BINDING OF LUTEINIZING HORMONE AND HUMAN CHORIONIC GONADOTROPHIN TO OVARIAN CELLS AND ACTIVATION OF ADENYLATE CYCLASE

Abstract

SUMMARY The binding of 125I-labelled ovine luteinizing hormone (OLH) and human chorionic gonadotrophin (HCG) to unbroken cells or homogenates from rat and bovine ovaries was studied in relation to the hormonal activation of adenylate cyclase. Both hormones are preferentially bound to ovarian tissue (luteal as well as follicular), while adrenal, kidney and liver showed no specific binding. The bound HCG was associated chiefly with the fraction of the homogenate sedimenting at 1000 g. Addition of excess unlabelled hormone (HCG, OLH) together with the corresponding or heterologous 125I-labelled hormone reduced the radioactivity bound to the tissue by 70–90%; rat LH also competed with OLH and HCG for binding sites, but follicle-stimulating hormone, prolactin, growth hormone, adrenocorticotrophin or glucagon were without effect. Binding could be dissociated from activation of adenylate cyclase by a two-stage incubation: at 4 °C rat ovarian slices bound the labelled hormones, though binding efficiency was reduced to about 20%, but no stimulation of the enzyme was observed. Reincubation of these slices, after washing, at 37 °C in hormone-free medium resulted in maximal activation of adenylate cyclase. Addition of homologous antibodies before the second incubation step abolished the effect of OLH but not that of HCG on the activation of the enzyme. Similarly, addition of homologous antibodies or of excess unlabelled hormone caused dissociation of bound 125I-labelled OLH but not of 125I-labelled HCG from receptor sites. It is concluded that (i) HCG, OLH and rat LH are bound to the same receptor on ovarian cells, but binding of OLH to the receptor is more labile than that of HCG; (ii) the receptor appears to be located on the plasma membrane, since it was largely associated with the 1000 g pellet and since bound OLH remained accessible to antibody; (iii) activation of adenylate cyclase requires the continued occupation of the binding site by the hormone, but occupation of only a fraction of the receptor sites by hormone is adequate to induce maximal production of cyclic AMP by the target tissue.