Mfn2在不同繁殖生理时期牦牛卵巢和输卵管中的表达及定位

Distribution and Expression of Mfn2 in Ovary and Fallopian Tube of Yaks at Different Reproductive Physiological Periods

为探索线粒体融合蛋白2(Mfn2)在不同繁殖生理时期牦牛卵巢和输卵管中的表达情况与分布特征,选取处于不同繁殖生理时期健康牦牛15头,分为3组,每组5头,屠宰后采集处于卵泡期、黄体期和妊娠期牦牛的卵巢和输卵管组织样品,分为6个组,分别提取各组卵巢和输卵管组织样品总RNA和蛋白,用实时荧光定量PCR(RT-qPCR)和Western blot检测Mfn2 mRNA及其蛋白在6个组的组织中的表达情况;用IHC法检测Mfn2蛋白在组织中的定位。RT-qPCR检测结果表明,6个组的组织中Mfn2 mRNA均有表达且存在差异,Mfn2 mRNA在妊娠期表达量最高,其次为黄体期,卵泡期最低,且各组之间差异显著(P<0.05)。Western blot结果发现,Mfn2蛋白在6个组的组织中均有所表达。在卵巢组织中,妊娠期最高,与黄体期和卵泡期差异显著(P<0.05),黄体期与卵泡期差异显著(P<0.05)。而在输卵管中,黄体期最高,与妊娠期差异显著(P<0.05);在卵泡期次之,与妊娠期差异显著(P<0.05),与黄体期差异不显著(P>0.05)。IHC结果显示,在卵巢中,Mfn2主要分布在卵巢卵泡膜、黄体细胞、生殖上皮,在输卵管中主要分布在黏膜上皮和肌层中,说明Mfn2在不同繁殖生理时期牦牛的卵巢和输卵管中均有所表达且有差异。

To explore the expression and distribution of Mfn2 in yak ovaries and fallopian tubes during different reproductive physiological periods, 15 yaks were selected and divided into 3 groups.Ovaries and fallopian tubes were collected from yaks in the luteal phase, follicular phase, and pregnancy phase and divided into 6 groups;total RNA and protein were extracted from each group.RT-qPCR and Western blot techniques were used to detect the relative expressions of Mfn2 mRNA and protein;while IHC was used to analyze distribution of Mfn2 protein in each tissue.The results showed that Mfn2 mRNA expressed in all 6 groups.In the ovaries and fallopian tubes, relative expression of Mfn2 mRNA during pregnancy phase was highest(P<0.05),followed by the luteal phase(P<0.05) and lowest in the follicular phase(P<0.05);Mfn2 protein also expressed in all 6 groups.In the ovaries, it was highest in the pregnancy phase and differed significantly from the luteal phase and follicular phase(P<0.05).However in the fallopian tubes, it was highest in the luteal phase, with significantly difference from the pregnancy phase(P<0.05),followed by the follicular phase, which was not significantly from the luteal phase(P>0.05).In addition, Mfn2 protein distribution in the ovaries was mainly observed in theca follicle, corpus luteum verum, and germinal cells;in the fallopian tubes, Mfn2 expressed in mucosal epithelium and muscularis.The results showed that Mfn2 was differentially expressed in the ovaries and fallopian tubes of yaks at different reproductive physiological periods.