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L-type channel antagonists are of therapeutic benefit in the treatment of hyperlipidaemia and insulin resistance. Our aim was to identify L-type voltage-gated Ca 2+  channels in white fat adipocytes, and determine if they affect intracellular Ca 2+ , lipolysis and lipogenesis. We used a multidisciplinary approach of molecular biology, confocal microscopy, Ca 2+  imaging and metabolic assays to explore this problem using adipocytes isolated from adult rat epididymal fat pads. CaV1.2, CaV1.3 and CaV1.1 alpha 1 , beta and alpha 2 delta subunits were detected at the gene expression level. The CaV1.2 and CaV1.3 alpha1 subunits were identified in the plasma membrane at the protein level. Confocal microscopy with fluorescent antibodies labelled CaV1.2 in the plasma membrane. Ca 2+  imaging revealed that the intracellular Ca 2+  concentration, [Ca 2  + ] i  was reversibly decreased by removal of extracellular Ca 2+ , an effect mimicked by verapamil, nifedipine and Co 2+ , all blockers of L-type channels, whereas the Ca 2+  channel agonist BAY-K8644 increased [Ca 2+ ] i . The finding that the magnitude of these effects correlated with basal [Ca 2+ ] i  suggests that adipocyte [Ca 2+ ] i  is controlled by L-type Ca 2+  channels that are constitutively active at the adipocyte depolarized membrane potential. Pharmacological manipulation of L-type channel activity modulated both basal and catecholamine-stimulated lipolysis but not insulin-induced glucose uptake or lipogenesis. We conclude that white adipocytes have constitutively active L-type Ca 2+  channels which explains their sensitivity of lipolysis to Ca 2+  channel modulators. Our data suggest CaV1.2 as a potential novel therapeutic target in the treatment of obesity.

CaV1.2 and CaV1.3 voltage-gated L-type Ca2+ channels in rat white fat adipocytes