AANAT基因在滩羊不同繁殖时期卵巢中的转录差异及DNA甲基化程度分析

doi:10.16431/j.cnki.1671-7236.2021.01.025

Abstract

Abstract: The aim of this study was to detect the transcriptional differences of 5-hydroxytryptamine N-acetyltransferase (AANAT) gene in ovine ovarian tissue during the anestrus and breeding season (follicular stage and luteal stage) and to analyze whether the transcriptional differences were caused by changes in the degree of DNA methylation modification.In the study,non-pregnant Tan sheep whose natural environmental conditions and feeding management were consistent and the weight difference was within 0.5 kg were selected.Their ovarian tissues during the anestrus,follicular and luteal stages (3 ewes in each stage) were collected.SYBR dye method was used to carry out Quantitative real-time PCR to detect the transcription level of AANAT gene in the ovarian tissue of tan sheep at different reproductive stages.Subsequently,MethPrimer 2.0 online software was used to predict the CpG island in promoter region and the first exon region of AANAT gene,and the bisulfite sequencing (BSP) method was used to detect the methylation degree of the promoter region and the first exon region of the AANAT gene for the anestrus and follicle stage samples with transcriptional differences.The results showed that the transcriptional level of AANAT gene in the ovarian tissue of Tan sheep during anestrus was significantly lower than that in the follicular period in the breeding season (P<0.05),and there was no significant difference of transcriptional level between anestrus and luteal period (P>0.05).There was a CpG island with a length of 173 bp in the AANAT gene promoter region and a CG island with a length of 118 bp in the first exon region in the Tan sheep ovarian tissue.However there was no significant difference on the methylation degree of the single CpG site in promoter region and the first exon of the AANAT gene in Tan sheep ovary tissue,suggesting that the expression difference of the AANAT gene was regulated by other factors other than methylation modification.The results of this study might provide references for further study on the function of AANAT gene in seasonal estrus and follicular maturation.

Key words: Tan sheep; seasonal estrus; AANAT gene; transcription; DNA methylation

CLC Number:

S826

ZHANG Hui, LI Xiaoyu, DI Ran, LIU Yufang, CHU Mingxing. Analysis on Difference of Transcription and DNA Methylation Level of AANAT Gene in Ovary of Tan Sheep at Different Reproductive Stages[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(1): 226-234.

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Analysis on Difference of Transcription and DNA Methylation Level of AANAT Gene in Ovary of Tan Sheep at Different Reproductive Stages

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