miR-375 mediates CRH signaling pathway in inhibiting E2 synthesis in porcine ovary

Abstract

The corticotropin-releasing hormone (CRH) signaling system is involved in numbers of stress-related physiological and pathological responses，including its inhibiting effects on estradiol (E2) synthesis and follicular development in the ovary. In addition, there are reports that microRNAs (miRNAs) can control the function of animal reproductive system. The aim of present study was to investigate the functions of miR-375 and the relationship between miR-375 and CRH signaling molecules in the porcine ovary. First, our common PCR results show that miR-375 and the CRH receptor 1 (CRHR1) are expressed in porcine ovary, whereas CRH receptor 2 (CRHR2) is not detected. We further have located the cell types of miR-375 and CRHR1 by in situ hybridization (ISH), and the results show that miR-375 is located only in the granulosa cells, whereas CRHR1 is positive in all of granulosa cells and oocytes, inferring that miR-375 and CRHR1 are co-localized in granulosa cells. Second, we show that overexpression of miR-375 in cultured granulosa cells suppresses the E2 production, while miR-375 knockdown demonstrates the opposite result. Besides, our in vitro results demonstrate that miR-375 mediates the signaling pathway of CRH inhibiting E2 synthesis. Finally, our data show that the action of miR-375 is accomplished by directly binding to the 3'UTR of specificity protein1 (SP1) mRNA to decrease the SP1 protein level. Thus, we conclude that miR-375 is a key factor in regulating E2 synthesis by mediating the CRH signaling pathway.

Gene Expression Profiling in Ovaries and Association Analyses Reveal HEP21 as a Candidate Gene for Sexual Maturity in Chickens

“…A total of 619 differentially expressed genes were enriched in signaling receptor activity. Of these genes, corticotropin releasing hormone receptor 1 (CRHR1), platelet activating factor receptor (PTAFR), and 5-hydroxytryptamine receptor 4 (HTR4) are all highly expressed in ovaries, are involved in proliferation and apoptosis of ovarian cells, and play important roles in the development of the ovaries [27][28][29][30].…”

Effects of melatonin on the synthesis of estradiol and gene expression in pig granulosa cells

“…The cells were cultured in 5% CO 2 at 37°C. Porcine GCs were cultured in 10% fetal calf serum‐supplemented DMEM/F12 for 72 hours before further treatment, and the culture medium was refreshed every 24 hours …”

NRDR inhibits estradiol synthesis and is associated with changes in reproductive traits in pigs

“…Experiments were performed six times. P4 and E2 were analyzed using radioimmunoassay (RIA) reagents provided by the Beijing North Institute Biological Technology (Beijing, China), which have been validated for use in cell culture media (Liu et al, ; Yu et al, ), according to the manufacturer’s recommendations. The minimum detectable concentrations were 2 pg/ml for E2 and 0.2 ng/ml for P4.…”