Ghrelin is an endogenous ligand for growth hormone secretagogue receptor (GHS-R) and is predominantly produced by the stomach and lower amounts in the hypothalamus and various peripheral tissues. Ghrelin is a potent stimulator of growth hormone (GH) secretion from the pituitary in vivo and in vitro. GH secretion from the pituitary also is controlled by two hypothalamic peptides: stimulatory GH-releasing hormone (GHRH) and inhibitory somatostatin-14 (SRIH). GH participates in its own rhythmic secretion through feedback action on GHRH and SRIH neurons. The mechanism of action of GHS is not established. The present study examined the signal transduction pathways of ghrelin in isolated porcine somatotropes. The ability of ghrelin to induce an increase in the intracellular Ca2+ concentration – [Ca2+]i – somatotropes was examined in dispersed porcine pituitary cells using a calcium imaging system. Somatotropes were functionally identified by application of human growth hormone releasing hormone (hGHRH). Ghrelin increased the [Ca2+]i in a dose-dependent manner in 98% of the cells that responded to. In the presence of (D-Lys3)-GHRP-6, a specific receptor antagonist of GHS-R, the increase in [Ca2+]i evoked by ghrelin was decreased. Pretreatment of cultures with somatostatin or neuropeptide Y reduced the ghrelin-induced increase of [Ca2+]i. The stimulatory effect of ghrelin on somatotropes was greatly attenuated in lowcalcium saline and blocked by nifedipine, an L-type calcium channel blocker, suggesting involvement of calcium channels. In a zero Na+ solution, the stimulatory effect of ghrelin on somatotropes was decreased, suggesting that besides calcium channels, sodium channels are also involved in ghrelin-induced calcium transients. Either SQ-22536, an adenylyl cyclase inhibitor, or U73122, a phospholipase C inhibitor, decreased the stimulatory effects of ghrelin on [Ca2+]i transiently, indicating the involvement of adenylyl cyclase-cyclic adenosine monophosphate and phospholipase C inositol 1,4,5-trisphosphate pathways. The non-peptidyl GHS, L-692,585 (L-585), induced changes in [Ca2+]i similar to those observed with ghrelin. Application of L-585 after ghrelin did not have additive effects on [Ca2+]i. Preapplication of L-585 blocked the stimulatory effect of ghrelin on somatotropes. Our results suggest that the actions of ghrelin and synthetic GHS closely parallel each other, in a manner that is consistent with an increase of hormone secretion. An understanding of the molecular mechanisms by which ghrelin and GHS modulate GH secretion is of particular interest in the regulation of GH for muscle accretion and somatic growth.