Investigation of the interaction between bta-miR-222 and the estrogen receptor alpha gene in the bovine ovarium.

The aim of the present study was to investigate the posttranscriptional regulation of bta-miR-222 and the estrogen receptor (ER)-alpha gene in cows. The preovulatory follicles (POFs) were detected and ER-alpha levels in the corpus luteum (CL) were measured via immunofluorescent method. Afterwards, an ELISA test was performed to detect estradiol- and progesterone-active follicles, and the expression levels of ER-alpha, progesterone receptor (PR), and bta-miR-222 were measured via qRT-PCR. Finally, a western blot analysis was performed to determine ER-alpha protein levels. Immunofluorescence staining was highly positive in the follicular phase, showing ER-alpha and PR immunopositivity. In the luteal phase, ER-alpha immunopositivity decreased in lutein cells, whereas the PRs were observed to be similar in intensity to those in follicular phase. While the estradiol levels were higher in the POFs, the progesterone level was higher in the CL. In the CL, the transcription levels of ER-alpha and PR mRNA were observed to be the same as in the POFs; however, the expression of bta-miR-222 was lower. In the CL, the transcription level of ER-alpha mRNA was lower than that of PR mRNA, and the expression level of bta-miR-222 was higher than that of PR mRNA. The results of the western blot analysis show that the ER-alpha protein level in the CL was lower than that in the POFs. Taken together, our results here suggest that there is a negative correlation between bta-miR-222 and ER-alpha expression during follicular development in cow ovaries.

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