DNA甲基化调控牛AQP1基因的胎盘特异性印记

为揭示牛AQP1 (aquaporin 1)基因在不同组织及胎盘中的印记状态,以及DNA甲基化修饰在印记中的调控机制,本研究采用基于SNP的PCR产物直接测序的方法,对32头健康雌性成年荷斯坦奶牛心组织及15个自然分娩后的胎盘试验样本进行检测,确定了5头杂合子个体牛和3个杂合子胎盘,对其组织(心、肝、脾、肺、肾、肌肉和...

DNA Methylation Regulate the Genomic Imprinting of AQP1 Gene Specific in Bovine Placenta

The study aimed to explore the imprinting status of AQP1 gene in various tissues and placenta of bovine and to determine the role of DNA methylation in regulating imprinted expression of AQP1 gene. The direct sequencing of RT-PCR products was used based on SNP, 5 heterozygous cows and 3 heterozygous placentas were identified from the heart tissues of 32 healthy female adult Holstein cows and 15 placentas after natural delivery, respectively. The allele expression and imprinting status of AQP1 gene in 7 tissues(heart, liver, spleen, lung, kidney, muscle and fat) and placentas of heterozygous cattle were analyzed. The DNA methylation status of the CpG island located the promoter and exon 1 of AQP1 gene was analyzed in heart, liver, two placentas and corresponding sperms by bisulfite sequencing. The results showed that AQP1 gene was in biallelic expression in 7 tissues tested of heterozygous cows. In the placenta, AQP1 gene was in maternal monoallelic expression by analyzing the parental genotype of heterozygous placenta. Furthermore, analysis of the DNA methylation status of CpG island in AQP1 gene promoter in tissues, placenta and corresponding sperms of bovine showed that there were no differentially methylated regions (DMR) in the biallele expressed heart and liver tissues, but in placenta with monoallelic expression, there were DMR, while in sperm with paternal allele expression, there was hyper-methylation. These results suggest that AQP1 gene is a paternal imprinting gene, and DNA methylation modification of the AQP1 CpG island is involved in the regulation of imprinting expression in bovine placenta.The expression of AQP1 gene is biallelic in the tested tissues.