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. 1990 Jun 1;268(2):345–352. doi: 10.1042/bj2680345

Thyrotropin-releasing hormone activates a [Ca2+]i-dependent K+ current in GH3 pituitary cells via Ins(1,4,5)P3-sensitive and Ins(1,4,5)P3-insensitive mechanisms.

P Mollard 1, B Dufy 1, P Vacher 1, J L Barker 1, W Schlegel 1
PMCID: PMC1131438  PMID: 2163608

Abstract

The role of Ins(1,4,5)P3 in receptor-induced Ca2+ mobilization in pituitary cells was studied at the single-cell level. Experimental strategies were developed which allowed a comparative analysis of the effects of Ins(1,4,5)P3 with those of receptor activation under identical conditions. These include microfluorimetry as well as a novel technique which permits the controlled and rapid application of intracellular messenger molecules to individual cells. This latter approach is based on the tight-seal whole-cell recording (WCR) technique, and utilizes two patch-clamp micropipettes, one for electrical recording and the second for the controlled pressure injection. Ins(1,4,5)P3, when applied with this dual-WCR (DWCR) technique, leads rapidly to a marked rise in cytosolic free Ca2+ [( Ca2+]i) and a concomitant stimulation of Ca2(+)-activated K+ current; Ins(1,4,5)P3 can thus mimic the effects of thyrotropin-releasing hormone (TRH) in the same cells under identical conditions. In cells dialysed intracellularly with heparin, a potent antagonist of Ins(1,4,5)P3 action, the rapid response to extracellular stimulation with TRH was abolished, as were the effects of intracellular application of Ins(1,4,5)P3. Heparin, which abolished Ins(1,4,5)P3 action completely, blocked responses to TRH in some cells only partially, revealing that Ca2+ mobilization response to TRH is in part slower in onset than the response to Ins(1,4,5)P3. It is concluded (1) that Ins(1,4,5)P3 is an essential element for the action of TRH, providing a rapid mechanism for Ca2+ mobilization induced by the releasing hormone and (2) that TRH action in mobilizing intracellular Ca2+ is sustained by a slower mechanism which is independent of Ins(1,4,5)P3.

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Selected References

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