Freidman S, Gurevich M, Shemesh M
Department of Hormone Research, Koret School of Veterinary Medicine Hebrew University, Bet Dagan, Israel.
Biol Reprod. 1995 May;52(5):1020-6. doi: 10.1095/biolreprod52.5.1020.
High-affinity LH/hCG binding sites have been characterized in porcine, lepine, and murine uteri. In the present study, LH/hCG binding sites were characterized in bovine endometrium. Radioreceptor assays were performed with membrane homogenates of endometrial tissues and analyzed for binding site specificity and capacity. There was little competition for receptor occupancy between hCG and ovine FSH (5%) or ovine prolactin (< 0.1%), but there was a 20% cross-reaction with eCG. There was no affinity for LH/hCG in crude membrane preparations of kidney, skeletal muscle, or vascular tissues. Concentrations of endometrial LH/hCG binding sites were determined during the bovine estrous cycle. LH/hCG receptors were found in cell preparations from Days 2-4 and 15-17 of the cycle, but not in preparations from the other stages of the cycle tested (Days 8-12, pre- and post-estrus, and ovulation). The concentration of uterine LH/hCG receptor varied during the estrous cycle, with higher values at Days 15-17 (3.1 fmol/mg protein) and lower values at Days 2-4 (1.2 fmol/mg protein). However, the binding capacity of hCG by luteal cells (9.7 fmol/mg protein) was 3-fold higher (p < 0.01) than that by endometrial tissue on any day studied. No differences in affinity constant (Ka) were seen between endometrial LH/hCG receptors (either) from Days 2-4 or 15-17) and mid-cycle luteal cells (0.60 x 10(11) M-1). Using Western blot analysis, we determined the expression of cyclooxygenase (COX) during the estrous cycle of the cow. It was found that the signal for COX was strongest at 15-17 days.(ABSTRACT TRUNCATED AT 250 WORDS)
高亲和力的促黄体生成素/人绒毛膜促性腺激素(LH/hCG)结合位点已在猪、兔和鼠的子宫中得到表征。在本研究中,对牛子宫内膜中的LH/hCG结合位点进行了表征。采用子宫内膜组织的膜匀浆进行放射受体分析,并分析结合位点的特异性和容量。hCG与羊促卵泡素(FSH,5%)或羊催乳素(<0.1%)之间对受体占据的竞争很小,但与马绒毛膜促性腺激素(eCG)有20%的交叉反应。在肾脏、骨骼肌或血管组织的粗膜制剂中,对LH/hCG没有亲和力。在牛发情周期中测定了子宫内膜LH/hCG结合位点的浓度。在发情周期的第2 - 4天和15 - 17天的细胞制剂中发现了LH/hCG受体,但在测试的周期其他阶段(第8 - 12天、发情前后和排卵)的制剂中未发现。子宫LH/hCG受体的浓度在发情周期中有所变化,在第15 - 17天较高(3.1飞摩尔/毫克蛋白质),在第2 - 4天较低(1.2飞摩尔/毫克蛋白质)。然而,在任何研究日,黄体细胞对hCG的结合能力(9.7飞摩尔/毫克蛋白质)比子宫内膜组织高3倍(p < 0.01)。在第2 - 4天或15 - 17天的子宫内膜LH/hCG受体与周期中期黄体细胞(0.60×10¹¹ M⁻¹)之间,未观察到亲和力常数(Ka)的差异。使用蛋白质免疫印迹分析,我们测定了母牛发情周期中环氧合酶(COX)的表达。发现COX的信号在15 - 17天时最强。(摘要截短于250字)