水牛卵泡不同发育阶段差异表达基因分析

Analysis of Differentially Expressed Genes in Follicles of Bubalus bubalis at Dif-ferent Developmental Stages

本研究旨在利用转录组测序技术探究摩拉水牛(Bubalus bubalis)发情期不同大小卵泡的基因表达谱,并进一步了解其对卵泡发育过程中相关激素的影响。通过对摩拉水牛进行Ovsynch法同期发情处理,B超检测卵泡大小并按照大、中、小3种类型(>8 mm、4~7 mm、0~3 mm)区分,活体采集卵泡液后提取RNA送单细胞转录组测序。将获得的数据进行组间两两比较,以|log2fold change|≥1且P<0.05为标准筛选差异转录本,并取各组差异表达转录本并集共1473个转录本做表达趋势筛选,其中587个转录本符合牛卵泡发育过程中的转录调控规律,通过GO和KEGG富集分析并筛选得到了33条通路,各通路主要与细胞间相互作用、类固醇生成和激素产生、细胞分化和发育、细胞运动、代谢调节、生理适应性等过程相关。通过对通路上的基因进行蛋白质互作网络分析,进一步获得了可能影响水牛卵泡发育过程中激素调节的关键基因,如BAMBI、FST、INHA、INHBA、INHBB、GATA6和GFAP。综上,该研究结果为进一步探究水牛卵泡发育过程中的关键基因及可能的分子调控机制提供了参考。

The purpose of this study was to explore the gene expression profiles of follicles of different sizes during estrus in Murrah buffalo(Bubalus bubalis)by transcriptome sequencing technology,and to further understand their effects on related hormones during follicular development.Ovsynch method was used to synchronize estrus in Murrah buffalo.The size of follicles was detected by B-ultrasound and divided into three types:large,medium,and small(>8 mm,4~7 mm,0~3 mm).RNA was extracted from follic-ular fluid and sent to single cell transcriptome sequencing.The obtained data were compared between groups,and the differential tran-scripts were screened with|log2fold change|≥1 and P<0.05 as the standard.A total of 1473 differentially expressed transcripts from the union of each group were selected for expression trend screening,in which 587 transcripts were in line with the transcriptional regulation law during bovine follicular development.Through GO and KEGG enrichment analysis,33 pathways were screened,and each pathway was mainly related to intercellular interaction,steroid production and hormone production,cell differentiation and devel-opment,cell movement,metabolic regulation,physiological adaptability,and other processes.Through the analysis of the protein in-teraction network of the genes in the pathway,the key genes that may affect the hormone regulation during the development of buffalo fol-licles were further obtaind,such as BAMBI,FST,INHA,INHBA,INHBB,GATA6,and GFAP.In conclusion,these results provide a reference for further investigation of key genes and possible molecular regulatory mechanisms in the development of buffalo follicles.