山羊早期胎儿组织TET1与Wnt通路基因的表达变化及其相关性

发布时间：2018-07-11 15:41

本文选题：山羊胎儿 + TET　；参考：《中国农业科学》2017年14期

【摘要】：【目的】通过检测TET1和Wnt信号通路相关基因以及DKK家族基因在山羊胎儿发育早期的表达变化,分析TET1与Wnt通路基因的相关性,为TET1调控山羊胎儿发育研究提供理论依据。【方法】选取12只健康大足黑山羊母羊,自然发情后与同一只种公羊自然交配。采用剖腹产手术的方法,分别获得妊娠20、25、30、60和90d的胎儿,对胎儿的生长指标(体重、体长)进行统计,并采集了60和90d胎儿的组织器官样品(心、肝、肺、肾、脑、皮肤),通过Real-time PCR(RT-PCR)检测各样品中TET1基因,DKK家族基因(DKK1、DKK2、DKK3)和Wnt家族基因(Wnt2、Wnt2b、Wnt4、Wnt5a、Wnt5b、Wnt7b、Wnt16)的相对表达量。利用SPSS软件分析山羊胎儿发育早期不同阶段TET1与WNT信号通路相关基因相关性以及基因表达显著性(P0.05)。【结果】山羊妊娠早期胎儿生长在60d后有显著变化。荧光定量检测结果表明,TET1基因表达随妊娠天数的增加呈上升趋势。Wnt家族基因在山羊胎儿发育中都检测到表达(Wnt2,-2b,-4,-5a,-5b,-7b,-16)。其中,Wnt2和Wnt7b表达量随胎儿发育逐渐增高;Wnt2b、Wnt5a、Wnt5b、Wnt7b在妊娠30 d时有显著高表达(P0.05);Wnt4在胎儿发育20 d时表达显著(P0.05);Wnt16基因在妊娠25 d有显著高表达(P0.05)。DKK家族基因表达检测结果显示,DKK1在胎儿发育早期阶段都有表达,DKK2/3在妊娠初期表达量较低,后期表达增高。通过组织中基因表达检测显示,TET1在90d胎儿肝、肺、肾和脑中的表达水平相比于60d胎儿组织升高,肝中表达量显著(P0.05)。Wnt家族基因Wnt2在组织器官中有相对活跃的表达,妊娠90d胎儿肺中表达量极显著(P0.01);Wnt16基因在胎儿皮肤组织中表达显著(P0.05),且维持在一个较高的水平;Wnt5a和Wnt7b在肾中表达显著(P0.05),其他Wnt基因在组织中都有表达。相关性分析显示,胎儿生长指标(体重、体长)变化与TET1的表达呈极显著正相关(P0.01);TET1在胎儿发育早期的表达与Wnt2、Wnt7b、Wnt16呈现正相关,与Wnt2b、Wnt4、Wnt5a、Wnt5b呈负相关,其中与Wnt5b呈显著负相关(P0.05),与Wnt7b呈极显著正相关(P0.01)。Wnt通路基因之间也有相互关系,Wnt2与Wnt4呈极显著负相关(P0.01)。Wnt2与Wnt7b,Wnt2b与Wnt5a、Wnt5b,Wnt5a与Wnt5b呈极显著正相关(P0.01)。Wnt4与Wnt5a呈显著正相关(P0.05)。【结论】获得了TET1与Wnt基因在山羊胎儿发育早期的表达模式,并进行了相关性分析,填补了这些基因在山羊方面的研究空白;TET1与Wnt基因对山羊胎儿早期的发育和组织的形成是一个动态的调控变化过程;TET1基因表达与部分Wnt基因呈现显著正相关,部分呈现显著负相关;Wnt通路基因之间表达量呈现一定的相关性。这些数据为TET1与Wnt分子调控山羊早期胎儿发育的机制深入研究提供了参考。
[Abstract]:[Objective] to analyze the correlation between TET1 and Wnt pathway gene by detecting the expression of TET1 and Wnt signaling pathway related genes and DKK family gene in the early development of goat fetus, and to provide a theoretical basis for the study of TET1 regulation of goat fetal development. [Methods] 12 healthy black goat ewes were selected, after natural estrus and the same one The fetus of 20,25,30,60 and 90d pregnancy was obtained by caesarean section. The growth index (weight, body length) of the fetus was counted, and the tissues and organs of 60 and 90d fetuses (heart, liver, lung, kidney, brain, skin) were collected and the TET1 gene and DKK family gene (D) were detected by Real-time PCR (RT-PCR). KK1, DKK2, DKK3) and the relative expression of Wnt family genes (Wnt2, Wnt2b, Wnt4, Wnt5a, Wnt5b, Wnt7b, Wnt16). The results of light quantitative detection showed that the expression of TET1 gene was increased with the increase of the number of gestation days. The expression of.Wnt family gene was detected in the development of goat fetus (Wnt2, -2b, -4, -5a, -5b, -7b, -16). The expression of Wnt2 and Wnt7b increased with fetal development. The expression of the fetus at 20 d was significant (P0.05), and the expression of Wnt16 gene at 25 D in pregnancy was significantly higher (P0.05).DKK family gene expression detection results showed that DKK1 was expressed in the early stage of fetal development, DKK2/3 was expressed in the early stage of pregnancy, and the expression increased in the early stage of pregnancy. The expression level in the brain was higher than that of 60d fetal tissue, and the expression of.Wnt gene in the liver was significantly (P0.05).Wnt gene Wnt2 expressed in the tissues and organs, and the expression of 90d in the fetal lung was very significant (P0.01); the Wnt16 gene expressed significantly in the fetal skin (P0.05) and maintained at a higher level; Wnt5a and Wnt7b were in the kidney. The expression of P0.05 and other Wnt genes were expressed in the tissues. The correlation analysis showed that the change of fetal growth index (body weight, body length) was positively correlated with the expression of TET1 (P0.01), and the expression of TET1 in the early development of the fetus was positively correlated with Wnt2, Wnt7b, Wnt16, and was negatively correlated with Wnt2b, Wnt4, Wnt5a, Wnt5b. Negative correlation (P0.05) has a very significant positive correlation with Wnt7b (P0.01).Wnt pathway genes, and Wnt2 and Wnt4 have a significant negative correlation (P0.01).Wnt2 and Wnt7b, Wnt2b and Wnt5a, Wnt5b, and a significant positive correlation. [Conclusion] the gene is obtained in the goat fetus The expression patterns of early development and correlation analysis fill the gap of these genes in goats. TET1 and Wnt genes are a dynamic regulation and regulation process for the early development and tissue formation of goat fetuses; TET1 gene expression has a significant positive correlation with some Wnt genes, and some of them have significant negative correlation; Wnt There is a certain correlation between the expression level of these genes. These data provide a reference for further research on the mechanism of TET1 and Wnt molecular regulation of early fetal development in goats.
【作者单位】：西南大学动物科技学院/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;
【基金】：国家级大学生创新创业训练计划(201510635013) 重庆高校创新团队建设计划资助项目(CXTDG201602004) 中央高校基本业务费专项资金(XDJK2017A003) 重庆市社会民生科技创新专项项目(cstc2016shmszx80064)
【分类号】：S827

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