Abstract
Compared with other livestock, fertility is low in jennies. Although the majority of jennies shows normal ovulation throughout the year, some jennies display hemorrhagic anovulatory follicles (HAF) in the middle of the winter or summer. Based on our previous results (https://doi.org/10.1111/rda.13883), this research was carried out in from November to January of the next year. In experiment 1, a total of 24 Yangyuan jennies (4∼8 years old) were included, with half of them showing normal preovulatory follicles (control group, Con-G) while the other half had HAF ad diagnosed by transrectal ultrasound (HAF-G). The content of the preovulatory follicle or HAF was collected from each jenny by ultrasound-guided ovum pick-up. The follicular fluid (FF) and granulosa cells (GCs) were used for omics analysis. The results of metabolomics analysis showed that the level of arachidonic acid (AA) was significantly increased in HAF-G (P < 0.05). According to transcriptomics results, the mRNA levels of arachidonate 5-lipoxygenase gene (ALOX5) was significantly increased (P < 0.05) while the level of cytochrome P450 family 2 subfamily J member 2 gene (CYP2J2) was significantly decreased in HAF-G (P < 0.05). However, there was no significant difference between the two groups in the expressionsof prostaglandin-endoperoxide synthase 1 gene (COX1) and COX2 (P > 0.05). Meanwhile, the expressions of FSHR, LHCGR and CYP19A1 were significantly decreased in the HAF-G compared with the Con-G (P < 0.05). In order to pursue the molecular mechanism of AA regulating the functions of granulosa cell, the experiment two was carried out. The mice GCs from antral follicles were cultured in media supplemented with 0, 50 μM or 100 μM AA respectively, and both of cell viability and some genes’ expressions were assayed. The results demonstrate that the survival of GCs and mRNA expressions of FSHR and LHCGR were higher in 50 μM AA group than the control group (P < 0.05). However, when the AA concentration was increased to 100 μM in the medium, the survival rates and expression of FSHR and LHCGR were decreased in comparison to the control group (P < 0.05). Moreover, 100 μM AA increased the expression of ALOX5 and decreased the expression of CYP2J2 compared with 50 μM AA, which was similar to GCs from HAF of jennies. In conclusion, the occurrence of HAF in jennies may be related to the accumulation of AA in follicular fluid of the preovulatory follicles which caused abnormal expression of FSHR, LHCGR and ALOX5 on GCs leading to anovulation and HAF.
Published Version
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