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Effect of calcium and cAMP on Go alpha expression in neonatal rat cardiac myocytes
1Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania 17822.
Abstract:
Culturing neonatal rat cardiac myocytes in 50 mM KCl inhibits the accumulation of Go alpha that occurs when myocytes are placed in culture. The mechanism by which high extracellular K+ inhibits Go alpha accumulation in myocytes was investigated by measurement of the concentration of intracellular Ca2+ ([Ca2+]) and adenosine 3',5'-cyclic monophosphate concentration ([cAMP]) of control and K(+)-depolarized myocytes. Although intracellular [Ca2+] in K(+)-depolarized myocytes was twofold higher than basal intracellular [Ca2+] in control cells, the mean intracellular [Ca2+] in contracting control myocytes was comparable to that of K(+)-depolarized myocytes. Furthermore, myocytes cultured in low Ca2+ plus high K+ exhibited an inhibition of Go alpha accumulation, even though intracellular [Ca2+] was 10-fold lower than that of cells cultured in normal Ca2+ plus high K+. In addition, intracellular [cAMP] of K(+)-depolarized myocytes was comparable to that of control cells. Moreover, dibutyryl cAMP inhibited Go alpha accumulation in myocytes to the same extent as high K+, even though intracellular [cAMP] differed 10-fold. Thus neither intracellular Ca2+ nor cAMP appear to mediate the inhibitory effect of high K+ on Go alpha accumulation. However, cAMP has an inhibitory effect on Go alpha expression that is independent of K+.