發(fā)布時(shí)間：2019-07-08 11:36

【摘要】：本試驗(yàn)旨在研究胚期注射葉酸對(duì)肉雞脾和胸腺中IGF2基因表達(dá)的影響,并進(jìn)一步探究其調(diào)控基因表達(dá)的表觀遺傳學(xué)機(jī)制。選擇重量和大小相近的種蛋360枚,隨機(jī)分為4組,在孵化期第11天(E11)分別注射0(對(duì)照)、50、100和150μg的葉酸。每個(gè)處理選取體重相近的48只1日齡雛雞,隨機(jī)分為6個(gè)重復(fù),每個(gè)重復(fù)8只雞,在飼養(yǎng)的第21和42天取胸腺和脾組織。用RT-PCR方法檢測(cè)組織中IGF2的相對(duì)表達(dá)量;重亞硫酸鹽修飾克隆測(cè)序法(BSP)檢測(cè)IGF2啟動(dòng)子區(qū)域(-648/-479bp)甲基化水平;DNase I-qPCR方法檢測(cè)IGF2啟動(dòng)子區(qū)域(-847/-616bp)染色質(zhì)構(gòu)象。結(jié)果表明,胚期注射100和150μg葉酸顯著提高了受精蛋孵化率(P0.05);胚期注射150μg葉酸顯著提高了42日齡脾中IGF2的表達(dá)量(P0.05)、降低了42日齡脾中該基因啟動(dòng)子區(qū)的甲基化水平(P0.05),并且也顯著增加了42日齡脾中IGF2啟動(dòng)子區(qū)的染色質(zhì)可接近度(P0.05);然而在21日齡時(shí)各葉酸處理組對(duì)脾中IGF2的表達(dá)均無(wú)顯著影響(P0.05)。與脾組織不同,150μg葉酸處理組顯著提高了21日齡胸腺中IGF2的表達(dá)(P0.05),對(duì)42日齡胸腺中該基因的表達(dá)卻沒(méi)有顯著影響(P0.05),并且與對(duì)照組相比,150μg葉酸處理組中21日齡胸腺組織IGF2啟動(dòng)子區(qū)的染色質(zhì)可接近度也顯著增加(P0.05)。由此可見(jiàn),在種蛋孵化第11天注射葉酸可通過(guò)影響基因啟動(dòng)子區(qū)的甲基化水平和染色質(zhì)構(gòu)象來(lái)調(diào)控組織中IGF2的表達(dá),并且存在組織器官和時(shí)間特異性。
文內(nèi)圖片：
圖片說(shuō)明：圖４胚期注射葉酸對(duì)脾中ＩＧＦ２啟動(dòng)子區(qū)甲基化的影響Ｆｉｇ．４ＥｆｆｅｃｔｓｏｆｆｏｌｉｃａｃｉｄｉｎｊｅｃｔｅｄｏｎＥ１１ｏｎｐｒｏｍｏｔｅｒ
[Abstract]:The purpose of this study was to study the effect of folic acid injection on the expression of IGF2 gene in spleen and thymic gland of broilers, and to further explore the epigenetic mechanism of folic acid injection. 360 eggs of similar weight and size were randomly divided into 4 groups and injected with 0 (control), 50100 and 150 渭 g folic acid on the 11th day of incubation (E11). In each treatment, 48 1-day-old chicks of similar weight were randomly divided into 6 replicates, 8 chickens each. The thymic gland and spleen tissue were taken on the 21st and 42nd day of feeding. The relative expression of IGF2 in tissues was detected by RT-PCR, the methylation level of IGF2 promoter region (- 648/-479bp) was detected by heavy sulfite modified clone sequencing (BSP), and the chromatin conformations of IGF2 promoter region (- 847/-616bp) were detected by DNase I-qPCR. The results showed that injection of 100 渭 g folic acid and 150 渭 g folic acid significantly increased the hatching rate of fertilized eggs (P 0.05), and 150 渭 g folic acid significantly increased the expression of IGF2 in spleen of 42 days old (P 0.05), decreased the methylation level of promoter region of IGF2 gene in 42 days old spleen (P 0.05), and also significantly increased the chromatin closeness of IGF2 promoter region of 42 days old spleen (P 0.05). However, at 21 days of age, folic acid treatment groups had no significant effect on the expression of IGF2 in spleen (P 0.05). Different from spleen tissue, 150 渭 g folic acid treatment group significantly increased the expression of IGF2 in 21-day-old thymic gland (P 0.05), but had no significant effect on the expression of IGF2 gene in 42-day-old thymic gland (P 0.05). Compared with the control group, the chromatin closeness of 21-day-old thymic tissue promoter region in 150 渭 g folic acid treatment group was also significantly increased (P 0.05). It can be seen that injection of folic acid on the 11th day of hatching can regulate the expression of IGF2 in tissues by affecting the methylated level of gene promoter region and chromatin conformational, and has tissue, organ and time specificity.
【作者單位】： 西北農(nóng)林科技大學(xué)動(dòng)物科技學(xué)院;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(31272464) 教育部新世紀(jì)優(yōu)秀人才項(xiàng)目(NCET-12-0476) 陜西省農(nóng)業(yè)攻關(guān)計(jì)劃(2014k01-18-02;2015NY149) 陜西省科技統(tǒng)籌創(chuàng)新工程計(jì)劃項(xiàng)目(2015KTCQ02-19)
【分類(lèi)號(hào)】：S831

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