Perrard-Sapori M H, Chatelain P G, Jaillard C, Saez J M
Eur J Biochem. 1987 May 15;165(1):209-14. doi: 10.1111/j.1432-1033.1987.tb11213.x.
We have characterized somatomedin-C/insulin-like growth factor I (Sm-C/IGF-I) receptors in cultured pig Leydig cells by binding and cross-linking affinity experiments. At equilibrium the dissociation constant and the number of binding sites per cell were 1.8 +/- 0.2 X 10(-9) M and 12,200 +/- 3200 respectively. Under reducing conditions, disuccinimidyl suberate cross-linked 125I-Sm-C to one receptor complex with apparent Mr = 120,000. Continuous treatment of Leydig cells with increasing concentrations of human chorionic gonadotropin (hCG) for 48 h resulted in a dose-dependent (ED50 0.05 nM) increment in IGF type I receptors (2.5-3-fold increase). Conversely, treatment of Leydig cells for 48 h with increasing concentrations of Sm-C/IGF-I produced a 3-4-fold increase in hCG receptors. This effect was dose-dependent (ED50 = 7 ng/ml). Sm-C/IGF-I treatment also enhanced Leydig cell responsiveness to hCG for both cAMP (6-fold) and testosterone (8-fold). Moreover the stimulatory effects of Sm-C/IGF-I on cells cultured either in the absence or presence of 5% human serum from an hypopituitary patient were inhibited by anti-(Sm-C/IGF-I) antibodies. Taken together these results not only show that Leydig cells must be considered as targets for Sm-C/IGF-I, but also lend support to the possibility that Sm-C/IGF-I plays a role in the regulation of Leydig cell function. Moreover, they suggest that Sm-C/IGF-I might be responsible for the delayed puberty observed in some patients with isolated growth hormone deficiency.
我们通过结合和交联亲和力实验对培养的猪睾丸间质细胞中的生长调节素C/胰岛素样生长因子I(Sm-C/IGF-I)受体进行了表征。在平衡状态下,解离常数和每个细胞的结合位点数分别为1.8±0.2×10⁻⁹M和12,200±3200。在还原条件下,辛二酸二琥珀酰亚胺酯将¹²⁵I-Sm-C交联到一种表观分子量为120,000的受体复合物上。用浓度不断增加的人绒毛膜促性腺激素(hCG)连续处理睾丸间质细胞48小时,导致I型胰岛素样生长因子受体呈剂量依赖性增加(半数有效剂量为0.05 nM)(增加2.5至3倍)。相反,用浓度不断增加的Sm-C/IGF-I处理睾丸间质细胞48小时,导致hCG受体增加3至4倍。这种效应呈剂量依赖性(半数有效剂量=7 ng/ml)。Sm-C/IGF-I处理还增强了睾丸间质细胞对hCG产生环磷酸腺苷(6倍)和睾酮(8倍)的反应性。此外,抗(Sm-C/IGF-I)抗体抑制了Sm-C/IGF-I对在无垂体功能减退患者的5%人血清存在或不存在情况下培养的细胞的刺激作用。综上所述,这些结果不仅表明睾丸间质细胞必须被视为Sm-C/IGF-I的靶细胞,而且支持了Sm-C/IGF-I在调节睾丸间质细胞功能中起作用的可能性。此外,它们表明Sm-C/IGF-I可能是一些孤立性生长激素缺乏患者青春期延迟的原因。