VOLTAGE-INDEPENDENT CALCIUM CHANNELS MEDIATE SLOW OSCILLATIONS OF CYTOSOLIC CALCIUM THAT ARE GLUCOSE-DEPENDENT IN PANCREATIC BETA-CELLS

Pancreatic beta-cells and HIT-T15 cells exhibit oscillations of cytosolic calcium ([Ca2+](i)) that are dependent on glucose metabolism and appear to trigger pulsatile insulin secretion. Significantly, differences in the pattern of this [Ca2+](i) oscillatory activity may have important implications for our understanding of how glucose homeostasis is achieved during the feeding and fasting states. When single beta-cells are exposed to a stepwise increase in glucose concentration that mimics the transition from fasting to feeding states, fast irregular oscillations of [Ca2+](i) are observed. Alternatively, when single beta-cells are equilibrated in a steady state concentration of glucose that mimics the fasting state, slow periodic oscillations of [Ca2+](i) are noted. Here we report a fundamental difference in the mechanism by which glucose induces these two types of [Ca2+](i) oscillatory activity. In agreement with previous studies, we substantiate a role for L-type voltage-dependent Ca2+ channels as mediators of the fast oscillations of [Ca2+](i) observed after a stepwise increase in glucose concentration. In marked contrast, we report that voltage-independent calcium channels (VICCs) mediate slow oscillations of [Ca2+](i) that occur when beta-cells are equilibrated in steady state concentrations of glucose. Slow [Ca2+](i) oscillations are mediated by VICCs which are pharmacologically and biophysically distinguishable from voltage-dependent Ca2+ channels that mediate fast oscillations. Specifically, slow [Ca2+](i) oscillations are blocked by extracellular La3+, but not by nifedipine, and are independent of changes in membrane potential. Measurements of membrane conductance also indicate an important role for VICCs, as demonstrated by a steady state inward Ca2+ current that is blocked by La3+. The steady state Ca2+ current appears to generate slow [Ca2+](i) oscillations by triggering Ca2+-induced Ca2+ release from intracellular Ca2+ stores, a process that is mimicked by extracellular application of caffeine, a sensitizer of the ryanodine receptor/Ca2+ release channel. Depletion of intracellular Ca2+ stores with thapsigargin stimulated Mn2+ influx, suggesting the presence of Ca2+-release-activated Ca2+ channels. Taken together, these observations are consistent with a role for VICCs (possibly G-type channels) and/or Ca2+-release-activated Ca2+ channels as mediators of slow [Ca2+](i) oscillations in beta-cells. We propose that slow oscillations of [Ca2+](i) probably serve as important initiators of insulin secretion under conditions in which tight control of glucose homeostasis is necessary, as is the case during the fasting normoglycemic state.