrate that the L-type Ca2+ channel is a novel marker for differentiation of VSMC. The data suggest that regulation of ion channel expression during differentiation may have physiological importance for normal smooth muscle function and may influence VSMC behavior under pathophysiological conditions.—Gollasch, M., Haase, H., Ried, C., Lindschau, C., Morano, I., Luft, F. C., Haller, H. L-type calcium channel expression depends on the differentiated state of vascular smooth muscle cells. FASEB J. 12, 593–601 (1998)
Key Words: voltage-dependent Ca2+ channels • dihydropyridines • arterial smooth muscle • differentiation • retinoic acid • A7r5 cells • atherosclerosis
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