title: Investigation of effect of 17α-ethinylestradiol on vigilin expression using an isolated recombinant antibody.
abstract: Vigilin, a highly conserved protein from yeast to mammals, is a multifunctional protein in eukaryotic organisms. One biological function of vigilin is to stabilize the expression level of vitellogenin (VTG). This study aimed to develop vigilin as a new estrogen-inducible biomarker that correlates with the widely applied estrogen-inducible biomarker VTG and expand the ability to detect it in various species. Here, a recombinant monoclonal antibody with high specificity against the conserved C-terminal region of vigilin from zebrafish (Danio rerio) was successfully isolated from a phage display antibody library and found to recognize vigilin proteins from multiple vertebrate species. The effect of 17α-ethinylestradiol (EE2) on vigilin expression in the liver of zebrafish and juvenile crucian carp (Carassius auratus) was investigated. Although vigilin mRNA was expressed in all tissues analyzed from male zebrafish, vigilin protein was detected exclusively in the testis of male zebrafish, as well as the liver of female zebrafish and juvenile crucian carp at a lower level without exposure to EE2. Significant induction of vigilin mRNA by exposure to EE2 was observed in the liver and testis of male zebrafish, even at a low dose of 6.25 ng/L (21.09 pmol/L). In Hela cells, the expression of vigilin coincided with high protein synthesis activity but not dose-dependently by EE2 exposure. Therefore, the recombinant antibody may be used as a detection tool to screen for mammalian cell lines or organs with estrogen-inducible expression of vigilin.
authors.affiliation: State Key Laboratory of Fresh water Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, 7 Southern East Lake Road, Wuhan 430072, PR China.
authors.forename: Liqiao
authors.initials: L
authors.lastname: Zhong
authors.affiliation: State Key Laboratory of Fresh water Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, 7 Southern East Lake Road, Wuhan 430072, PR China.
authors.forename: Li
authors.initials: L
authors.lastname: Yuan
authors.affiliation: State Key Laboratory of Fresh water Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, 7 Southern East Lake Road, Wuhan 430072, PR China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Rao
authors.affiliation: State Key Laboratory of Fresh water Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, 7 Southern East Lake Road, Wuhan 430072, PR China; University of Chinese Academy of Sciences, Yuquan Road 19A, Beijing 100039, PR China.
authors.forename: Zhouquan
authors.initials: Z
authors.lastname: Li
authors.affiliation: State Key Laboratory of Fresh water Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, 7 Southern East Lake Road, Wuhan 430072, PR China; University of Chinese Academy of Sciences, Yuquan Road 19A, Beijing 100039, PR China.
authors.forename: Qilin
authors.initials: Q
authors.lastname: Gu
authors.affiliation: State Key Laboratory of Fresh water Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, 7 Southern East Lake Road, Wuhan 430072, PR China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Long
authors.affiliation: State Key Laboratory of Fresh water Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, 7 Southern East Lake Road, Wuhan 430072, PR China. Electronic address: zhxh@ihb.ac.cn.
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Fresh water Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, 7 Southern East Lake Road, Wuhan 430072, PR China.
authors.forename: Zongbin
authors.initials: Z
authors.lastname: Cui
authors.affiliation: State Key Laboratory of Fresh water Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, 7 Southern East Lake Road, Wuhan 430072, PR China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Fresh water Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, 7 Southern East Lake Road, Wuhan 430072, PR China. Electronic address: hpdai@ihb.ac.cn.
authors.forename: Heping
authors.initials: H
authors.lastname: Dai
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D002347:Carps / Q000235:genetics / Q000502:physiology; D004997:Ethinyl Estradiol / Q000633:toxicity*; D005260:Female; D005786:Gene Expression Regulation / Q000187:drug effects*; D006367:HeLa Cells; D006801:Humans; D008099:Liver / Q000187:drug effects; D008297:Male; D011485:Protein Binding; D012333:RNA, Messenger / Q000235:genetics; D016601:RNA-Binding Proteins / Q000032:analysis / Q000235:genetics*; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification; D013737:Testis / Q000187:drug effects; D014819:Vitellogenins / Q000378:metabolism; D014874:Water Pollutants, Chemical / Q000633:toxicity; D015027:Zebrafish / Q000235:genetics / Q000502:physiology
chemical_list: D000911:Antibodies, Monoclonal; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D011994:Recombinant Proteins; D014819:Vitellogenins; D014874:Water Pollutants, Chemical; C075197:high density lipoprotein binding protein; D004997:Ethinyl Estradiol
keywords: 17α-ethinylestradiol (EE2); Crucian carp; Single-chain variable fragment (scFv); Vigilin; Zebrafish; phage display