Estrogen regulates the transcription of guppy isotocin receptors

Estrogen can regulate oxytocin receptor expression, which is mediated through estrogen receptors (ESRs) in mammals, initiating parturition. To further study the reproductive physiological process of ovoviviparous teleosts, guppies (Poecilia reticulata) were employed as the research model in the present study to identify the transcriptional regulation of ESRs on isotocin receptors (itrs). Since guppy embryos develop inside the ovary, in the present study, the levels of itrs in the ovarian stroma of pregnant female guppies treated with estradiol (E₂) in vitro were tested. E₂ increased only itr2 mRNA levels 3 h post-treatment, with no variation in itr1 mRNA expression levels. In vivo, pregnant guppies were immersed in different concentrations of E₂, significantly increasing the relative expression levels of itr1 and itr2 in the ovary. Moreover, based on dual-fluorescence in situ hybridization (ISH), both esrs and itrs mRNAs were localized in the same cells around the embryos in the ovary. To further investigate the regulation of itr transcription by estrogen, a luciferase reporter assay was performed, and the results demonstrated that E₂ treatment could induce E₂-dependent repression of luciferase activity in cells transfected with ESR1. However, overexpression of ESR2a or ESR2b caused a robust ligand-independent increase in itr2 promoter activity. Deletion analysis of the itr2 promoter indicated that there were novel potential ESR transcription factor-binding sites at -360 bp upstream of the 5' end of the itr2 promoter. Overall, our study provided novel results regarding the ESRs mediating the onset of parturition in ovoviviparous teleosts.