Zhang H, Jin P F, Niu L L, Li L, Wang L J, Chen Y, Zhang G J, Zhang H P, Zhong T
Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, College of Animal Science and Technology, Sichuan Agricultural University, Chengdu, China.
Institute of Nanjiang Yellow Goat Breeding Science, Bazhong, China.
Genet Mol Res. 2016 Sep 2;15(3):gmr8678. doi: 10.4238/gmr.15038678.
DIO3 gene encoding type 3 iodothyronine deiodinase is an imprinted gene, located in the DLK1-DIO3 (delta-like 1 homolog-type 3 iodothyronine deiodinase) imprinted domain, and is potentially involved in degrading excessive amounts of thyroid hormone to protect embryogenesis. However, the underlying regulatory mechanism of the imprinted DIO3 gene expression during fetal and neonatal development in goats has not been elucidated. In this study, we explored the DNA methylation patterns of the caprine DIO3 intragenic CpG island and quantified gene expression level in six tissues from Chinese Nanjiang Yellow 3-day old kids. The expression of the DIO3 gene was determined using quantitative reverse transcription-polymerase chain reactions (qRT-PCRs), while the identification of methylation patterns was determined using bisulfite-sequencing PCRs. Modest, and non-significant (P > 0.05), methylation patterns were noted for the DIO3 CpG island methylation in the brain, heart, liver, kidney, lung, and longissimus dorsi tissues (ranging from 26.48 to 34.92%). The expression level of the DIO3 mRNA was significantly higher (P < 0.05) in the liver tissue than in the other five tissues. Pearson's correlation analysis revealed that there was no significant relationship between methylation and gene expression (P > 0.05), which indicated that the expression of the caprine DIO3 gene was likely modified by other regulatory elements. This study identified DNA methylation and expression patterns of the DIO3 gene in goats and provided insights into further regulatory mechanisms of expression and imprinting in the DLK1-DIO3 domain.
编码3型碘甲状腺原氨酸脱碘酶的DIO3基因是一个印记基因，位于DLK1 - DIO3（δ样1同源物 - 3型碘甲状腺原氨酸脱碘酶）印记区域，可能参与降解过量的甲状腺激素以保护胚胎发育。然而，山羊胎儿和新生儿发育过程中印记的DIO3基因表达的潜在调控机制尚未阐明。在本研究中，我们探究了中国南江黄羊3日龄羔羊六个组织中DIO3基因内CpG岛的DNA甲基化模式，并对基因表达水平进行了定量分析。使用定量逆转录 - 聚合酶链反应（qRT - PCR）测定DIO3基因的表达，同时使用亚硫酸氢盐测序PCR确定甲基化模式。在脑、心脏、肝脏、肾脏、肺和背最长肌组织中，DIO3 CpG岛甲基化呈现适度且无显著差异（P > 0.05）的模式（范围为26.48%至34.92%）。DIO3 mRNA的表达水平在肝脏组织中显著高于其他五个组织（P < 0.05）。Pearson相关性分析表明甲基化与基因表达之间无显著关系（P > 0.05），这表明山羊DIO3基因的表达可能受其他调控元件的影响。本研究确定了山羊DIO3基因的DNA甲基化和表达模式，并为DLK1 - DIO3区域表达和印记的进一步调控机制提供了见解。
