Acta Veterinaria et Zootechnica Sinica ›› 2022, Vol. 53 ›› Issue (1): 151-160.doi: 10.11843/j.issn.0366-6964.2022.01.015

• ANIMAL BIOTECHNOLOGY AND REPRODUCTION • Previous Articles     Next Articles

Effects of ASH1L Knockdown on mRNA Profiling in Bovine Cumulus Cells

TIAN Yaqing, CUI Lixin, HAO Haisheng, ZOU Huiying, PANG Yunwei, ZHAO Xueming, ZHU Huabin, DU Weihua*   

  1. Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing 100193, China
  • Received:2021-03-09 Online:2022-01-23 Published:2022-01-26

Abstract: The study aimed to explore the effects of absent, small, or homeotic 1-like (ASH1L) gene knockdown on mRNA profiling in bovine cumulus cell. In this study, the mRNA profiling in cumulus cells with or without interference of ASH1L gene was measured using RNA sequencing. Differentially expressed genes (DEGs) were identified, and bioinformatics analyses (differential expression of genes,function,signaling pathway,alternative splicing (AS) and SNP) were carried out. Transcriptional levels were confirmed using quantitative RT-PCR,3 replicates per trial. Results showed that a total of 2 046 DEGs were screened in cells with or without interfere-nce of ASH1L gene, of which 1 078 were up-regulated and 968 were down-regulated. Those DEGs mainly enriched into 2 802 GO terms including cell division, cell junctions and histone methyltransferase activity. Fifty-three pathways were involved by those DEGs such as leukocyte transendothelial migration, PI3K-Akt and cell adhesion molecule. The results of gene set enrichment analysis showed that ASH1L gene was closely involved in developmental induction, H3K4 specific histone methyltransferase activity, histone demethylation, etc. Moreover, 117 994 AS events with 12 patterns were identified; In the meantime,522 205 SNPs were detected and the frequency of base transitions was significantly higher than that of transversions. In summary, the mRNA profiling between bovine cumulus cells with or without interference of ASH1L gene were different. It was speculated that ASH1L gene may be involved in leukocyte transendothelial migration and cell adhesion molecule to regulate cell proliferation. Also, ASH1L can affect gene expression by AS and SNP, which may provide a theoretical basis for further study on the function of ASH1L in cumulus cell proliferation and embryo development.

Key words: ASH1L, transcriptome sequencing, cumulus cells, cattle

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