Glucose inhibits GABA release by pancreatic [beta]-cells through an increase in GABA shunt activity

GABA is the major inhibitory neurotransmitter in the nervous system. It is also released by the insulin-producing [beta]-cells, providing them with a potential paracrine regulator. Because glucose was found to inhibit GABA release, we investigated whether extracellular GABA can serve as a marker for glucose-induced mitochondrial activity and thus for the functional state of [beta]-cells. GABA release by rat and human [beta]-cells was shown to reflect net GABA production, varying with the functional state of the cells. Net GABA production is the result of GABA formation through glutamate decarboxylase (GAD) and GABA catabolism involving a GABA-transferase (GABA-T)-mediated shunt to the TCA cycle. GABA-T exhibits [K.sub.m] values for GABA (1.25 mM) and for [alpha]-ketoglutarate ([alpha]-KG; 0.49 mM) that are, respectively, similar to and lower than those in brain. The GABA-T inhibitor [gamma]-vinyl GABA was used to assess the relative contribution of GABA formation and catabolism to net production and release. The nutrient status of the [beta]-cells was found to regulate both processes. Glutamine dose-dependently increased GAD-mediated formation of GABA, whereas glucose metabolism shunts part of this GABA to mitochondrial catabolism, involving [alpha]-KG-induced activation of GABA-T. In absence of extracellular glutamine, glucose also contributed to GABA formation through aminotransferase generation of glutamate from [alpha]-KG; this stimulatory effect increased GABA release only when GABA-T activity was suppressed. We conclude that GABA release from [beta]-cells is regulated by glutamine and glucose. Glucose inhibits glutaminedriven GABA formation and release through increasing GABA-T shunt activity by its cellular metabolism. Our data indicate that GABA release by [beta]-cells can be used to monitor their metabolic responsiveness to glucose irrespective of their insulin-secretory activity. glutamine; [gamma]-aminobutyric acid; [gamma]-aminobutyric acid transferase