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岩鱼(许氏平鲉)中卵黄蛋白原(VTG)对4-壬基酚响应的蛋白质和基因表达

Protein and gene expression of VTG in response to 4-nonylphenol in rockfish (Sebastes schlegeli).

作者信息

Jung Jee-Hyun, Shim Won Joon, Addison R F, Baek Jae Min, Han Chang-Hee

机构信息

South Sea Institute, KORDI, 391 Jangbuk-ri Jangmok-myon, Geoje.

出版信息

Comp Biochem Physiol C Toxicol Pharmacol. 2006 Jun;143(2):162-70. doi: 10.1016/j.cbpc.2006.01.009. Epub 2006 Mar 10.

Abstract

To investigate the in vivo estrogenic activity of 4-nonylphenol (4-NP) on reproductive function, we have measured the expression level of rockfish vitellogenin (rVTG) mRNA, concentration of plasma VTG and levels of plasma estradiol-17beta(E(2)) and testosterone (T) by radioimmunoassay (RIA). An rVTG mRNA transcript of approximately 4.0 kb was extracted from hepatic tissue for Northern blot analysis, and the effects of 4-NP on rVTG mRNA expression in vivo with both male and female juvenile rockfish were examined. The level of rVTG mRNA expression was increased 48 h after injection with 4-NP of 10 mg/kg body mass and after injection in both male and female rockfish. The level of rVTG mRNA expression was increased 24 h after injection in male rockfish injected with 4-NP of 25 mg/kg body mass, while the level of rVTG mRNA expression was increased 12h after injection in female injected with same dosage. Concentrations of plasma rVTG in female injected with 4-NP of 10 and 25 mg/kg b.w. were increased 72 h after injection and reached 35 and 66 mg/mL, respectively. In male, rVTG concentrations with the dosage of 25 mg/kg body mass were increased 72 h after injection and reached to 14 and 65 mg/mL at 168 h, respectively. Plasma concentrations of T and E(2) in female and male injected with 4-NP were not significantly different between two 4-NP dosages. These results suggest that 4-NP may disrupt the reproductive system of immature rockfish by acting directly on vitellogenesis. Immature females injected with 4-NP were more sensitive than immature males, but VTG in immature males is likely to return to normal conditions slowly. There are obviously large interspecies differences in response of fish to 4-NP.

摘要

为研究4-壬基酚(4-NP)对生殖功能的体内雌激素活性,我们通过放射免疫分析(RIA)测定了岩鱼卵黄蛋白原(rVTG)mRNA的表达水平、血浆VTG浓度以及血浆雌二醇-17β(E₂)和睾酮(T)的水平。从肝脏组织中提取了约4.0 kb的rVTG mRNA转录本用于Northern印迹分析,并检测了4-NP对雌雄幼龄岩鱼体内rVTG mRNA表达的影响。注射10 mg/kg体重的4-NP后48小时,雌雄岩鱼体内rVTG mRNA表达水平均升高。注射25 mg/kg体重的4-NP后,雄性岩鱼注射后24小时rVTG mRNA表达水平升高,而雌性注射相同剂量后12小时rVTG mRNA表达水平升高。注射10和25 mg/kg体重的4-NP的雌性,注射后72小时血浆rVTG浓度升高,分别达到35和66 mg/mL。在雄性中,25 mg/kg体重剂量的rVTG浓度在注射后72小时升高,在168小时分别达到14和65 mg/mL。注射4-NP的雌雄岩鱼血浆中T和E₂的浓度在两种4-NP剂量之间无显著差异。这些结果表明,4-NP可能通过直接作用于卵黄生成来扰乱未成熟岩鱼的生殖系统。注射4-NP的未成熟雌性比未成熟雄性更敏感,但未成熟雄性中的VTG可能会缓慢恢复到正常水平。鱼类对4-NP的反应存在明显的种间差异。

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