Mechanism of membrane electrical response to thyrotropin-releasing hormone in Xenopus oocytes injected with GH3 pituitary cell messenger ribonucleic acid.
TRH evoked a complex electrical membrane response in Xenopus laevis oocytes injected with either total cytosolic or poly(A)(+)-enriched RNA from GH3 pituitary cells but not in uninjected oocytes. A typical response consisted of a transient, rapid depolarizing current followed by a prolonged depolarizing current with superimposed current fluctuations. The reversal potentials of the rapid and the slow components of the response were -23.0 and -22.6 mV, respectively, and were markedly affected by CI- concentration indicating that the TRH response was mainly an increase in Cl- conductance. The response to TRH was dose dependent and was inhibited by the TRH antagonist, chlordiazepoxide. TRH caused rapid hydrolysis of labeled phosphatidylinositol 4,5-bisphosphate and a marked, prolonged increase in 45Ca2+ efflux from injected oocytes. The depolarizing response to TRH was not diminished in oocytes incubated in a Ca2(+)-free medium, but was inhibited by microinjection of EGTA. These data suggest that TRH evokes an electrophysiological response in oocytes injected with RNA from GH3 cells via activation of the same biochemical pathway that mediates its actions in GH3 cells. This pathway involves hydrolysis of phosphatidylinositol 4,5-bisphosphate, forming inositol trisphosphate that causes mobilization of cellular Ca2+. We suggest that oocytes injected with GH3 cell RNA, because of their large size and easy access to their intracellular milieu, will be a useful intact cell model in which to define the molecular details of signal transduction by TRH.
Oron
Y
Y
Department of Medicine, Cornell University Medical College, New York, New York 10021.
Gillo
B
B
Straub
R E
RE
Gershengorn
M C
MC
D000818:Animals; D002118:Calcium / Q000378:metabolism; D006868:Hydrolysis; D015544:Inositol 1,4,5-Trisphosphate / Q000378:metabolism; D008055:Lipids / Q000032:analysis; D008564:Membrane Potentials / Q000187:drug effects; D009865:Oocytes / Q000187:drug effects*; D019269:Phosphatidylinositol 4,5-Diphosphate; D010716:Phosphatidylinositols / Q000378:metabolism; D010911:Pituitary Neoplasms / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D012334:RNA, Neoplasm / Q000302:isolation & purification; D013973:Thyrotropin-Releasing Hormone / Q000494:pharmacology*; D014407:Tumor Cells, Cultured; D014982:Xenopus laevis
D008055:Lipids; D019269:Phosphatidylinositol 4,5-Diphosphate; D010716:Phosphatidylinositols; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D013973:Thyrotropin-Releasing Hormone; D015544:Inositol 1,4,5-Trisphosphate; D002118:Calcium