Molecular cloning and mRNA expression of the vitellogenin and nuclear receptor gene induced by 17β-estradiol in the mud carp, Cirrhinus molitorella

Abstract

Vitellogenins (VTGs), the precursors of yolk proteins, are crucial for the embryonic development of teleosts, and have also been studied extensively as biomarkers for environmental estrogenic mimics. The cDNA of a VTG was isolated from the liver of male mud carp (Cirrhinus molitorella) following induction by 17β-estradiol (E(2)) using 3'- and 5'-RACE methods. It was 4,216 bp in length, and encoded a putative protein of 1,342 amino acids. This putative VTG contained complete portions of lipovitellin I (LVI) and phosvitin, but lacked the C-terminal half of LVII and was thus classed as a type I vitellogenin. In addition, the partial cDNA of estrogen receptors (ERα and ERβ) and androgen receptors (AR) were cloned. The mRNA expression of VTG, ERα, ERβ and AR were examined in the liver of juvenile mud carp exposed to E(2) (0, 5, 50 and 500 ng/l) in the water for 2, 4, 6 and 8 days. Quantitative reverse transcription PCR (RT-PCR) revealed a significant increase in transcript levels of VTG and ERα but not of ERβ and AR. VTG and ERα were expressed in a time-dose-dependent manner. Our results imply that ERα may be the principal subtype that regulates VTG, and that VTG mRNA of mud carp is an ideal biomarker to detect environmental estrogens.