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• W2045210008 abstract "The phospholipase C (PLC)-activating gonadotrophin-releasing hormone (GnRH) receptor is thought not to rapidly desensitise in α T3-1 cells. This extremely unusual characteristic raises the concern that it might be a feature of the cell type, rather than the receptor per se. Here we have used video imaging to establish whether the effects of endogenous PLC-activating G-protein coupled receptors (GPCRs) on Ca 2+ ion concentration [Ca 2+ ] i desensitise in these cells. Oxytocin, endothelin-1, methacholine , and UTP all caused [Ca 2+ ] i increases which underwent rapid homologous desensitisation in that they were transient and responses to repeat stimuli were attenuated whereas subsequent responses to GnRH were not. To test whether receptor reserve obscures functional desensitisation of GnRH receptors, a photoaffinity antagonist (Pant-1), was used to effect a partial and irreversible receptor blockade . UV crosslinking in medium with 1000 nM Pant-1 reduced GnRH receptor number to 20±5% and reduced maximal buserelin-stimulated [ 3 H]IP X accumulation to 57±5%, demonstrating removal of receptor reserve. In control α T3-1 cells the initial rate of GnRH-stimulated [ 3 H]IP X accumulation was maintained for at least 5 min and GnRH caused a sustained increase in Ins(1,4,5)P 3 mass (confirming the resistance of GnRH receptors to desensitisation) and Pant-1 pre-treatment reduced the magnitude of these responses without altering their temporal profiles. In α T3-1 cells stably transfected with recombinant human muscarinic receptors ( α T3-1/M3), responses to methacholine were characteristic of desensitising GPCRs (transient Ins(1,4,5)P 3 and curvilinear [ 3 H]IP X responses) and were unaltered by Pant-1. To test the relevance of phospholipid pool size, α T3-1/M3 cells were pre-treated with GnRH or methacholine in medium with LiCl (to deplete PtdIns(4,5)P 2 pools). These pre-treatments reduced subsequent responses to methacholine and GnRH comparably, indicating access to a shared PtdIns(4,5)P 2 pool. Partial depletion of this pool (GnRH pre-treatment in medium with LiCl) reduced the magnitude of the [ 3 H]IP X and Ins(1,4,5)P 3 responses to methacholine and GnRH, without altering their temporal profiles. Thus the GnRH receptor does not undergo rapid homologous desensitisation in α T3-1 cells in spite of the fact that they can desensitise other endogenous (and recombinant) PLC-activating GPCRs, and the lack of desensitisation cannot be attributed to the existence of GnRH receptor reserve or access to an atypically large or rapidly re-cycled PtdIns(4,5)P 2 pool. This unique functional characteristic (mammalian GnRH receptors are the only PLC-activating GPCRs known not to rapidly desensitise) almost certainly therefore reflects the atypical structure of these receptors (mammalian GnRH receptors are the only PLC-activating GPCRs known to lack C-terminal tails)." @default.
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• W2045210008 date "1999-01-01" @default.
• W2045210008 modified "2023-09-26" @default.
• W2045210008 title "The lack of gonadotrophin-releasing hormone (GnRH) receptor desensitisation in αT3-1 cells is not due to GnRH receptor reserve or phosphatidylinositol 4,5-bis-phosphate pool size" @default.
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• W2045210008 doi "https://doi.org/10.1016/s0303-7207(98)00201-9" @default.
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