TY - JOUR
T1 - Role of calcium and EPAC in norepinephrine-induced ghrelin secretion
AU - Mani, Bharath K.
AU - Chuang, Jen Chieh
AU - Kjalarsdottir, Lilja
AU - Sakata, Ichiro
AU - Walker, Angela K.
AU - Kuperman, Anna
AU - Osborne-Lawrence, Sherri
AU - Repa, Joyce J.
AU - Zigman, Jeffrey M.
PY - 2014/1
Y1 - 2014/1
N2 - Ghrelin is an orexigenic hormone secreted principally from a distinct population of gastric endocrine cells. Molecular mechanisms regulating ghrelin secretion are mostly unknown. Recently, norepinephrine (NE) was shown to enhance ghrelin release by binding to1-Adrenergic receptors on ghrelin cells. Here,weuseanimmortalized stomach-derived ghrelin cell line to further characterize the intracellular signaling pathways involved in NE-induced ghrelin secretion, with a focus on the roles of Ca2+ and cAMP. Several voltage-gated Ca2+ channel (VGCC) family members were found by quantitative PCR to be expressed by ghrelin cells. Nifedipine, a selective L-type VGCC blocker, suppressed both basal and NE-stimulated ghrelin secretion.NEinduced elevation of cytosolic Ca2+levels both in the presence and absence of extracellular Ca2+. Ca2+-sensing synaptotagmins Syt7 and Syt9 were also highly expressed in ghrelin cell lines, suggesting that they too help mediate ghrelin secretion. Raising cAMP with the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine also stimulated ghrelin secretion, although such a cAMP-mediated effect likely does not involve protein kinase A, given the absence of a modulatory response to a highly selective protein kinase A inhibitor. However, pharmacological inhibition of another target of cAMP, exchange protein-Activated by cAMP (EPAC), did attenuate both basal and NE-inducedghrelin secretion,whereasanEPACagonistenhancedbasal ghrelin secretion.Weconclude that constitutive ghrelin secretion is primarily regulated by Ca2+ influx through L-type VGCCs and that NE stimulates ghrelin secretion predominantly through release of intracellular Ca2+. Furthermore, cAMP and its downstream activation of EPAC are required for the normal ghrelin secretory response to NE.
AB - Ghrelin is an orexigenic hormone secreted principally from a distinct population of gastric endocrine cells. Molecular mechanisms regulating ghrelin secretion are mostly unknown. Recently, norepinephrine (NE) was shown to enhance ghrelin release by binding to1-Adrenergic receptors on ghrelin cells. Here,weuseanimmortalized stomach-derived ghrelin cell line to further characterize the intracellular signaling pathways involved in NE-induced ghrelin secretion, with a focus on the roles of Ca2+ and cAMP. Several voltage-gated Ca2+ channel (VGCC) family members were found by quantitative PCR to be expressed by ghrelin cells. Nifedipine, a selective L-type VGCC blocker, suppressed both basal and NE-stimulated ghrelin secretion.NEinduced elevation of cytosolic Ca2+levels both in the presence and absence of extracellular Ca2+. Ca2+-sensing synaptotagmins Syt7 and Syt9 were also highly expressed in ghrelin cell lines, suggesting that they too help mediate ghrelin secretion. Raising cAMP with the phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine also stimulated ghrelin secretion, although such a cAMP-mediated effect likely does not involve protein kinase A, given the absence of a modulatory response to a highly selective protein kinase A inhibitor. However, pharmacological inhibition of another target of cAMP, exchange protein-Activated by cAMP (EPAC), did attenuate both basal and NE-inducedghrelin secretion,whereasanEPACagonistenhancedbasal ghrelin secretion.Weconclude that constitutive ghrelin secretion is primarily regulated by Ca2+ influx through L-type VGCCs and that NE stimulates ghrelin secretion predominantly through release of intracellular Ca2+. Furthermore, cAMP and its downstream activation of EPAC are required for the normal ghrelin secretory response to NE.
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U2 - 10.1210/en.2013-1691
DO - 10.1210/en.2013-1691
M3 - Article
C2 - 24189139
AN - SCOPUS:84891424484
SN - 0013-7227
VL - 155
SP - 98
EP - 107
JO - Endocrinology
JF - Endocrinology
IS - 1
ER -