云南半细毛羊精子冷冻前后转录组表达差异分析

doi:10.16431/j.cnki.1671-7236.2022.03.019

Abstract

Abstract: 【Objective】 The purpose of this study was to study the relationship between differentially expressed gene and sperm freezing injury of Yunnan Semi-fine Wool sheep before and after sperm freezing.【Method】 The semen of three Yunnan Semi-fine Wool sheep were collected by electrical stimulation and divided into two parts.One was directly used to extract sperm RNA, and the other was cryopreserved for 7 days before sperm RNA extraction.Using the obtained sperm RNA, the mRNA transcriptomes of fresh and frozen-thawed sperm were examined by single-cell whole transcriptome sequencing, and mRNA transcriptome libraries were constructed and sequenced with Hiseq2500 sequencer, and the sequencing results were further filtered, and the differentially expressed genes were analyzed by GO function annotation and KEGG enrichment analysis.【Result】 6 semen samples had a total of 623 399 754 original sequencing sequences.After filtration, 514 313 802 (82.50%) high-quality Clean reads were obtained.The bioinformatics analysis results showed that there were 1 213 genes with significant difference before and after the freezing and thawing process.Among them, the abundance of 1 208 genes were down-regulated, and 5 genes were up-regulated after freezing and thawing.Through GO annotation and KEGG enrichment analysis, the differential genes were classified into 56 GO terms and were participate in 40 signal pathways.Their biological functions were closely related to sperm functions mainly including cellular process, metabolic process, response to stimulation, reproductive process, reproduction, membrane and antioxidant.Before and after sperm freezing, the main genes involved in inflammation, cytokine-cytokine receptor interaction and apoptosis signal pathway.【Conclusion】 The results showed that the cryoinjuries of sperm of Yunnan Semi-fine Wool sheep might be related to the biological process that the obtained differentially expressed genes were involved.In addition, the obtained differential genes might also be used as potential molecular markers to evaluate the quality of sheep frozen semen.

Key words: Yunnan Semi-fine Wool sheep; sperm; cryopreservation; transcriptome; differential expressed gene

CLC Number:

S826.8

LIANG Jiachong, LYU Chunrong, HONG Qionghua, WU Guoquan, QUAN Guobo. Analysis of Sperm Differential Transcriptome Expression Before and After Freezing in Yunnan Semi-fine Wool Sheep[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 973-981.

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Analysis of Sperm Differential Transcriptome Expression Before and After Freezing in Yunnan Semi-fine Wool Sheep

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