Identification of beta-adrenoceptor subtypes in bovine ovarian and myometrial cell membranes

Beta-adrenoceptor (beta-AR) concentrations were measured in the ovary and in the myometrium of 36 adult Friesian cows using a radiometric assay. The beta-AR content in both tissues was determined using the highly specific antagonist (-) [3H]CGP 12177 and the amounts of beta-AR subtypes were discriminated in the presence of highly selective unlabelled ligands (CGP 20712A, ICI 118 551, CGP 25827A). Scatchard analysis revealed a good linearity and Kd values suggested the existence of high affinity beta-adrenergic sites in the bovine genital tract. Total beta-AR concentrations in the ovary and in the myometrium were, respectively, 87 +/- 7 (SEM) and 240 +/- 27 fmol mg-1 of membrane protein. beta 2-AR concentrations were significantly higher (P < 0.01) in the ovary (66 +/- 5) and the myometrium (180 +/- 29) than those of beta 1-AR (21 +/- 4 and 60 +/- 5, respectively). Significant differences (P < 0.05) were also to be found between high affinity state beta 2-AR and low affinity beta 2-AR concentrations, but their values correlated negatively in the two different tissues. Natural and synthetic agonists inhibited (-) [3H]CGP 12177 binding to beta-AR in the following order of potency: (-)isoproterenol > (+/-)clenbuterol > or = (-)adrenaline >> >> (-)noradrenaline, whereas synthetic antagonists inhibited binding in the following order of potency: (-)propranolol >> (+/-)ICI 118 551 >> >> (+/-)CGP 20712A.