浑河流域野生鲫鱼卵黄蛋白原基因表达

Vitellogenin mRNA Expression in Wild Crucian Carp(Carassius auratus auratus)from Hun River

为揭示浑河流域水环境内分泌干扰物对水生态的潜在风险,利用兼并引物扩增获得鲫鱼卵黄蛋白原(Vtg)和核糖体蛋白L-7(RPL-7)基因部分碱基序列(分别为825和450bp),建立以RPL-7为内参基因、定量鲫鱼Vtg基因表达的实时定量逆转录聚合酶链反应(qRT-PCR)方法,并将该方法用于定量浑河野生鲫鱼肝组织Vtg基因表达分析。结果显示:与上游对照点(S1)相比,7月下游各点(S4～S8)雄鱼、S4和S6点雌鱼肝组织的VtgmRNA表达水平皆显著升高(P<0.05);11月在雄鱼中未检出VtgmRNA的有效表达,雌鱼也仅在S2和S3点的表达水平升高(P<0.05)。研究表明,浑河流域野生鲫鱼尤其是在7月明显受到了环境雌激素类物质的影响。另外,qRT-PCR方法能够灵敏检测出鲫鱼Vtg基因表达的时空差异。

To evaluate the potential ecological risk caused by endocrine disrupting chemicals in Hun River,a partial vitellogenin（Vtg） and ribosomal protein L7（RPL-7） genes in crucian carp（Carassius auratus auratus） were isolated,amplified and sequenced（825 bp and 450 bp）.According to the two gene sequences,a real-time quantitative reverse-transcription polymerase chain reaction（qRT-PCR） assay was developed for quantifying the Vtg mRNA expression in crucian carp,using RPL-7 as a reference gene.The established qRT-PCR method was applied to quantify Vtg mRNA expression in livers of wild crucian carp from Hun River.Results showed that the Vtg mRNA expressions in males from S4 to S8（down-stream sites）increased to be significantly higher than those from S1（up-stream reference site） in July（P0.05）.Similar results were also found in female crucian carp of S4 and S6 in July（P0.05）.In November,no significant Vtg mRNA expression was detected in males.Increased Vtg mRNA expression was only observed in females of S2 and S3（down-stream sites）（P0.05）.The study suggested that the wild crucian carp in Hun River were affected by the environmental estrogens especially in July.The present qRT-PCR method can be applied to detect the spatial and temporal differences of Vtg mRNA expressions in crucian carp.