Bonnin J P
Laboratoire de Biologie marine, Université de Bordeaux-I, Talence.
C R Acad Sci III. 1989;309(10):435-40.
Cultured seminal vesicle cells of Gobius niger L., precultured for about 15 days, are tested for their Testosterone-binding capacity. This whole cell system shows a specific binding of the androgen, reaching saturation in the presence of increasing amounts of ligand and the Scatchard Plot indicates a good affinity (KD = 4.4 x 10(-9) M), involving a number of sites of 5.06 x 10(-15) mole/culture. The possible existence of a second binding-site population with lower affinity and greater number of sites remains to be demonstrated. At 18 degrees C, the time course shows a maximal binding after about 60 min. of incubation, followed by a rapid decline at 90 min. The competition experiments involving estradiol, 11-keto- and 11-hydroxytestosterone indicate an effective if not total specificity to these steroids. The cross-reaction percentages at the 50% binding level are respectively 10.9, 8.5, 4.02%. Ovine Prolactin treatment of the cultures for 6 days before the binding experiment significantly improves the specific binding level of testosterone if compared to the controls (p .019, p .005). This result indicates that the Prolactin-Testosterone synergistic data already obtained on the seminal vesicles of Siluridae and Gobiidae is explained by a direct effect of the Prolactin on the androgen receptor level in the seminal vesicle cell.
对预先培养约15天的黑腹栉虾虎鱼精囊细胞进行睾酮结合能力测试。这个完整细胞系统显示出雄激素的特异性结合,在配体数量增加时达到饱和,Scatchard图表明具有良好的亲和力(KD = 4.4×10⁻⁹ M),涉及的位点数量为5.06×10⁻¹⁵摩尔/培养物。亲和力较低但位点数量较多的第二个结合位点群体的可能存在仍有待证明。在18℃下,时间进程显示孵育约60分钟后结合达到最大值,随后在90分钟时迅速下降。涉及雌二醇、11-酮睾酮和11-羟基睾酮的竞争实验表明,对这些类固醇具有有效的(即使不是完全的)特异性。在50%结合水平下的交叉反应百分比分别为10.9%、8.5%、4.02%。在结合实验前对培养物进行6天的绵羊催乳素处理,与对照组相比,显著提高了睾酮的特异性结合水平(p <.019,p <.005)。这一结果表明,已经在鲶科和虾虎鱼科精囊上获得的催乳素-睾酮协同作用数据可以通过催乳素对精囊细胞中雄激素受体水平的直接作用来解释。