Seok Eunju, Son Minyoung, Lee Seunghyung, Cheong Hee-Tae, Lee Sang-Hee
College of Animal Life Sciences, Kangwon National University, Chuncheon 24341, Republic of Korea.
College of Veterinary Sciences, Kangwon National University, Chuncheon 24341, Republic of Korea.
Animals (Basel). 2025 Jul 16;15(14):2100. doi: 10.3390/ani15142100.
Oocytes and cumulus cells undergo meiotic resumption and proliferation via gonadotropins and growth factors during maturation, and various small G proteins are activated when COCs undergo physiological changes. This study investigated the influence of gonadotropins and growth factors on Ras and its GTPases during porcine COC maturation. Unmatured COCs were treated with FSH, LH, or EGF for 44 h. The mRNA expression levels of the Ras subfamily (, , , and ), its GTPases ( and ), and proliferation factors (, , and ) were analyzed using RT-PCR. In contrast to other Ras subfamilies, R-Ras expression is upregulated during COC maturation. We evaluated the effects of FSH, LH, and EGF at various concentrations that most effectively regulated the expression of R-Ras and GTPases. The results demonstrated that 0.5 µg/mL FSH, 10 IU/mL human chorionic gonadotropin (hCG), and 10 ng/mL EGF effectively enhanced R-Ras expression and cell proliferation. FSH supplementation during porcine COC maturation significantly upregulated and expression, independent of LH and EGF, and downregulated expression. These results indicated that FSH regulates R-Ras expression, thereby promoting cell proliferation during COC maturation. These results provide fundamental knowledge for understanding the role of Ras and its family members in the development of follicular environments in pigs.
在成熟过程中,卵母细胞和卵丘细胞通过促性腺激素和生长因子进行减数分裂恢复和增殖,并且当卵丘卵母细胞复合体(COCs)发生生理变化时,各种小G蛋白被激活。本研究调查了促性腺激素和生长因子对猪COC成熟过程中Ras及其GTP酶的影响。未成熟的COCs用促卵泡激素(FSH)、促黄体生成素(LH)或表皮生长因子(EGF)处理44小时。使用逆转录聚合酶链反应(RT-PCR)分析Ras亚家族(R-Ras、H-Ras、K-Ras和N-Ras)、其GTP酶(Raf-1和MEK)以及增殖因子(c-Fos、c-Jun和Cyclin D2)的mRNA表达水平。与其他Ras亚家族不同,R-Ras在COC成熟过程中的表达上调。我们评估了最有效调节R-Ras和GTP酶表达的不同浓度的FSH、LH和EGF的作用。结果表明,0.5μg/mL FSH、10IU/mL人绒毛膜促性腺激素(hCG)和10ng/mL EGF有效增强了R-Ras表达和细胞增殖。在猪COC成熟过程中补充FSH显著上调了Raf-1和MEK的表达,独立于LH和EGF,并下调了Cyclin D2的表达。这些结果表明,FSH调节R-Ras表达,从而在COC成熟过程中促进细胞增殖。这些结果为理解Ras及其家族成员在猪卵泡环境发育中的作用提供了基础知识。