叶酸调控鸡脾和胸腺IGF2表达的表观遗传机制探究
发布时间：2019-07-08 11:36
【摘要】：本试验旨在研究胚期注射叶酸对肉鸡脾和胸腺中IGF2基因表达的影响,并进一步探究其调控基因表达的表观遗传学机制。选择重量和大小相近的种蛋360枚,随机分为4组,在孵化期第11天(E11)分别注射0(对照)、50、100和150μg的叶酸。每个处理选取体重相近的48只1日龄雏鸡,随机分为6个重复,每个重复8只鸡,在饲养的第21和42天取胸腺和脾组织。用RT-PCR方法检测组织中IGF2的相对表达量;重亚硫酸盐修饰克隆测序法(BSP)检测IGF2启动子区域(-648/-479bp)甲基化水平;DNase I-qPCR方法检测IGF2启动子区域(-847/-616bp)染色质构象。结果表明,胚期注射100和150μg叶酸显著提高了受精蛋孵化率(P0.05);胚期注射150μg叶酸显著提高了42日龄脾中IGF2的表达量(P0.05)、降低了42日龄脾中该基因启动子区的甲基化水平(P0.05),并且也显著增加了42日龄脾中IGF2启动子区的染色质可接近度(P0.05);然而在21日龄时各叶酸处理组对脾中IGF2的表达均无显著影响(P0.05)。与脾组织不同,150μg叶酸处理组显著提高了21日龄胸腺中IGF2的表达(P0.05),对42日龄胸腺中该基因的表达却没有显著影响(P0.05),并且与对照组相比,150μg叶酸处理组中21日龄胸腺组织IGF2启动子区的染色质可接近度也显著增加(P0.05)。由此可见,在种蛋孵化第11天注射叶酸可通过影响基因启动子区的甲基化水平和染色质构象来调控组织中IGF2的表达,并且存在组织器官和时间特异性。
文内图片：
图片说明：图４胚期注射叶酸对脾中ＩＧＦ２启动子区甲基化的影响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.
【作者单位】： 西北农林科技大学动物科技学院;
【基金】：国家自然科学基金项目(31272464) 教育部新世纪优秀人才项目(NCET-12-0476) 陕西省农业攻关计划(2014k01-18-02;2015NY149) 陕西省科技统筹创新工程计划项目(2015KTCQ02-19)
【分类号】：S831
本文编号：2511555
文内图片：
图片说明：图４胚期注射叶酸对脾中ＩＧＦ２启动子区甲基化的影响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.
【作者单位】： 西北农林科技大学动物科技学院;
【基金】：国家自然科学基金项目(31272464) 教育部新世纪优秀人才项目(NCET-12-0476) 陕西省农业攻关计划(2014k01-18-02;2015NY149) 陕西省科技统筹创新工程计划项目(2015KTCQ02-19)
【分类号】：S831
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