Thyrotropin-releasing hormone (TRH) elevation of inositol trisphosphate and cytosolic free calcium is dependent on receptor number. Evidence for multiple rapid interactions between TRH and its receptor.
Thyrotropin-releasing hormone (TRH) increases rapidly two potential intracellular signals, inositol trisphosphate (IP3) and free cytosolic calcium ([Ca2+]i), for stimulated prolactin release and synthesis in GH4C1 rat pituitary cells. We have examined the temporal relationships between TRH-enhanced formation of inositol phosphates and TRH-elevated [Ca2+]i. TRH-enhanced IP3 content was closely paralleled by the initial phase of TRH-elevated [Ca2+]i. To investigate receptor-effector coupling for these rapid actions of TRH, we examined their dependence on receptor number in five GH4C1 variant strains containing 0-2.6 X 10(5) receptor sites/cell. We found that receptor number (up to 1.7 X 10(5)/cell) was limiting for TRH-enhanced IP3 formation as well as for both the initial burst and plateau phases of TRH-elevated [Ca2+]i. The ED50 for rapid (5 s) TRH-stimulated IP3 formation was higher than for other sustained TRH actions in these cells, and we postulated that the initial TRH receptor interactions occur with rapid dissociation kinetics. To test this hypothesis, we performed rapid dilution experiments following a 1-s stimulation and found that TRH-stimulated IP3 formation decreased within 4 s of dilution and disappeared within 60 s at which time fresh TRH could restimulate IP3 formation. We conclude that receptor occupancy is the limiting step for TRH-stimulated IP3 formation and elevated [Ca2+]i and that maximal TRH action requires multiple rapid interactions between TRH and its receptor.
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D015544:Inositol 1,4,5-Trisphosphate; D007295:Inositol Phosphates / Q000378:metabolism*; D007700:Kinetics; D010911:Pituitary Neoplasms / Q000378:metabolism; D051381:Rats; D011956:Receptors, Cell Surface / Q000378:metabolism*; D018025:Receptors, Thyrotropin-Releasing Hormone; D013403:Sugar Phosphates / Q000378:metabolism*; D013973:Thyrotropin-Releasing Hormone / Q000494:pharmacology*; D013997:Time Factors
D007295:Inositol Phosphates; D011956:Receptors, Cell Surface; D018025:Receptors, Thyrotropin-Releasing Hormone; D013403:Sugar Phosphates; D013973:Thyrotropin-Releasing Hormone; D015544:Inositol 1,4,5-Trisphosphate; D002118:Calcium