基于高通量测序的梅花鹿卵巢衰老的转录组分析

doi:10.16431/j.cnki.1671-7236.2023.08.013

Abstract

Abstract: 【Objective】 This study was aimed to obtain the transcriptome information of ovary tissues in young and old sika deer,and provide a basis for further study the potential mechanism of ovarian aging in sika deer.【Method】 The ovaries of 4 years young sika deer and 10 years old sika deer were used as test samples.The samples were processed with paraffin sectioning,hematoxylin-eosin staining and follicle counting.RNA was extracted from the ovarian samples using the Trizol method,and a transcriptome library was constructed.The Illumina HiSeq^TM 2000 sequencing platform was used to sequence,the sequencing results were analyzed for differentially expressed genes,GO function,KEGG pathway and protein interaction network analysis.Selected differentially expressed genes were further analyzed using Real-time quantitative PCR.The expression of trimethylation of lysine 4 on histone H3 protein subunit (H3K4me3) protein in ovaries of young and old sika deer were detected by Western blotting analysis using RIPA lysis solution.【Result】 The histological analysis results showed that the number of ovarian follicles in old sika deer was significantly lower than that in young sika deer (P＜0.05),and the number of atresia follicles in old sika deer was extremely significantly higher than that of young sika deer (P＜0.01).Transcriptome analysis showed that there were 1 477 differentially expressed genes of ovaries between old and young sika deer,of which 503 genes were upregulated and 974 genes were downregulated.GO function and KEGG pathway enrichment analysis showed that the immune system process,transcription,DNA damage repair,inflammatory response,apoptosis and other biological events were related to ovarian aging.Real-time quantitative PCR results showed that there were 17 genes with significant or extremely significant expression of ovaries between old and young sika deer (P＜0.05 or P＜0.01),indicating that RNA-Seq results were reliable.Western blotting analysis results showed that the expression of H3K4me3 protein in ovaries was significantly different between old and young sika deer (P＜0.05).【Conclusion】 The number of follicles in ovaries of young and old sika deer showed a significant difference.The PI3K-Akt signaling pathway had been identified to affect the efficiency of DNA damage repair and participate in ovarian aging of sika deer via RNA-Seq.The high expression of H3K4me3 in ovaries of old sika deer was related to transcription activation.

Key words: sika deer; ovarian aging; RNA-Seq; H3K4me3

CLC Number:

S825

LIN Lefeng, DIAO Yunfei, FAN Bingfeng, LIU Lixiang, SHAO Jing, ZHAO Xiangyuan, XU Baozeng. Transcriptome Analysis of Aging Sika Deer (Cervus nippon) Ovaries Based on High-throughput Sequencing[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(8): 3157-3170.

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Transcriptome Analysis of Aging Sika Deer (Cervus nippon) Ovaries Based on High-throughput Sequencing

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