Féral C, Benhaïm A, Leymarie P
Laboratoire de Biochimie, EP CNRS 9, CHU Côte de Nacre, Caen, France.
Biochim Biophys Acta. 1996 Oct 23;1284(2):221-6. doi: 10.1016/s0005-2736(96)00134-4.
Specific, high-affinity angiotensin II (A II) receptors were observed on granulosa and thecal cells of preovulatory ovarian follicles from immature PMSG-treated rabbits. Scatchard analysis of 125I-[Sar1,Ile8]A II binding to freshly prepared cells was indicative of only one class of binding sites. Kd values were 0.26 +/- 0.11 nM and 0.18 +/- 0.02 nM, densities of A II receptors were 0.06 +/- 0.02 fmol/10(5) cells and 0.08 +/- 0.01 fmol/10(5) cells for granulosa and thecal cells, respectively. When cells were incubated for 48 h with hCG, Kd values were of the same order of magnitude, but the amount of A II receptors was increased 2-fold in granulosa and 4-fold in theca. Using subtype specific ligands (Losartan for AT1 and PD 123319 for AT2) in competitive binding experiments, A II receptors were found to be of the AT1 type on both granulosa and thecal cells freshly prepared or incubated 48 h in vitro. These results establishing the existence of high affinity AT1 receptors on the two cell types of the rabbit preovulatory follicles contrast with previous observations showing the presence of AT2 receptors on granulosa or theca from several species.
在未成熟的经孕马血清促性腺激素(PMSG)处理的兔排卵前卵巢卵泡的颗粒细胞和卵泡膜细胞上观察到了特异性、高亲和力的血管紧张素II(A II)受体。对新鲜制备的细胞进行125I-[Sar1,Ile8]A II结合的Scatchard分析表明只有一类结合位点。颗粒细胞和卵泡膜细胞的解离常数(Kd)值分别为0.26±0.11 nM和0.18±0.02 nM,A II受体密度分别为0.06±0.02 fmol/10(5)个细胞和0.08±0.01 fmol/10(5)个细胞。当细胞与hCG孵育48小时后,Kd值处于相同数量级,但颗粒细胞中A II受体的量增加了2倍,卵泡膜细胞中增加了4倍。在竞争性结合实验中使用亚型特异性配体(氯沙坦用于AT1,PD 123319用于AT2),发现新鲜制备的或体外孵育48小时的颗粒细胞和卵泡膜细胞上的A II受体均为AT1型。这些结果证实了兔排卵前卵泡的两种细胞类型上存在高亲和力的AT1受体,这与之前在几种物种的颗粒细胞或卵泡膜细胞上发现AT2受体的观察结果形成了对比。