Breton B, Le Gac F, Sambroni E
Gen Comp Endocrinol. 1986 Nov;64(2):163-71. doi: 10.1016/0016-6480(86)90001-8.
A method has been developed suitable for the study of the binding of salmon (Oncorhynchus tsawytscha) gonadotropic hormone (GtH) to brown trout (Salmo trutta) ovarian particulate fractions. Several proteolytic enzyme inhibitors were tested to preserve the binding capacity of particulate fractions (1000, 20,000, and 110,000g) from yolky oocytes. The best preservation was obtained with trypsin inhibitor. The effects on binding of the pH, time, and temperature of the incubation were investigated. Binding increased linearly with amount of particulate fraction, and is saturable. In both the 20,000 and 110,000g fractions, there are two classes of receptors: one with a high-affinity constant (3.14 and 0.95 10(10) M-1 for the 20,000 and 110,000g fractions, respectively) and a low capacity (1.76 and 0.63 fmol/g of ovary), the other with a higher-affinity constant (3.44 and 1.78 10(9) M-1) but a higher capacity (7.37 and 7.42 fmol/g of ovary). GtH binding was not affected by ovine luteinizing hormone (oHL), ovine follice-stimulating hormone (oFSH), and human chorionic gonadotropin (hCG), but was partially inhibited by 1 micrograms of bovine thyroid-stimulating hormone (bTSH) and salmon prolactin (sPRL). The results are discussed in relation to the biological specificity of fish GtH and to the unsolved problem concerning the number of gonadotropins in fish.
已开发出一种适用于研究鲑鱼（虹鳟）促性腺激素（GtH）与褐鳟卵巢微粒体部分结合的方法。测试了几种蛋白水解酶抑制剂以保留来自有卵黄卵母细胞的微粒体部分（1000、20000和110000g）的结合能力。用胰蛋白酶抑制剂可获得最佳保存效果。研究了孵育的pH、时间和温度对结合的影响。结合随微粒体部分的量呈线性增加，且具有饱和性。在20000和110000g的部分中，有两类受体：一类具有高亲和力常数（20000g和110000g部分分别为3.14和0.95×10¹⁰M⁻¹）和低容量（分别为1.76和0.63fmol/g卵巢），另一类具有更高的亲和力常数（3.44和1.78×10⁹M⁻¹）但更高的容量（7.37和7.42fmol/g卵巢）。GtH结合不受绵羊促黄体激素（oHL）、绵羊促卵泡激素（oFSH）和人绒毛膜促性腺激素（hCG）的影响，但受到1微克牛促甲状腺激素（bTSH）和鲑鱼催乳素（sPRL）的部分抑制。结合鱼类GtH的生物学特异性以及鱼类促性腺激素数量方面尚未解决的问题对结果进行了讨论。
