Establishment of Reporter Yeasts for Guinea Pig and Syrian Hamster Aryl Hydrocarbon Receptor Ligand Activity
Establishment of Reporter Yeasts for Guinea Pig and Syrian Hamster Aryl Hydrocarbon Receptor Ligand Activity
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DOI：
10.3123/jemsge.28.167
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发表时间：
2006-11
影响因子：
1.7
通讯作者：
M. Kawanishi;Michiyo Sakamoto;Chiaki Shimohara;T. Yagi
中科院分区：
文献类型：
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作者：
M. Kawanishi;Michiyo Sakamoto;Chiaki Shimohara;T. Yagi
芳基碳氢化合物受体（AhR）的多态性可能会导致动物对芳基碳氢化合物易感性的遗传差异。 AhR 与芳基碳氢化合物结合后细胞内信号传导的激活与这些化学物质的毒性和致癌性高度相关。在这里，我们开发了两种共表达豚鼠和叙利亚仓鼠的 AhR 和 AhR 核易位蛋白 (Arnt) 蛋白的报告酵母，这两种蛋白分别被称为对 2,3,7,8-四氯二苯并-对二恶英 (TCDD) 最敏感和最具抵抗力的实验室啮齿动物。我们之前构建了表达人和小鼠 AhR/Arnt 的报告酵母。我们进行了报告基因检测来测量这些酵母中 TCDD、3-甲基胆蒽、β-萘黄酮和靛玉红的配体活性。配体处理诱导所有 4 种酵母菌株中报告质粒的 β-半乳糖苷酶活性呈剂量依赖性增加。测定表明，表达豚鼠 AhR/Arnt 的酵母对这些配体最敏感，而表达叙利亚仓鼠 AhR/Arnt 的酵母对这些配体最不敏感。表达人和小鼠 AhR/Arnt 的酵母介于两者之间。这些不同的配体活性反映了AhR/Arnt的物种特异性，并可能与啮齿动物对芳烃的敏感性有关。
Polymorphism of the aryl hydrocarbon receptor (AhR) presumably induces genetic difference in the susceptibility of animals to aryl hydrocarbons. The activation of intracellular signaling following AhR binding to aryl hydrocarbons is highly correlated with the toxicity and carcinogenicity of these chemicals. Here we developed two reporter yeasts coexpressing AhR and AhR nuclear translocator (Arnt) proteins of a guinea pig and a Syrian hamster, known as the most sensitive and most resistant laboratory rodents to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), respectively. We previously constructed reporter yeasts expressing human and mouse AhR/Arnt. We conducted reporter assays to measure ligand activities of TCDD, 3-methylcholanthrene, β-naphthoflavone and indirubin in these yeasts. Ligand treatment induced a dose-dependent increase in β-galactosidase activity from a reporter plasmid in all 4 yeast strains. The assays showed that yeast expressing guinea pig AhR/Arnt is most sensitive and yeast expressing Syrian hamster AhR/Arnt is most insensitive to these ligands. The yeasts expressing human and mouse AhR/Arnt were in-between. These different ligand activities reflect the species specificity of AhR/Arnt, and may be related to the susceptibility of rodents to aryl hydrocarbons.