Transcriptome comparisons detect new genes associated with apoptosis of cattle and buffaloes preantral follicles

BACKGROUND: To develop new breeding technology to improve the breeding ability of bovine, it is the development trend to find the main reason for the occurrence of atresia in these organisms. Transcriptomes of small (100-120 μm) and large (200-220 μm) preantral follicles from cattle and buffalo ovaries were evaluated in vivo and in vitro to understand the transcriptional modulation in preantral follicles that leads to the phenomenon of atresia.

METHODS: The preantral follicles were checked as dead, damage, or live follicles in vivo and in vitro by using trypan blue then bisbenzimide and propidium iodine. Transcriptomes of small (100-120 μm) and large (200-220 μm) preantral follicles of cattle and buffalo were evaluated in vivo and in vitro by microarray and RT-PCR. Healthy preantral follicles were selected based on staining results, and then RNA was extracted from them.

RESULTS: The viability percentage of preantral follicles in cattle was higher (26.7% and 20%) than buffalo (10%) in vivo and in vitro, respectively. According to the microarray data analysis for cattle preantral follicles, only eleven genes were detected corresponding to five upregulated and six downregulated in large size (200-220 μm) compared to small (100-120 μm) size preantral follicles, while in buffalo, 171 genes were detected (92 upregulated and 79 downregulated) in large size compared to small preantral follicle size. The results of RT-PCR of the selected genes (FASTKD1, BAG2, RHOB, AGTR2, MEF2C, BCL10, G2E3, TM2D1, IGF-I, IGFBP3, PRDX3, and TRIAP1) validated the microarray results. In conclusion, the data of gene expression showed significant differences between small and large sizes in both buffalo and cattle preantral follicles.

CONCLUSION: Apoptotic genes were upregulated in the large preantral follicle compared with the small preantral follicles. Moreover, the expression level of these apoptotic genes was significantly upregulated in buffalo than in the cattle. Most of these genes were significantly upregulated in the large buffalo preantral follicle compared with the small size. However, anti-apoptotic genes were upregulated in large cattle preantral follicle and downregulated in large buffalo preantral follicle.