Characteristics of the Ca2+ spike and oscillations induced by different doses of thyrotropin-releasing hormone (TRH) in individual pituitary cells and nonexcitable cells transfected with TRH receptor complementary deoxyribonucleic acid.
The Ca2+ response of individual cells to TRH was investigated in excitable pituitary GH3 and in nonexcitable Hela cells transfected with the TRH receptor complementary DNA (HelaR cells). GH3 cells typically responded to 1 microM TRH with an immediate transient [Ca2+]i spike (mean peak [Ca2+]i = 1.5 microM) followed by a period of inactivity of approximately 100 sec long and then a secondary increase in [Ca2+]i with oscillations. At 10-100 nM TRH, the initial [Ca2+]i spike was more prolonged and immediately followed by a sustained elevation of [Ca2+]i. At 0.5-1 nM TRH, there was a variable lag before any response; the initial [Ca2+]i spike was absent or small, but the sustained phase was still present. The second phase of elevated [Ca2+]i, which could be eliminated with nimodipine or chelation of extracellular Ca2+, gave a bell-shaped TRH dose response curve. The effect of TRH on Ca2+ oscillations depended both on TRH concentration and the basal oscillation frequency. HelaR cells responded to 1 microM TRH with a rapid [Ca2+]i spike, and at less than or equal to 10 nM TRH, up to 50% of HelaR cells displayed agonist-induced sinusoidal [Ca2+]i oscillations independent of extracellular Ca2+. TRH never caused a sustained elevation of [Ca2+]i in HelaR cells. For GH3 and HelaR cells, the peak [Ca2+]i response increased with TRH concentration up to 1 microM. In contrast, the duration of the initial [Ca2+]i spike was shorter at higher TRH concentrations, decreasing from 16 to 6.3 s (GH3) or 92 to 35 s (HelaR) between 0.5 nM and 1 microM TRH. This shortening of the spike duration was caused by rapid clearing of cytoplasmic Ca2+ that depended primarily on agonist concentration. In summary, TRH stimulates a complex [Ca2+]i response pattern dependent upon both the agonist concentration and cell context. The initial burst of Ca2+ is cleared in part by agonist dependent Ca2+ clearing.
Nelson
E J
EJ
Department of Pharmacology, University of Rochester School of Medicine and Dentistry, New York 14642.
Hinkle
P M
PM
D000818:Animals; D002118:Calcium / Q000502:physiology*; D002460:Cell Line; D018076:DNA, Complementary; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D006367:HeLa Cells / Q000187:drug effects* / Q000502:physiology; D006801:Humans; D007425:Intracellular Membranes / Q000378:metabolism; D009991:Oscillometry; D009994:Osmolar Concentration; D010902:Pituitary Gland / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D051381:Rats; D018025:Receptors, Thyrotropin-Releasing Hormone / Q000235:genetics*; D013973:Thyrotropin-Releasing Hormone / Q000494:pharmacology*; D014162:Transfection
D018076:DNA, Complementary; D018025:Receptors, Thyrotropin-Releasing Hormone; D013973:Thyrotropin-Releasing Hormone; D002118:Calcium