Control of transcriptional repression of the vitellogenin receptor gene in largemouth bass (Micropterus salmoides) by select estrogen receptors isotypes

• Published in: Toxicological sciences Vol. 141; no. 2; pp. 423 - 431
• Main Authors: Dominguez, Gustavo A, Bisesi, Jr, Joseph H, Kroll, Kevin J, Denslow, Nancy D, Sabo-Attwood, Tara
• Format: Journal Article
• Language: English
• Published: United States Oxford University Press 01.10.2014
• Subjects: Animals; Base Sequence; Bass - genetics; Bass - metabolism; Benzhydryl Compounds - pharmacology; Binding Sites; Cloning, Molecular; Down-Regulation; Egg Proteins - drug effects; Egg Proteins - genetics; Egg Proteins - metabolism; Estradiol - pharmacology; Estrogen Receptor alpha - agonists; Estrogen Receptor alpha - genetics; Estrogen Receptor alpha - metabolism; Estrogen Receptor beta - agonists; Estrogen Receptor beta - genetics; Estrogen Receptor beta - metabolism; Ethinyl Estradiol - pharmacology; Fish Proteins - drug effects; Fish Proteins - genetics; Fish Proteins - metabolism; Genes, Reporter; HEK293 Cells; Humans; Insulin - pharmacology; Molecular Sequence Data; Phenols - pharmacology; Promoter Regions, Genetic; Receptors, Cell Surface - drug effects; Receptors, Cell Surface - genetics; Receptors, Cell Surface - metabolism; RNA, Messenger - metabolism; Sp1 Transcription Factor - metabolism; Transcription, Genetic - drug effects; Transfection; estrogen; insulin; vitellogenin receptor; ethinylestradiol; estrogen receptors; largemouth bass; bisphenol-A
• ISSN: 1096-6080; 1096-0929
• DOI: 10.1093/toxsci/kfu145

The vitellogenin receptor (Vtgr) plays an important role in fish reproduction. This receptor functions to incorporate vitellogenin (Vtg), a macromolecule synthesized and released from the liver in the bloodstream, into oocytes where it is processed into yolk. Although studies have focused on the functional role of Vtgr in fish, the mechanistic control of this gene is still unexplored. Here we report the identification and analysis of the first piscine 5' regulatory region of the vtgr gene which was cloned from largemouth bass (Micropterus salmoides). Using this putative promoter sequence, we investigated a role for hormones, including insulin and 17β-estradiol (E2), in transcriptional regulation through cell-based reporter assays. No effect of insulin was observed, however, E2 was able to repress transcriptional activity of the vtgr promoter through select estrogen receptor subtypes, Esr1 and Esr2a but not Esr2b. Electrophoretic mobility shift assay demonstrated that Esr1 likely interacts with the vtgr promoter region through half ERE and/or SP1 sites, in part. Finally we also show that ethinylestradiol (EE2), but not bisphenol-A (BPA), represses promoter activity similarly to E2. These results reveal for the first time that the Esr1 isoform may play an inhibitory role in the expression of LMB vtgr mRNA under the influence of E2, and potent estrogens such as EE2. In addition, this new evidence suggests that vtgr may be a target of select endocrine disrupting compounds through environmental exposures.