Sex‐Specific Differences in Endothelial Function Are Driven by Divergent Mitochondrial Ca(2+) Handling

BACKGROUND: Sex‐specific differences in vasodilation are mediated in part by differences in cytosolic Ca(2+) handling, but how variations in mitochondrial Ca(2+) contributes to this effect remains unknown. Here, we investigated the extent to which mitochondrial Ca(2+) entry via the MCU (mitochondrial Ca(2+) uniporter) drives sex differences in vasoreactivity in resistance arteries. METHODS AND RESULTS: Enhanced vasodilation of mesenteric resistance arteries to acetylcholine (ACh) was reduced to larger extent in female compared with male mice in 2 genetic models of endothelial MCU ablation. Ex vivo Ca(2+) imaging of mesenteric arteries with Fura‐2AM confirmed higher cytosolic Ca(2+) transients triggered by ACh in arteries from female mice versus male mice. MCU inhibition both strongly reduced cytosolic Ca(2+) transients and blocked mitochondrial Ca(2+) entry. In cultured human aortic endothelial cells, treatment with physiological concentrations of estradiol enhanced cytosolic Ca(2+) transients, Ca(2+) buffering capacity, and mitochondrial Ca(2+) entry in response to ATP or repeat Ca(2+) boluses. Further experiments to establish the mechanisms underlying these effects did not reveal significant differences in the expression of MCU subunits, at either the mRNA or protein level. However, estradiol treatment was associated with an increase in mitochondrial mass, mitochondrial fusion, and the mitochondrial membrane potential and reduced mitochondrial superoxide production. CONCLUSIONS: Our data confirm that mitochondrial function in endothelial cells differs by sex, with female mice having enhanced Ca(2+) uptake capacity, and that these differences are attributable to the presence of more mitochondria and a higher mitochondrial membrane potential in female mice rather than differences in composition of the MCU complex.