中国农业科学 ›› 2021, Vol. 54 ›› Issue (4): 845-854.doi: 10.3864/j.issn.0578-1752.2021.04.015

• 畜牧·兽医·资源昆虫 • 上一篇    下一篇

TETs与细胞程序性死亡相关基因在山羊妊娠早期输卵管及子宫角的表达

赵乐(),杨海丽,李佳璐,杨永恒,张蓉,程文强,成磊,赵永聚()   

  1. 西南大学动物科学技术学院/重庆市牧草与草食家畜重点实验室/重庆市草食动物资源保护与利用工程技术研究中心,重庆 400715
  • 收稿日期:2020-03-12 接受日期:2020-07-10 出版日期:2021-02-16 发布日期:2021-02-16
  • 通讯作者: 赵永聚
  • 作者简介:赵乐,E-mail: zhaole0228@163.com
  • 基金资助:
    国家自然科学基金(31772564);国家重点研发计划(2018YFD0502000);重庆市自然科学基金重点项目(cstc 2020 jcyj-zdxm X0015);重庆市研究生科研创新项目(CYS19108);重庆市现代山地特色高效农业产业技术体系草食牲畜,2020【11】

Expression Patterns of TETs and Programmed Cell Death Related Genes in Oviduct and Uterus of Early Pregnancy Goats

ZHAO Le(),YANG HaiLi,LI JiaLu,YANG YongHeng,ZHANG Rong,CHENG WenQiang,CHENG Lei,ZHAO YongJu()   

  1. College of Animal Science and Technology, Southwest University/Chongqing Key Laboratory of Forage & Herbivore/Chongqing Engineering Research Center for Herbivores Resource Protection and Utilization, Chongqing 400715
  • Received:2020-03-12 Accepted:2020-07-10 Online:2021-02-16 Published:2021-02-16
  • Contact: YongJu ZHAO

摘要:

【目的】TET(ten-eleven translocation)家族蛋白在调控胚胎和胎盘发育等过程中具有重要作用,输卵管与子宫内膜细胞的死亡平衡会影响受精卵及早期胚胎的发育。因此探讨TETs、细胞程序性死亡相关基因和附植相关基因在妊娠早期山羊输卵管及子宫角中的表达规律及其潜在调控作用,以期为研究胚胎发育及附植的分子机制奠定基础。【方法】以合川白山羊为研究对象,采集妊娠19d(P19)山羊子宫角及输卵管组织,以空配19d(C19)为对照,利用H.E染色观察C19组和P19组山羊子宫角组织形态,利用qRT-PCR技术检测了胚胎附植相关基因(WNT5aOPNVEGFA)、TETsTET1TET2TET3)、细胞程序性死亡相关基因(凋亡相关基因(BAXBCL2Caspase9)、焦亡相关基因(GSDMDNLRP3Caspase1)和炎症相关基因(NF-kBTNF-α))的相对表达,并分析了TETs及细胞程序性死亡相关基因在输卵管和子宫角组织的表达相关性。【结果】H.E染色结果显示,在P19组中山羊子宫角的腺体增多,形态多变。qRT-PCR结果显示,OPN在P19组山羊子宫角中的表达量极显著高于C19组(P<0.01),BAX/BCL2的表达量显著增加(P<0.05),而WNT5aVEGFA表达差异不显著;TET1TET2TET3在山羊子宫角及输卵管中均有表达,在C19组和P19组山羊子宫角中TET1TET2的表达量极显著高于C19组和P19组中TET3的表达(P<0.01);在P19组山羊输卵管中TET2P<0.01)、NF-kBP<0.01)、Caspase1P<0.01)、GSDMDP<0.05)显著增高。相关性分析表明,输卵管中TET2TET3P<0.01)、NF-kBP<0.01)、Caspase9P<0.05)显著正相关;子宫角中,TET3VEGFAP<0.05)、WNT5aP<0.01)显著负相关,胚胎附植关键基因OPNCaspase1P<0.01)、NLRP3P<0.05)、GSDMDP<0.05)显著正相关,而细胞焦亡关键基因GSDMD在输卵管与子宫角中均与BaxP<0.05)、Caspase1P<0.01)、NLRP3P<0.05)呈显著正相关。【结论】在妊娠早期山羊子宫角和输卵管中TETs表达与细胞程序性死亡部分基因表达具有一定的相关性,推测TETs与细胞凋亡或焦亡的发生对输卵管、受精卵、早期胚胎及子宫的发育具有一定的调控作用。胚胎附植关键基因OPN与焦亡相关基因显著相关,暗示妊娠早期子宫角中细胞焦亡的发生可能参与胚胎附植。为研究TETs通过细胞程序性死亡相关基因在妊娠早期输卵管及子宫角中的表达规律提供了理论参考。

关键词: 山羊, 子宫角, 输卵管, 细胞程序性死亡, 基因表达

Abstract:

【Objective】The ten-eleven translocation (TET) protein family play crucial roles in embryo and placental development. The balance of cell death between oviduct and endometrial would affect the development of gametes and early embryos. In this study, the expression patterns and their potential regulatory role of TETs, programmed cell death related genes and blastocyst implantation-related genes in oviduct and uterus of goats in early pregnancy were mainly explored, so as to provide a basis for studying development of gametes and early embryos.【Method】In this study, the uterus and oviduct of Hechuan white goats during non-pregnant 19 days (C19) and pregnant 19 days (P19) were collected, and then the morphology uterine horn was showed by HE staining. Then, the genes, including blastocyst implantation-related genes (Wnt5a, OPN, VEGFA), TET family genes (TET1, TET2, TET3), programmed cell death-related genes (BAX, BCL2, Caspase9, GSDMD, NLRP3 and Caspase1) and inflammation-related genes (NF-kBTNF-α) were detected by qRT-PCR. Finally, the correlation between TETs and programmed cell death-related genes expression were analyzed.【Result】The results showed that the gland number increased, and its morphology varied in uterine horn in P19 group. Compared with C19 group, OPN gene expression and Bax/BCL2 in uterine horn tissue were significantly increased in P19 group (P<0.01), however, there was no significant difference between Wnt5a and VEGFA. TET family genes were expressed in both oviduct and uterus. The expression of TET1 and TET2 was significantly higher than TET3 in uterine horn (P<0.01). TET2 (P<0.01), NF-kB (P<0.01), Caspase1 (P<0.01), and GSDMD (P<0.05) gene were significantly higher in P19 group of oviduct tissue. Correlation analysis showed that the relationship between TET2 and TET3 (P<0.01), NF-kB (P<0.01) and Caspase9 (P<0.01) in oviduct tissue were positive correlation. The TET3 were negative correlated with VEGFA (P<0.05) and WNT5a (P<0.01). Blastocyst implantation-related genes OPN was positively correlated with Caspase1 (P<0.01), NLRP3 (P<0.05) and GSDMD (P<0.05) in uterine horn. The GSDMD was positively correlated with Bax (P<0.05), Caspase1 (P<0.01) and NLRP3 (P<0.05) in oviduct and uterus horn.【Conclusion】It was indicated that there was correlation between TETs and programmed cell death related genes. So, it was speculated that TET family genes and programmed cell death related genes played a key role in regulating the development of oviduct, zygote, early embryo and uterus. The key genes for embryo implantation of OPN were significantly correlated with pyroptosis related genes, suggesting that proptosis had a certain influence on the attachment of early embryos. These finding provided heretical references for studying the effect of TETs family genes and programmed cell death related genes on oviduct and uterine horn of early pregnancy.

Key words: goat, uterine horn, oviduct, programmed cell death, gene expression