| 摘要: |
| 【目的】探究透明质酸合成酶2（hyaluronate synthase 2，HAS2）和前列腺素内过氧化物合酶2（prostaglandin endoperoxide synthase 2，PTGS2）在牦牛不同时期卵巢中的定位与表达情况，为进一步提高耗牛生育能力提供理论依据。【方法】采集临床健康牦牛不同时期（卵泡期、黄体期、妊娠期）的卵巢作为试验样品，采用实时荧光定量PCR（qRT-PCR）、Western blotting方法检测牦牛不同时期卵巢中HAS2、PTGS2基因水平和蛋白水平的表达量，采用免疫组化（IHC）法检测HAS2和PTGS2在牦牛不同时期卵巢中的分布情况。【结果】HAS2基因在耗牛黄体期卵巢的表达量显著高于卵泡期和妊娠期（P＜0.05），在卵泡期卵巢的表达量显著高于妊娠期（P＜0.05）；PTGS2基因在黄体期卵巢的表达量显著高于其他2个时期（P＜0.05），在卵泡期卵巢的表达量显著高于妊娠期（P＜0.05）。HAS2蛋白在耗牛3个时期卵巢中均有表达，其中在黄体期卵巢的表达量显著高于其他2个时期（P＜0.05），且妊娠期卵巢的表达量显著高于卵泡期（P＜0.05）；PTGS2蛋白在黄体期卵巢的表达量显著高于其他2个时期（P＜0.05），且卵泡期卵巢的表达量显著高于妊娠期（P＜0.05）。IHC试验表明，HAS2和PTGS2蛋白在耗牛卵巢黄体细胞中表达量最高，在卵泡颗粒层和卵丘细胞中也有表达。【结论】HAS2和PTGS2在牦牛不同时期卵巢中的表达具有显著差异性，推测HAS2和PTGS2可能参与牦牛生殖生理过程的调控。 |
| 关键词: 卵丘扩展 卵巢 生殖生理 qRT-PCR Western blotting 牦牛 |
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| 基金项目:国家自然科学基金项目（31972760）；甘肃省教育厅产业支撑引导项目（2019C-03）；甘肃省教育厅创新能力提升项目（2019B-081） |
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| Expression and localization of HAS2 and PTGS2 in yak ovary at different stages |
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GAO Zechuan,ZHAO Ling,PAN Yangyang,et al
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| Abstract: |
| 【Objective】This study investigated the localization and expression of hyaluronate synthase 2 (HAS2) and prostaglandin endoperoxide synthase 2 (PTGS2) in yak ovaries at different stages to provide basis for further research of fertility.【Method】The ovaries of healthy yaks at different stages (follicular stage,luteal phase and pregnancy stage) were collected and expression levels of HAS2 and PTGS2 gene and protein were detected by quantitative Real-time PCR(qRT PCR) and Western blotting.The distribution of HAS2 and PTGS2 in ovaries at different stages was detected by immunohistochemistry (IHC).【Result】The expression of HAS2 gene in corpus luteum was significantly higher than that in follicular phase and pregnancy (P<0.05),and that in follicular phase was significantly higher than that in pregnancy (P<0.05).The expression of PTGS2 gene in luteal phase was significantly higher than that in other two periods (P<0.05),and that in follicular phase was significantly higher than that in pregnancy (P<0.05).The expression of HAS2 protein was significantly higher in luteal phase than in other two stages (P<0.05),and that in pregnancy was significantly higher than that in follicular phase (P<0.05).The expression of PTGS2 protein in luteal phase was significantly higher than that in other two stages (P<0.05),and that in follicular phase was significantly higher than that in pregnancy (P<0.05).IHC test showed that HAS2 and PTGS2 were expressed in luteal cells,granulosa layer and cumulus cells.【Conclusion】The expression of HAS2 and PTGS2 was different in yak ovary.It is speculated that HAS2 and PTGS2 may be involved in regulation of reproductive physiological process in yak. |
| Key words: cumulus expansion ovary reproductive physiological qRT-PCR Western blotting Bos grunniens |
