Origin of ATP for Ca 2+ -induced contraction in the guinea-pig femoral artery

Abstract

Previously, we have described differences between the rat proximal colon and femoral artery with respect to the role of ATP newly synthesized by creatine kinase. In the present study the role of newly synthesized ATP was studied in the guinea-pig femoral artery to examine species differences. In the alpha-toxin-permeabilized preparation of the guinea-pig femoral artery, the rapid Ca(2+)-induced contraction was suppressed when creatine kinase activity was inhibited. The contraction was restored completely by treatment with NaN(3), an inhibitor of ecto-ATPase, the enzyme that breaks down exogenous ATP. Thus, ATP newly synthesized by creatine kinase may have no role in contraction of the guinea-pig femoral artery. This is in marked contrast to the rat femoral artery, in which Ca(2+)-induced contractions are almost completely inhibited by inhibition of creatine kinase activity but only partly restored by NaN(3). To characterize the difference between the guinea-pig and rat tissue, the origin of ATP required for contraction was determined in intact preparations. Monoiodoacetic acid, an inhibitor of glycolysis, inhibited the high K(+)-induced contraction in the guinea-pig femoral artery more potently than in the rat tissue. In contrast, an inhibitor of mitochondrial respiration, carbonylcyanide p-(trifluoromethoxy)phenylhydrazone (FCCP), inhibited contraction in femoral arteries from rats, but not from guinea-pigs. These results suggest that contraction in the rat femoral artery is dependent largely on oxidative phosphorylation, while contraction in the guinea-pig tissue is dependent only on glycolysis. Because oxidative phosphorylation generates ATP and phosphocreatine, while glycolysis generates only ATP, the strong dependence of the contraction of the rat femoral artery on the oxidative phosphorylation is consistent with its dependence on ATP newly synthesized by creatine kinase from ADP and phosphocreatine, as previously shown. Thus, it is proposed that ATP, newly synthesized by creatine kinase, in addition to ATP generated by oxidative phosphorylation, is utilized for contraction in the rat femoral artery, while glycolysis produces sufficient ATP for contraction in the guinea-pig femoral artery.