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采用卵黄蛋白原同源放射免疫分析法,研究17β-雌二醇和鲤鱼促性腺激素对正常及垂体切除的欧洲雌性银鳗(Anguilla anguilla L.)卵黄生成的影响。

Effects of 17 beta-estradiol and carp gonadotropin on vitellogenesis in normal and hypophysectomized European silver female eel (Anguilla anguilla L.) employing a homologous radioimmunoassay for vitellogenin.

作者信息

Burzawa-Gerard E, Dumas-Vidal A

机构信息

Laboratoire de Physiologie Générale et Comparée MNHN, Paris, France.

出版信息

Gen Comp Endocrinol. 1991 Nov;84(2):264-76. doi: 10.1016/0016-6480(91)90049-c.

Abstract

The European silver eels are at the early stages of vitellogenesis before the marine reproductive migration. Vitellogenesis was induced by 17 beta-estradiol (E2) alone and by a purified carp gonadotropin (cGTH). We studied and compared their effects on plasma vitellogenin (Vg) levels and ovarian yolk contents in female normal (N) and hypophysectomized (H) eels for both treatments. To this purpose an homologous radioimmunoassay (RIA) was established. Eel Vg was purified to homogeneity on 0.1% SDS-Electrophoresis. Native Vg has a molecular weight of 340 +/- 15 kilodalton (kDa) and was partially separated into subunits. The RIA was established with a sensitivity of 1.1 ng and was specific for eel Vg. In control (N and H) silver eels, plasma Vg levels were 0.04 +/- 0.02 microgram/ml and unchanged throughout the experiment. Similarly, yolk was indetectable in control ovarian extracts. E2 treatment increased plasma Vg levels proportionally with time to 783.4 +/- 130.7 micrograms/ml in N eels. The same profile was seen in H eels but terminally the mean value was 36.7 times lower than in N eels (P less than 0.01). Yolk at 0.005 microgram/g in N eels was indetectable in H eels. cGTH treatment gave a biphasic kinetic change: plasma Vg increased within 12 days, peaked at 93.6 +/- 13.0 micrograms/ml at 20 to 24 days, and stabilized to decrease at 40.2 +/- 7.5 micrograms/ml. The gonadosomatic index (GSI) increased alongside the yolk content (980.4 +/- 153.1 micrograms Vg/g). The kinetic profile for H eels was different: a peak was not apparent, rather there was a delayed increase, and at 67 days levels were still 8.23 times lower than in N eels (P less than 0.01). The GSI increased as the yolk content to 202.7 +/- 64.8 micrograms Vg/g ovary showing an ovarian incorporation of Vg in H eels.

摘要

欧洲银鳗在进行海洋繁殖洄游之前正处于卵黄发生的早期阶段。单独使用17β-雌二醇(E2)和纯化的鲤鱼促性腺激素(cGTH)均可诱导卵黄发生。我们研究并比较了这两种处理对正常(N)和垂体切除(H)雌性鳗鱼血浆卵黄蛋白原(Vg)水平和卵巢卵黄含量的影响。为此建立了一种同源放射免疫分析(RIA)方法。鳗鱼Vg在0.1% SDS-聚丙烯酰胺凝胶电泳上被纯化至均一。天然Vg的分子量为340±15千道尔顿(kDa),并部分分离成亚基。所建立的RIA灵敏度为1.1 ng,对鳗鱼Vg具有特异性。在对照(N和H)银鳗中,血浆Vg水平为0.04±0.02微克/毫升,在整个实验过程中保持不变。同样,在对照卵巢提取物中未检测到卵黄。E2处理使N鳗鱼的血浆Vg水平随时间成比例增加至783.4±130.7微克/毫升。H鳗鱼也呈现相同的变化趋势,但最终平均值比N鳗鱼低36.7倍(P<0.01)。N鳗鱼中卵黄含量为0.005微克/克,而H鳗鱼中未检测到。cGTH处理呈现双相动力学变化:血浆Vg在12天内增加,在20至24天达到峰值93.6±13.0微克/毫升,然后稳定下降至40.2±7.5微克/毫升。性腺指数(GSI)随卵黄含量增加(980.4±153.1微克Vg/克)。H鳗鱼的动力学变化不同:未出现明显峰值,而是延迟增加,在67天时水平仍比N鳗鱼低8.23倍(P<0.01)。GSI随卵黄含量增加至202.7±64.8微克Vg/克卵巢,表明H鳗鱼卵巢中有Vg掺入。

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