Counis R, Dufour S, Ribot G, Quérat B, Fontaine Y A, Jutisz M
Endocrinology. 1987 Sep;121(3):1178-84. doi: 10.1210/endo-121-3-1178.
In teleosts, the pituitary contains a single glycoprotein gonadotropic hormone (GTH) composed of two dissimilar alpha- and beta-subunits. The European eel, Anguilla anguilla L, is sexually immature at the silver stage due to a deficiency in GTH synthesis and secretion. In previous studies we (S.D., YA.F.) have demonstrated a strong stimulatory action of estradiol (E2) on eel pituitary GTH content. In contrast, we (R.C., M.J.) have shown that in the rat E2 negatively regulates gonadotropin subunit synthesis via changes in specific mRNA levels. The purpose of our present work was to check for such effects of E2 on the synthesis of GTH alpha- and beta-subunits in the European eel. Eel pituitary mRNA was translated in a cell-free system in the presence of [35S]Met + Cys. We demonstrate that one of the translated polypeptides, characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and fluorography, cross-reacts with an antiserum to denatured bovine alpha-subunit. Its apparent mol wt (18.5K), which is slightly higher than that of the corresponding rat alpha-precursor, suggests that it represents the precursor of the alpha-subunit of eel glycoprotein hormones. The specificity of immunoprecipitation was confirmed by competition with ovine alpha (but not with ovine LH beta or bovine TSH beta). Quantitative evaluation of the putative eel alpha-subunit precursor showed that it represents 0.2% of the total protein translated by RNA from the normal silver eel. Chronic treatment of eels for 3 weeks with 17 beta-E2 increased by 8.0- to 8.5-fold the proportion of the putative alpha-subunit precursor among translation products. Due to the lack of cross-reactivity with the presumed GTH beta precursor, no radioactive material could be specifically detected in translation medium of eel pituitary mRNA using antisera to either denatured bovine LH beta or ovine FSH beta. Our data suggest that E2, depending on vertebrate group and probably on sexual status, may exert either positive or negative control on gonadotropin synthesis by opposite effects on the levels of specific mRNA.
在硬骨鱼类中,脑垂体含有一种单一的糖蛋白促性腺激素(GTH),它由两个不同的α-和β-亚基组成。欧洲鳗鲡(Anguilla anguilla L)在银色阶段性未成熟,这是由于GTH合成和分泌不足所致。在先前的研究中,我们(S.D., YA.F.)已证明雌二醇(E2)对鳗鲡脑垂体GTH含量有强烈的刺激作用。相反,我们(R.C., M.J.)已表明,在大鼠中,E2通过特定mRNA水平的变化对促性腺激素亚基合成起负调节作用。我们目前这项工作的目的是检查E2对欧洲鳗鲡GTHα-和β-亚基合成的此类影响。鳗鲡脑垂体mRNA在存在[35S]甲硫氨酸 + 半胱氨酸的无细胞系统中进行翻译。我们证明,通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳和荧光自显影表征的一种翻译多肽与抗变性牛α-亚基抗血清发生交叉反应。其表观分子量(18.5K)略高于相应大鼠α-前体的分子量,这表明它代表鳗鲡糖蛋白激素α-亚基的前体。免疫沉淀的特异性通过与羊α(但不与羊促黄体生成素β或牛促甲状腺激素β)竞争得以证实。对假定的鳗鲡α-亚基前体的定量评估表明,它占正常银色鳗鲡RNA翻译的总蛋白的0.2%。用17β-E2对鳗鲡进行3周的慢性处理,使假定的α-亚基前体在翻译产物中的比例增加了8.0至8.5倍。由于与假定的GTHβ前体缺乏交叉反应,使用抗变性牛促黄体生成素β或羊促卵泡激素β抗血清在鳗鲡脑垂体mRNA的翻译介质中无法特异性检测到放射性物质。我们的数据表明,E2根据脊椎动物类别以及可能的性状态,可能通过对特定mRNA水平的相反作用对促性腺激素合成施加正调控或负调控。