title: Signalling, cycling and desensitisation of gonadotrophin-releasing hormone receptors.
abstract: Sustained stimulation of G-protein-coupled receptors (GPCRs) typically causes receptor desensitisation, which is mediated by phosphorylation, often within the C-terminal tail of the receptor. The consequent binding of beta-arrestin not only prevents the receptor from activating its G protein (causing desensitisation), but can also target it for internalisation via clathrin-coated vesicles and can mediate signalling to proteins regulating endocytosis and mitogen-activated protein kinase (MAPK) cascades. GnRH acts via phospholipase C (PLC)-coupled GPCRs on pituitary gonadotrophs to stimulate a Ca(2+)-mediated increase in gonadotrophin secretion. The type I GnRH receptors (GnRH-Rs), found only in mammals, are unique in that they lack C-terminal tails and apparently do not undergo agonist-induced phosphorylation or bind beta-arrestin; they are therefore resistant to receptor desensitisation and internalise slowly. In contrast, the type II GnRH-Rs, found in numerous vertebrates, possess such tails and show rapid desensitisation and internalisation, with concomitant receptor phosphorylation (within the C-terminal tails) or binding of beta-arrestin, or both. The association with beta-arrestin may also be important for regulation of dynamin, a GTPase that controls separation of endosomes from the plasma membrane. Using recombinant adenovirus to express GnRH-Rs in Hela cells conditionally expressing a dominant negative mutant of dynamin (K44A), we have found that blockade of dynamin-dependent endocytosis inhibits internalisation of type II (xenopus) GnRH-Rs but not type I (human) GnRH-Rs. In these cells, blockade of dynamin-dependent internalisation also inhibited GnRH-R-mediated MAPK activation, but this effect was not receptor specific and therefore not dependent upon dynamin-regulated GnRH-R internalisation. Although type I GnRH-Rs do not desensitise, sustained activation of GnRH-Rs causes desensitisation of gonadotrophin secretion, and we have found that GnRH can cause down-regulation of inositol (1,4,5) trisphosphate receptors and desensitisation of Ca(2+) mobilisation in pituitary cells. The atypical resistance of the GnRH-R to desensitisation may underlie its atypical efficiency at provoking this downstream adaptive response. GnRH-Rs are also expressed in several extrapituitary sites, and these may mediate direct inhibition of proliferation of hormone-dependent cancer cells. Infection with type I GnRH-R-expressing adenovirus facilitated expression of high-affinity, PLC-coupled GnRH-R in mammary and prostate cancer cells, and these mediated pronounced antiproliferative effects of receptor agonists. No such effect was seen in cells transfected with a type II GnRH-R, implying that it is mediated most efficiently by a non-desensitising receptor. Thus it appears that the mammalian GnRH-Rs have undergone a period of rapidly accelerated molecular evolution that is of functional relevance to GnRH-Rs in pituitary and extrapituitary sites.
authors.affiliation: University Research Centre for Neuroendocrinology, University of Bristol, Bristol BS2 8HW, UK. craig.mcardle@bristol.ac.uk
authors.forename: Craig A
authors.initials: CA
authors.lastname: McArdle
authors.forename: J
authors.initials: J
authors.lastname: Franklin
authors.forename: L
authors.initials: L
authors.lastname: Green
authors.forename: J N
authors.initials: JN
authors.lastname: Hislop
mesh_terms: D000818:Animals; D019390:Arrestins / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology; D019143:Evolution, Molecular; D007987:Gonadotropin-Releasing Hormone / Q000378:metabolism*; D006801:Humans; D008322:Mammals / Q000378:metabolism*; D009369:Neoplasms / Q000378:metabolism; D010902:Pituitary Gland / Q000378:metabolism*; D011485:Protein Binding; D011966:Receptors, LHRH / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction / Q000502:physiology*; D020079:Terminal Repeat Sequences; D000071557:beta-Arrestins
chemical_list: D019390:Arrestins; D011966:Receptors, LHRH; D000071557:beta-Arrestins; D007987:Gonadotropin-Releasing Hormone