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Intestinal cellular localization of PCNA protein and CYP1A mRNA in Atlantic salmon Salmo salar L. exposed to a model toxicant

机译:暴露于模型毒物的大西洋鲑鱼Salmo salar L.中PCNA蛋白和CYP1A mRNA的肠道细胞定位

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摘要

BackgroundThe aim of the study was to examine the intestinal cellular localization of proliferating cell nuclear antigen (PCNA) and cytochrome P450 A1 (CYP1A) expression in Atlantic salmon Salmo salar L. exposed to a model toxicant. The stress response was induced by intraperitoneal injection of four salmon with a single dose (50 mg/kg) of the CYP1A inducer β-naphthoflavone (BNF) and intestinal tissue (mid and distal intestine; MI and DI) was sampled seven days later. Samples for histology and gene transcription analysis were collected from four exposed fish and four control fish. PCNA was assessed by immunohistochemistry, CYP1A mRNA was studied by in situ hybridization (ISH) and finally the transcription of five genes was quantified by real-time quantitative RT-PCR (real-time RT-PCR); two detoxifying genes (CYP1A and glutathione S-transferase; GST), a stress marker gene (heat shock protein 70; HSP70), PCNA and a gene marker of apoptosis (caspase 6A).
机译:背景研究的目的是研究暴露于模型毒物的大西洋鲑鱼鲑的生长细胞核抗原(PCNA)和细胞色素P450 A1(CYP1A)表达在肠道中的细胞定位。腹腔内注射四只鲑鱼,用单剂量(50 mg / kg)CYP1A诱导剂β-萘黄酮(BNF)诱导肠应激(肠和中肠和远端肠; MI和DI),采集7天后取样。从四只裸露的鱼和四只对照鱼中收集用于组织学和基因转录分析的样品。免疫组化法检测PCNA,通过原位杂交(ISH)研究CYP1A mRNA的表达,最后通过实时定量RT-PCR(real-time RT-PCR)对5个基因的转录进行定量。两个解毒基因(CYP1A和谷胱甘肽S-转移酶; GST),应激标记基因(热激蛋白70; HSP70),PCNA和凋亡的基因标记(胱天蛋白酶6A)。

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