In-vitro binding of gonadotrophin to fish ovary.
Binding of piscine and mammalian gonadotrophin to plasma membranes from the ovaries of a fish, the murrel (Channa punctatus), clearly suggests that the fish ovary possesses distinct and specific binding sites for both piscine and mammalian gonadotrophins. Maximum specific binding of 125I-labelled human chorionic gonadotrophin (125I-hCG) and 125I-labelled silver carp gonadotrophin (125I-scG) was obtained at 30 degrees C and pH 7.5 during 2 h of incubation. In competitive binding studies, binding of radiolabelled scG was effectively inhibited by piscine gonadotrophins while LH and hCG had less effect and FSH showed no inhibition. By using plasma membrane preparations from kidney, skeletal muscle, brain and ovary it could be shown that specific binding of radiolabelled gonadotrophins was restricted to ovarian tissue. Binding characteristics of both 125I-scG and 125I-hCG to a preparation of murrel ovarian plasma membranes showed saturability with high affinity and low capacity. Scatchard plot analysis gave a higher dissociation constant for hCG (Kd = 235 pmol/l) than for scG (Kd = 127 pmol/l). Maximum binding capacity of scG was about twofold higher (6.27 fmol/mg protein) than that of hCG (3.76 fmol/mg protein). An increase in gonadotrophin binding resulted in a greater formation of pregnenolone from cholesterol, indicating functional relevance. At a concentration of 8 mmol/l, Ca2+ markedly inhibited the binding of gonadotrophin. The physiological importance of this inhibition is discussed.
D000818:Animals; D001665:Binding Sites / Q000187:drug effects; D002413:Cations, Divalent / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D006063:Chorionic Gonadotropin / Q000378:metabolism; D005260:Female; D005399:Fishes / Q000378:metabolism*; D006062:Gonadotropins / Q000378:metabolism*; D066298:In Vitro Techniques; D009865:Oocytes / Q000378:metabolism; D010053:Ovary / Q000378:metabolism*
D002413:Cations, Divalent; D006063:Chorionic Gonadotropin; D006062:Gonadotropins