GROWTH HORMONE-RELEASING HEXAPEPTIDE ELEVATES INTRACELLULAR CALCIUM IN RAT SOMATOTROPES BY 2 MECHANISMS

The actions of GH-releasing hexapeptide (His-D-Trp-Ala-Trp-D-Phe-Lys-N H2 or GHRP-6) on single rat somatotropes were studied using whole cell patch clamp electrophysiology and indo-1 Ca2+ photometry. GHRP-6 elev ated intracellular free Ca2+ ([Ca2+](i)) in two phases: a rapid transi ent phase, followed by a persistent phase. Based on its insensitivity to treatments that block Ca2+ entry [removal of external Ca2+, additio n of the dihydropyridine Ca2+ channel blocker nitrendipine (1 mu M), a nd the hyperpolarizing action of zero external Na+ or 100 nM somatosta tin], the transient elevation is the result of release of Ca2+ from in tracellular stores. The half-maximal concentration for the peak [Ca2+] (i) rise during Ca2+ release was 49 nM GHRP-6, Prior treatment of cell s with caffeine (10 mM) or ryanodine (50 mu M) abolished or partially occluded GHRP-6-induced Ca2+ release. Simultaneous measurement of [Ca2 +](i) and membrane current or potential revealed that the transient re lease of Ca2+ by GHRP-6 activates a voltage-independent Ca2+-activated K+ conductance, which transiently hyperpolarizes the somatotrope. The GHRP-6-induced persistent [Ca2+](i) elevation is abolished by removal of external Ca2+ or external Na+ or the addition of 1 mu M nitrendipi ne or 100 nM somatostatin, consistent with Ca2+ entry through voltage- dependent Ca2+ channels. In nondialyzed cells (perforated patch record ing), we have identified a long-lasting GHRP-6-induced depolarization which may be responsible for the persistent[Ca2+](i) elevation.