The role of mitochondria in stimulus–secretion coupling of pancreatic β-cells was examined using methyl pyruvate (MP). MP stimulated insulin secretion in the absence of glucose, with maximal effect at 5mM. K+ (30mM) alone, or in combination with diazoxide (100µM), failed to enhance MP-induced secretion. Diazoxide (100µM) inhibited MP-induced insulin secretion. MP depolarized the β-cell in a concentration-dependent manner (5–20mM). The sustained depolarization induced by 20mM MP was not influenced by 100µM diazoxide, but the continuous spiking activity was suppressed by 500µM diazoxide. Pyruvate failed to initiate insulin release (5–20mM) or to depolarize the membrane potential. ATP production in isolated β-cell mitochondria was detected as accumulation of ATP in the medium during incubation in the presence of malate or glutamate in combination with pyruvate or MP. There was no difference in ATP production induced by pyruvate/malate or MP/malate in isolated β-cell mitochondria. ATP production by MP/glutamate was higher than that induced by pyruvate/glutamate, but it was much lower than that induced by α-ketoisocaproate/glutamate. Pyruvate (5mM) or MP (5mM) had no effect on the ATP/ADP ratio in whole islets, whereas glucose (20mM) significantly increased the whole islet ATP/ADP ratio. It is concluded that MP-induced β-cell membrane depolarization or insulin release does not relate directly to mitochondrial ATP production. Instead MP may exert a direct extramitochondrial effect, or it may stimulate β-cell mitochondria to produce coupling factors different from ATP to initiate insulin release.

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