本文選題：山羊胎兒 + TET��； 參考：《中國農(nóng)業(yè)科學》2017年14期

【摘要】：【目的】通過檢測TET1和Wnt信號通路相關(guān)基因以及DKK家族基因在山羊胎兒發(fā)育早期的表達變化,分析TET1與Wnt通路基因的相關(guān)性,為TET1調(diào)控山羊胎兒發(fā)育研究提供理論依據(jù)�！痉椒ā窟x取12只健康大足黑山羊母羊,自然發(fā)情后與同一只種公羊自然交配。采用剖腹產(chǎn)手術(shù)的方法,分別獲得妊娠20、25、30、60和90d的胎兒,對胎兒的生長指標(體重、體長)進行統(tǒng)計,并采集了60和90d胎兒的組織器官樣品(心、肝、肺、腎、腦、皮膚),通過Real-time PCR(RT-PCR)檢測各樣品中TET1基因,DKK家族基因(DKK1、DKK2、DKK3)和Wnt家族基因(Wnt2、Wnt2b、Wnt4、Wnt5a、Wnt5b、Wnt7b、Wnt16)的相對表達量。利用SPSS軟件分析山羊胎兒發(fā)育早期不同階段TET1與WNT信號通路相關(guān)基因相關(guān)性以及基因表達顯著性(P0.05)。【結(jié)果】山羊妊娠早期胎兒生長在60d后有顯著變化。熒光定量檢測結(jié)果表明,TET1基因表達隨妊娠天數(shù)的增加呈上升趨勢。Wnt家族基因在山羊胎兒發(fā)育中都檢測到表達(Wnt2,-2b,-4,-5a,-5b,-7b,-16)。其中,Wnt2和Wnt7b表達量隨胎兒發(fā)育逐漸增高;Wnt2b、Wnt5a、Wnt5b、Wnt7b在妊娠30 d時有顯著高表達(P0.05);Wnt4在胎兒發(fā)育20 d時表達顯著(P0.05);Wnt16基因在妊娠25 d有顯著高表達(P0.05)。DKK家族基因表達檢測結(jié)果顯示,DKK1在胎兒發(fā)育早期階段都有表達,DKK2/3在妊娠初期表達量較低,后期表達增高。通過組織中基因表達檢測顯示,TET1在90d胎兒肝、肺、腎和腦中的表達水平相比于60d胎兒組織升高,肝中表達量顯著(P0.05)。Wnt家族基因Wnt2在組織器官中有相對活躍的表達,妊娠90d胎兒肺中表達量極顯著(P0.01);Wnt16基因在胎兒皮膚組織中表達顯著(P0.05),且維持在一個較高的水平;Wnt5a和Wnt7b在腎中表達顯著(P0.05),其他Wnt基因在組織中都有表達。相關(guān)性分析顯示,胎兒生長指標(體重、體長)變化與TET1的表達呈極顯著正相關(guān)(P0.01);TET1在胎兒發(fā)育早期的表達與Wnt2、Wnt7b、Wnt16呈現(xiàn)正相關(guān),與Wnt2b、Wnt4、Wnt5a、Wnt5b呈負相關(guān),其中與Wnt5b呈顯著負相關(guān)(P0.05),與Wnt7b呈極顯著正相關(guān)(P0.01)。Wnt通路基因之間也有相互關(guān)系,Wnt2與Wnt4呈極顯著負相關(guān)(P0.01)。Wnt2與Wnt7b,Wnt2b與Wnt5a、Wnt5b,Wnt5a與Wnt5b呈極顯著正相關(guān)(P0.01)。Wnt4與Wnt5a呈顯著正相關(guān)(P0.05)�！窘Y(jié)論】獲得了TET1與Wnt基因在山羊胎兒發(fā)育早期的表達模式,并進行了相關(guān)性分析,填補了這些基因在山羊方面的研究空白;TET1與Wnt基因?qū)ι窖蛱涸缙诘陌l(fā)育和組織的形成是一個動態(tài)的調(diào)控變化過程;TET1基因表達與部分Wnt基因呈現(xiàn)顯著正相關(guān),部分呈現(xiàn)顯著負相關(guān);Wnt通路基因之間表達量呈現(xiàn)一定的相關(guān)性。這些數(shù)據(jù)為TET1與Wnt分子調(diào)控山羊早期胎兒發(fā)育的機制深入研究提供了參考。
[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.
【作者單位】： 西南大學動物科技學院/重慶市牧草與草食家畜重點實驗室/重慶市草食動物資源保護與利用工程技術(shù)研究中心;
【基金】：國家級大學生創(chuàng)新創(chuàng)業(yè)訓練計劃(201510635013) 重慶高校創(chuàng)新團隊建設(shè)計劃資助項目(CXTDG201602004) 中央高�；緲I(yè)務(wù)費專項資金(XDJK2017A003) 重慶市社會民生科技創(chuàng)新專項項目(cstc2016shmszx80064)
【分類號】：S827

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