Different G proteins are involved in the biphasic response of clonal rat pituitary cells to thyrotropin-releasing hormone.
In rat anterior pituitary tumour cells (GH3/B6) thyrotropin-releasing hormone (TRH) elicits a biphasic response. First, a release of intracellularly stored Ca2+ induces a hyperpolarization of the cell. Second, a depolarization thought to be induced by a reduction of the inward-rectifying K+ current (KIR) causes an increase in action potential frequency and a plateau-like increase in [Ca2+]i. It has been proposed that the two phases are induced by the actions of inositol 1,4,5-trisphosphate (InsP3) and protein kinase C (PKC), respectively, but we demonstrate here that PKC is not responsible for the second phase increase in [Ca2+]i and suggest that the pathways diverge at the level of receptor and G protein coupling. Both phases of the TRH response were insensitive to pertussis toxin, but cholera toxin (CTX) selectively affected the second phase. After CTX pretreatment cells had a high spontaneous spiking frequency and smaller KIR amplitude. In these cells TRH failed to increase the action potential frequency after the first phase hyperpolarization, elicited only a transient peak increase in [Ca2+]i with no plateau phase and could only slightly reduce KIR. These effects of CTX are not mediated by its ability to increase cAMP via activation of GS, as increased cAMP levels neither inhibit KIR nor prevent its reduction by TRH. In addition, inhibition of protein kinase A activation did not block the second phase increase in [Ca2+]i induced by TRH, suggesting that the CTX-sensitive G protein mediating the second phase of the TRH response is not GS.
Bauer
C K
CK
Physiologisches Institut, UKE, Hamburg, Germany.
Davison
I
I
Kubasov
I
I
Schwarz
J R
JR
Mason
W T
WT
D000200:Action Potentials / Q000187:drug effects; D000818:Animals; D002478:Cells, Cultured; D002772:Cholera Toxin / Q000494:pharmacology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D019204:GTP-Binding Proteins / Q000502:physiology*; D007091:Image Processing, Computer-Assisted; D009761:Nystatin; D018408:Patch-Clamp Techniques; D037342:Pertussis Toxin; D010902:Pituitary Gland / Q000166:cytology / Q000502:physiology*; D015221:Potassium Channels / Q000187:drug effects / Q000378:metabolism; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D051381:Rats; D013973:Thyrotropin-Releasing Hormone / Q000494:pharmacology*; D010566:Virulence Factors, Bordetella / Q000494:pharmacology
D015221:Potassium Channels; D010566:Virulence Factors, Bordetella; D009761:Nystatin; D013973:Thyrotropin-Releasing Hormone; D002772:Cholera Toxin; D037342:Pertussis Toxin; D017868:Cyclic AMP-Dependent Protein Kinases; D011493:Protein Kinase C; D019204:GTP-Binding Proteins