雌激素受体在间情期绵羊输卵管上皮细胞中的表达

Expression of estrogen receptors in ovine oviduct epithelium

为探讨膜性雌激素受体GPER(G protein-coupled estrogen receptor 1)及雌激素核受体ERα、ERβ在绵羊输卵管上皮细胞中的表达和定位,探讨三种受体在间情期绵羊输卵管中的表达规律,通过RT-qPCR方法和免疫印迹方法分别检测了三种受体基因mRNA水平和蛋白水平的表达差异;运用免疫组化法对输卵管组织中三种受体基因进行定位并对结果进行光密度值分析。结果,输卵管上皮细胞中GPER相对表达量高于ERα、ERβ(P<0.01),ERα的相对表达量高于ERβ(P<0.05)。免疫组化结果显示,GPER广泛分布于绵羊输卵管上皮细胞中,主要定位于细胞膜及细胞质。而ERα及ERβ主要分布于绵羊输卵管上皮细胞的细胞核。阳性细胞光密度分析结果与荧光定量RT-qPCR的分析结果相一致。上述结果表明,间情期绵羊输卵管上皮细胞中同时存在GPER、ERα、ERβ三种雌激素受体表达,且GPER的平均光密度、mRNA相对表达量最高,ERβ的平均光密度、mRNA相对表达量最低。

The objective of the experiment was to investigate the expression and localition of the estrogen receptors （GPER,ERα and ERβ） in ovine oviduct epithelium,and to explore the expression of the three receptors in the fallopian tube of sheep.Gene expression and protein levels was detected by reverse transcriptase quantitative real-time fluorescence polymerase chain reaction （RT-qPCR） and Western-blot,respectively.In result,the relative expression level of GPER mRNA was significantly higher than that of ERα and ERβ（P〈0.01）.The expression of ERα mRNA was higher than that of ERβ （P〈0.05）.The GPER receptor was widely distrbuted in the ovine oviduct epithelium,including cell-membrance and cytoplasm.Otherwise,the ERα and the ERβ were located on the nucleus.The analysis results of optical density in the immunopositive cells and RT-qPCR were in the similar trend.In conclusion,the three receptors were all existed in the ovine oviduct epithelium during the dioestrum cycle,and the average optical density and relative quantity of GPER were higher than that of the other two receptors.