Evidence for localized calcium mobilization and influx in single rat gonadotropes.

Abstract

Dynamic video-imaging microscopy was used to investigate the spatial and temporal nature of Ca2+ mobilization and Ca2+ influx in acutely dissociated, fura-2-loaded, rat gonadotropes. Addition of luteinizing hormone-releasing hormone (LHRH) to an isolated gonadotrope stimulated a wave of Ca2+ originating from a specific locus of the cell. This probably reflects Ca2+ mobilization from an intracellular store, since this response was unaffected by the removal of extracellular Ca2+. Application of the dihydropyridine-sensitive Ca2+ channel agonist Bay K 8644 (Bay K) stimulated a rise in cytosolic free Ca2+ concentration in the rat gonadotrope. This response was blocked by the removal of extracellular Ca2+ and probably reflects the influx of Ca2+ across the cell membrane. High speed (30 frames.s-1) imaging of the Bay K-induced Ca2+ influx revealed a wave of Ca2+ originating from a localized part of the cell membrane, which, in general, was spatially distinct from the LHRH-induced Ca2+ wave produced in the same cell. This suggests that Ca2+ channels in the cell membrane may be clustered in a specific area of the cell membrane. The velocity of the LHRH-induced Ca2+ mobilization wave was faster (mean = 79 +/- 5 microns.s-1, n = 9) than the Bay K-induced Ca2+ influx wave (39 +/- 7 microns.s-1, n = 9) (p less than or equal to 0.01, Wilcoxon signed rank test) measured in the same cells. Thus, both Ca2+ mobilization from intracellular stores and Ca2+ influx through the cell membrane appear to be spatially localized in the rat gonadotrope. These findings may have important implications in the intracellular regulation of Ca(2+)-dependent cell functions such as hormone biosynthesis and secretion.