中国畜牧兽医2024，Vol.51Issue(7)：2751-2765,15.DOI:10.16431/j.cnki.1671-7236.2024.07.003
基于高叶酸小鼠睾丸转录组筛选牛繁殖力关键同源基因
Screening of Key Homologous Genes for Bovine Fertility Based on High Folic Acid Mouse Testicular Transcriptome
摘要
Abstract
[Objective]Based on transcriptome data of folic acid mice testis,the homologous genes of folic acid regulating bovine and mouse fertility were screened,which provided a theoretical foundation for improving male animal reproductive capacity through folic acid.[Method]Twenty four healthy and similarly growing 11 weeks old C57BL/6 male mice were randomly divided into three groups(n=8):CON,A,and B groups,fed with 2,7,and 15 mg/kg folic acid,respectively,for 57 consecutive days.After feeding,the weight of the left and right testes and seminal vesicles,sperm density,and the proportion of slow forward sperm were measured in three groups of mice.The transcriptome of mouse testes was sequenced using Illumina Novaseq™ 6000 high-throughput sequencing.The sequencing results were analyzed using differentially expressed genes(DEGs),GO functional annotation,and protein-protein interaction network(PPI)analysis to screen for key genes regulating semen quality and reproductive capacity by folic acid,and combined with homologous genes and quantitative trait genes related to reproductive capacity in Holstein bulls.Quantitative trait loci(QTLs)data were used to further screen for homologous genes that regulated male reproductive capacity in cattle and mice through folate regulation.[Result]Compared with CON group,the weight of the left and right seminal vesicles of mice in B group was extremely significantly increased(P＜0.01),and the unfavorable trait of the proportion of sperm slow foward in the semen of mice in both A and B groups were extremely significantly reduced(P＜0.01).Through RNA-Seq analysis,it was found that 423 DEGs were screened in A vs CON group,and 661 DEGs were screened in B vs CON group.A total of 85 upregulated DEGs and 40 downregulated DEGs were found in both A vs CON and B vs CON groups.Through GO functional annotation analysis,it was found that the DEGs in A vs CON group were significantly enriched in GO entries related to folate metabolism and semen quality regulation,while those in B vs CON group were significantly enriched in GO entries related to folate metabolism and reproductive organ development.The upregulated DEGs shared by both A vs CON and B vs CON groups were significantly enriched in folate metabolism and semen quality regulation,while the downregulated DEGs shared were significantly enriched in one-carbon metabolism and single sperm fertilization functions.Through PPI network analysis of DEGs,it was found that the selected hub genes might affect male animal fertility by affecting semen quality.At the same time,the hub genes screened in this study were all homologous genes of Holstein bulls,and homologous genes and their neighboring QTLs that affected the reproductive capacity of Holstein bulls at different concentrations of folic acid were identified,including inseminations per conception QTL and sperm motility QTL.Further comparing the hub gene with differentially expressed genes in the transcriptome of previous Holstein bull sperm,the homologous gene Serpinb1b(bovine homologous gene SERPINB1)was screened out,which regulated fertility in Holstein bulls and mice through folate regulation.[Conclusion]Feeding 7 and 15 mg/kg folic acid could reduce the proportion of slow forward sperm in semen.The Ser pinb1b gene(bovine homologous gene:SERPINB1)screened by RNA-Seq was one of the key genes for folic acid regulation of male animal fertility,which could provide reference information for the breeding and breeding of large dairy animal such as Holstein cows in the actual production of animal husbandry.关键词
繁殖力/叶酸/小鼠/荷斯坦公牛/转录组测序Key words
fertility/folic acid/mouse/Holstein bull/RNA-Seq分类
农业科技引用本文复制引用
吉虹锦,俞英,冶倩楠,李文龙,李志鹏,冯霞,米思远,史源钧,张金柠,郭健..基于高叶酸小鼠睾丸转录组筛选牛繁殖力关键同源基因[J].中国畜牧兽医,2024,51(7):2751-2765,15.基金项目
"十四五"科技部重点研发专项课题(2021YFD1200903、2021YFD1200900) （2021YFD1200903、2021YFD1200900）
国家自然科学基金国际(地区)合作项目(31961143009) （地区）
国家奶牛产业技术体系项目(CARS-36) （CARS-36）
