Maitre J L, Mercier L, Dolo L, Valotaire Y
Biochimie. 1985 Feb;67(2):215-25. doi: 10.1016/s0300-9084(85)80050-x.
In all egg laying vertebrates, synthesis and use of vitellogenin (Vg) are intimately bound to the active phase of reproduction. In the liver of the rainbow trout (Salmo gairdnerii), Vg synthesis is influenced by estradiol (E2) which, we believe, acts through the classical mechanism of steroid hormone action. After binding of the hormone to a soluble specific receptor protein, the estradiol-receptor complex can interact with chromatin and modulate the expression of Vg genes, leading to increased synthesis of specific mRNA and Vg. We show here: (i) the presence of specific oestrogen receptors (dissociation constant KD congruent to 1.5 X 10(-9) M for E2) in the cytosol of the male trout liver. (ii) The male liver, offering, an ideal experimental control of "zero" background, we followed-in the liver of male trout--the kinetics of induction of Vg mRNA by hybridization with Vg cDNA, after E2 stimulation, and (iii) the apparition of Vg in the serum by using an original rocket immuno-electrophoretic technique. The male trout liver vitellogenin model and the original techniques we developed will be very useful to study the influence of endogenous and exogenous factors on the different steps (receptors, transcription, translation) of vitellogenesis regulation.
在所有产卵脊椎动物中,卵黄蛋白原(Vg)的合成与利用都与生殖活跃期密切相关。在虹鳟鱼(Salmo gairdnerii)的肝脏中,Vg的合成受雌二醇(E2)影响,我们认为E2是通过类固醇激素作用的经典机制发挥作用的。激素与可溶性特异性受体蛋白结合后,雌二醇 - 受体复合物可与染色质相互作用并调节Vg基因的表达,导致特定mRNA和Vg的合成增加。我们在此展示:(i)雄性鳟鱼肝脏细胞质中存在特异性雌激素受体(E2的解离常数KD约为1.5×10^(-9) M)。(ii)雄性肝脏提供了“零”背景的理想实验对照,我们在雄性鳟鱼肝脏中,在E2刺激后,通过与Vg cDNA杂交追踪Vg mRNA的诱导动力学,以及(iii)使用一种原始的火箭免疫电泳技术检测血清中Vg的出现。雄性鳟鱼肝脏卵黄蛋白原模型以及我们开发的原始技术对于研究内源性和外源性因素对卵黄生成调节的不同步骤(受体、转录、翻译)的影响将非常有用。