Dependence of Ca2+ channel currents on endogenous and exogenous sources of ATP in portal vein smooth muscle.

Abstract

Metabolic pathways in vascular smooth muscles (VSM) appear to be functionally compartmentalized such that glycolysis fuels membrane-related processes, whereas oxidative processes fuel actin-myosin interaction. Because ATP influences Ca2+ channel activity, we examined the effects of ATP and metabolic substrates on Ca2+ channel activity with patch-clamp techniques in VSM cells isolated from rat portal vein. The peak magnitude of the Ca2+ channel currents was found to depend on the ATP concentration in the patch pipette. Cells perfused with 1, 3, and 5 mMATP had mean peak currents of 4.7 +/- 0.6, 12.2 +/- 1.9, and 17.6 +/- 2.0 pA/pF, respectively, and all currents showed substantial rundown. In separate experiments performed in the absence of intracellular ATP, provision of glycolytic but not oxidative substrates was able to maintain Ca2+ channel currents at levels comparable with those seen in the presence of 1 mM ATP. In the presence of 5 mM ATP, provision of glycolytic substrates resulted in a high peak current amplitude that was also very stable. Finally, metabolic inhibition with cyanide and iodoacetate caused a significant increase in the rate of current rundown, even in the presence of 5 mM ATP. These findings indicate that Ca2+ channel current is strongly dependent on ATP and that the source of ATP can also be an important factor. Compared with exogenous provision of ATP, endogenous metabolism preferentially maintained Ca2+ channel currents, consistent with the hypothesis of a functionally separate subsarcolemmal compartment. This provides an effective pathway for linking E-C coupling and vascular contractility to the metabolic state of the vascular cell.