ZFIN ID: ZDB-PUB-140813-7
Investigation of effect of 17α-ethinylestradiol on vigilin expression using an isolated recombinant antibody
Zhong, L., Yuan, L., Rao, Y., Li, Z., Gu, Q., Long, Y., Zhang, X., Cui, Z., Xu, Y., Dai, H.
Date: 2014
Source: Aquatic toxicology (Amsterdam, Netherlands) 156: 1-9 (Journal)
Registered Authors: Cui, Zongbin, Gu, Qilin, Long, Yong
Keywords: 17α-ethinylestradiol (EE2), Crucian carp, Single-chain variable fragment (scFv), Vigilin, Zebrafish, phage display
MeSH Terms: Animals; Antibodies, Monoclonal/metabolism*; Carps/genetics; Carps/physiology; Ethinyl Estradiol/toxicity* (all 22) expand
PubMed: 25112681 Full text @ Aquat. Toxicol.
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 6.25ng/L (21.09pmol/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.
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