Ghrelin reduces voltage-gated potassium currents in GH3 cells via cyclic GMP pathways.
Ghrelin is an endogenous growth hormone secretagogue (GHS) causing release of GH from pituitary somatotropes through the GHS receptor. Secretion of GH is linked directly to intracellular free Ca(2+) concentration ([Ca(2+)]i), which is determined by Ca(2+) influx and release from intracellular Ca(2+) storage sites. Ca(2+) influx is via voltage-gated Ca(2+) channels, which are activated by cell depolarization. Membrane potential is mainly determined by transmembrane K(+) channels. The present study investigates the in vitroeffect of ghrelin on membrane voltage-gated K(+) channels in the GH3 rat somatotrope cell line. Nystatin-perforated patch clamp recording was used to record K(+) currents under voltage-clamp conditions. In the presence of Co(2+) (1 mM, Ca(2+) channel blocker) and tetrodotoxin (1 microM, Na(+) channel blocker) in the bath solution, two types of voltage-gated K(+) currents were characterized on the basis of their biophysical kinetics and pharmacological properties. We observed that transient K(+) current (IA) represented a significant proportion of total K(+) currents in some cells, whereas delayed rectifier K(+) current (IK) existed in all cells. The application of ghrelin (10 nM) reversibly and significantly decreased the amplitude of both IA and IK currents to 48% and 64% of control, respectively. Application of apamin (1 microM, SK channel blocker) or charybdotoxin (1 microM, BK channel blocker) did not alter the K(+) current or the response to ghrelin. The ghrelin-induced reduction in K(+) currents was not affected by PKC and PKA inhibitors. KT5823, a specific PKG inhibitor, totally abolished the K+ current response to ghrelin. These results suggest that ghrelin-induced reduction of voltage-gated K(+) currents in GH3 cells is mediated through a PKG-dependent pathway. A decrease in voltage-gated K(+) currents may increase the frequency, duration, and amplitude of action potentials and contribute to GH secretion from somatotropes.
Han
Xue Feng
XF
Department of Physiology, Fourth Military Medical University, Shannxi, China.
Zhu
Yun Long
YL
Hernandez
Maria
M
Keating
Damien J
DJ
Chen
Chen
C
D000818:Animals; D001030:Apamin / Q000494:pharmacology; D045744:Cell Line, Tumor; D018999:Charybdotoxin / Q000494:pharmacology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000502:physiology; D006152:Cyclic GMP / Q000502:physiology*; D017869:Cyclic GMP-Dependent Protein Kinases / Q000502:physiology; D054439:Ghrelin; D013006:Growth Hormone / Q000378:metabolism; D036361:Peptide Hormones / Q000502:physiology*; D010902:Pituitary Gland; D024642:Potassium Channels, Voltage-Gated / Q000187:drug effects / Q000502:physiology*; D011493:Protein Kinase C / Q000502:physiology; D051381:Rats; D043562:Receptors, G-Protein-Coupled / Q000502:physiology; D054440:Receptors, Ghrelin; D015398:Signal Transduction / Q000502:physiology
D054439:Ghrelin; D036361:Peptide Hormones; D024642:Potassium Channels, Voltage-Gated; D043562:Receptors, G-Protein-Coupled; D054440:Receptors, Ghrelin; D018999:Charybdotoxin; D001030:Apamin; D013006:Growth Hormone; D017868:Cyclic AMP-Dependent Protein Kinases; D017869:Cyclic GMP-Dependent Protein Kinases; D011493:Protein Kinase C; D006152:Cyclic GMP