不同繁殖期牦牛主要生殖器官中Uhrf1的表达与定位分析

Expression and localization of Uhrf1 in the major reproductive organs of yaks during different reproductive cycles

摘要:

Uhrf1是一种有多结构域和功能的表观遗传因子，在DNA甲基化、细胞代谢及细胞增殖中发挥关键作用。为探究Uhrf1在母牦牛生殖生理过程中的作用，本研究采集3种繁殖期(卵泡期、黄体期和妊娠期)健康牦牛的3种生殖器官(卵巢、输卵管和子宫)，共9组，采用实时荧光定量PCR (real-time fluorescence quantitative PCR,RT-qPCR)、蛋白免疫印迹技术(Western blotting)和免疫组织化学(immunohistochemistry,IHC)法检测Uhrf1的表达和定位情况。RT-qPCR和Western blotting结果显示，Uhrf1在卵泡期输卵管中的表达量最高，妊娠期子宫次之，黄体期子宫最低(P<0.05)。IHC结果显示，Uhrf1主要在卵巢生殖上皮、卵泡膜、卵泡颗粒层、黄体细胞、输卵管黏膜上皮细胞、子宫腺(uterine glands,UG)中表达。本研究表明Uhrf1在不同繁殖期母牦牛主要生殖器官中的表达存在差异，提示其在牦牛生殖生理过程中的调控作用具有重要意义。

Abstract:

Uhrf1 is a multi-domain and multifunctional epigenetic regulator playing key roles in DNA methylation, cell metabolism, and cell proliferation. To investigate the role of Uhrf1 in the reproductive physiology of female yaks, we collected three reproductive organs (ovaries, oviducts, and uteri) from healthy yaks during three reproductive phases (follicular, luteal, and gestational phases), with a total of nine groups. Real-time fluorescence quantitative PCR (RT-qPCR), Western blotting, and immunohistochemistry (IHC) were employed to determine the expression levels of Uhrf1 and the subcellular localization of this protein. RT-qPCR and Western blotting results showed that Uhrf1 was expressed highest in the oviduct during the follicular phase, moderate expression in the uterus during the gestational phase, and the lowest expression in the uterus during the luteal phase (P<0.05). IHC results showed that Uhrf1 was mainly expressed in the ovarian germinal epithelium, theca follicular, follicular granulosa, luteal cells, oviduct mucosal epithelial cells, and uterine glands (UG) of yaks. In conclusion, Uhrf1 was differentially expressed in the major reproductive organs during the reproductive cycle of female yaks, indicating its important regulatory role in the reproductive physiology of yaks.