Generation of Knockout and Transgenic Zebrafish to Characterize Abcc4 Functions in Detoxification and Efflux of Lead
文献类型： 外文期刊
作者： Lu, Xing 1 ; Long, Yong 3 ; Li, Xixi 2 ; Zhang, Lang 2 ; Li, Qing 3 ; Wen, Hua 1 ; Zhong, Shan 2 ; Cui, Zongbin 3 ;
作者机构： 1.Chinese Acad Fishery Sci, Yangtze River Fisheries Res Inst, Key Lab Freshwater Biodivers Conservat, Minist Agr & Rural Affairs, Wuhan 430223, Peoples R China
2.Wuhan Univ, Dept Genet, Wuhan 430071, Peoples R China
3.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China
4.Guangdong Acad Sci, Guangdong Inst Microbiol, Guangdong Prov Key Lab Microbial Culture Collect, State Key Lab Appl Microbiol Southern China, Guangzhou 510070, Peoples R China
5.Hubei Prov Key Lab Allergy & Immunol, Wuhan 430071, Peoples R China
关键词： zebrafish; Abcc4; lead; detoxification; efflux transporter; transgenesis; knockout
期刊名称：INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES （ 影响因子：5.923； 五年影响因子：6.132 ）
ISSN：
年卷期： 2021 年 22 卷 4 期
页码：
收录情况： SCI
摘要： Lead (Pb) is one of the major heavy metals that are toxic to vertebrates and usually considered as environmental pollutants. ABCC4/MRP4 is an organic anion transporter that mediates cellular efflux of a wide range of exogenous and endogenous compounds such as cyclic nucleotides and anti-cancer drugs; however, it remains unclear whether ABCC4 and its orthologs function in the detoxification and excretion of toxic lead. In this study, we found that the transcriptional and translational expression of zebrafish abcc4 was significantly induced under lead exposure in developing zebrafish embryos and adult tissues. Overexpression of zebrafish Abcc4 markedly decreased the cytotoxicity and accumulation of lead in pig renal proximal tubule cell line (LLC-PK1 cells). To further understand the functions of zebrafish Abcc4 in lead detoxification, the clustered regularly interspaced palindromic repeats (CRISPR)/Cas9 system was used to create an abcc4(-/-) mutant zebrafish line. In comparison with the wild-type (WT) zebrafish, the abcc4(-/-) mutants showed a higher death rate and lead accumulation upon exposure to lead. Furthermore, a stable abcc4-transgenic zebrafish line was successfully generated, which exerted stronger ability to detoxify and excrete lead than WT zebrafish. These findings indicate that zebrafish Abcc4 plays a crucial role in lead detoxification and cellular efflux and could be used as a potential biomarker to monitor lead contamination in a water environment.
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