17β—雌二醇对雄性金鱼卵黄原蛋白的诱导作用

采用腹腔注射17β-雌二醇的方法诱导雄性金鱼卵黄原蛋白产生,注射浓度为0.05mg·g-1BW,诱导2周后取尾静脉血,离心分离血浆,进行血浆常规聚丙烯酰胺凝胶电泳,通过对卵黄原蛋白特性基团磷、脂和糖蛋白的染色,确定了卵黄原蛋白在电泳图谱上的位置,开发了一种简便、高效的定性卵黄原蛋白的聚丙烯酰胺凝胶电泳法。电泳结果表明,在0.05mg·g-1BW的注射浓度下,2周后17β-雌二醇诱导了雄性金鱼卵黄原蛋白产生,并通过ELISA检测卵黄原蛋白的平均含量为690.2ng·mL-1,与对照组雄性金鱼平均含量为10.7ng·mL-1的差异极显著(P<0.01),比雌性对照组检出量285.5ng·mL-1高1倍多;17β-雌二醇诱导组雄鱼血浆钙离子和血总蛋白含量明显增加。

The identification of vitellogenin induction by 17β-estradiol in male Carassius auratus

The vitellogenin of male goldf ish ( Carassius auratus ) was induced by intraperitoneal injection of 0. 05 mg..g - 1 BW of 17....estradiol dissolved in ethanol. After two weeks induction, the blood was collected from caudal vessel, then centrifuged at 8 000 r..min - 1 for 5 min. After plasma native..PAGE, the gel was stained with the method of CBB G250.., phospho.., lipo.. and glyco..staining respectively. the location of vitellogenin in the gel was identified. This method provides a practical and ef fective way of vitellogenin identif ication. The result of native.. PAGE showed that, after the injection of 0. 05 mg ..g - 1BW of 17....estradiol for two weeks, the male fish can synthesize vitellogenin. The vitellogenin average content in E2 group was 690. 2 ng ..mL - 1 by ELISA, it was signif icant ly different from male control group( P < 0. 01) , whose average content was 10. 7 ng..mL - 1 , and it was also much higher than that of the female control group, who se average content was 285. 5 ng..mL - 1 . Compared w ith control male group, the Ca2 and plasma total protein content of the E2 exposed male group was increased signif icant ly. In control group, the Ca2 contents in plasma of male and female control groups were 1. 6 mmol..L- 1 and 1. 7 mmol..L- 1, respectively. After two..weeks treatment of E2, Ca2 content of male groups was 50% higher than that of male contro l group, which was 2. 3 mmol..L - 1 . The contents of total protein in the control male and female groups were 3253. 1 mg..L- 1 and 7071. 0 mg..L- 1, respectively. After two..weeks treatment of E2, the total protein content in plasma was 9616. 6 mg..L- 1, which is 3..folder higher than that of the control male group. So E2 not only can induce male goldf ish to synthesize vitellogenin but also can alter the physical index of f ish plasma.