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摄食受油污污染的美北太平洋磷虾对大西洋鲱(Clupea harengus)的转录效应。

Transcriptional effects of dietary exposure of oil-contaminated Calanus finmarchicus in Atlantic herring (Clupea harengus).

机构信息

National Institute of Nutrition and Seafood Research (NIFES), Bergen, Norway.

出版信息

J Toxicol Environ Health A. 2011;74(7-9):508-28. doi: 10.1080/15287394.2011.550560.

Abstract

Suppression subtractive hybridization (SSH) cDNA library construction and characterization was used to identify differentially regulated transcripts from oil exposure in liver of male Atlantic herring (Clupea harengus) fed a diet containing 900 mg crude oil/kg for 2 mo. In total, 439 expressed sequence tags (EST) were sequenced, 223 from the forward subtracted library (enriched for genes putatively upregulated by oil exposure) and 216 from the reverse subtracted library (enriched for genes putatively downregulated by oil exposure). Follow-up reverse-transcription (RT) quantitative polymerase chain reaction (qPCR) analyses of gene transcription were conducted on additional herring exposed to food containing 9 (low), 90 (medium), and 900 (high) mg crude oil/kg feed for 2 mo. Chronic exposure of Atlantic herring to an oil-contaminated diet mediated upregulation of transcripts encoding antifreeze proteins, proteins in the classical complement pathway (innate immunity), and iron-metabolism proteins. Gene ontology (GO) analysis showed that "cellular response to stress," "regulation to biological quality," "response to abiotic stimuli," and "temperature homeostasis" were the most affected go at the biological processes level, and "carbohydrate binding," "water binding," and "ion binding" at the molecular function level. Of the genes examined with RT-qPCR, CYP1A, antifreeze protein, retinol binding protein 1, deleted in malignant brain tumor 1, and ovary-specific C1q-like factor demonstrated a significant upregulation. Myeloid protein 1, microfibrillar-associated protein 4, WAP65, and pentraxin were downregulated in liver of fish from the high exposure group. In conclusion, this study suggests that 2 mo of oil exposure affected genes encoding proteins involved in temperature homeostasis and possible membrane stability in addition to immune-responsive proteins in Atlantic herring.

摘要

采用抑制性消减杂交(SSH) cDNA 文库构建和鉴定技术,从摄食含 900mg 粗油/kg 饲料 2 个月的雄性大西洋鲱(Clupea harengus)肝脏中鉴定出与油暴露差异调节的转录本。总共测序了 439 个表达序列标签(EST),其中 223 个来自正向消减文库(富集了推测因油暴露而上调的基因),216 个来自反向消减文库(富集了推测因油暴露而下调的基因)。对另外一些摄食含 9(低)、90(中)和 900(高)mg 粗油/kg 饲料 2 个月的鲱鱼进行了基因转录的后续逆转录(RT)定量聚合酶链反应(qPCR)分析。大西洋鲱慢性暴露于受污染的油饮食中会导致编码抗冻蛋白、经典补体途径(先天免疫)蛋白和铁代谢蛋白的转录本上调。基因本体(GO)分析表明,在生物学过程水平上,“细胞对压力的反应”、“对生物质量的调节”、“对非生物刺激的反应”和“温度恒定性”是受影响最大的 GO,在分子功能水平上,“碳水化合物结合”、“水结合”和“离子结合”是受影响最大的 GO。在经 RT-qPCR 检验的基因中,CYP1A、抗冻蛋白、视黄醇结合蛋白 1、恶性脑肿瘤缺失蛋白 1 和卵巢特异性 C1q 样因子表现出显著上调。髓样蛋白 1、微纤维相关蛋白 4、WAP65 和 pentraxin 在高暴露组鱼的肝脏中下调。总之,本研究表明,2 个月的油暴露影响了编码与温度恒定性和可能的膜稳定性相关的蛋白质的基因,以及大西洋鲱的免疫反应蛋白。

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