奶牛乳脂代谢关键miRNAs的筛选及鉴定

doi:10.11843/j.issn.0366-6964.2022.12.011

Abstract

Abstract: This study aimed to investigate the key miRNAs and target mRNAs affecting the process of milk fat metabolism in Chinese Holstein cows. In this study, 8 Ningxia Holstein cows with extreme differences in milk fat rate in their first lactation at mid to late lactation stage(150-220 d) were selected and divided into two groups according to differences in milk fat rate: high and low milk fat groups, with 4 replicates in each group. Milk samples from the 8 cows were collected to isolate mammary epithelial cells for transcriptome sequencing and bioinformatics analysis to screen out differentially expressed miRNA-mRNA, and their correlation with milk fat rate was analyzed. The miRNAs and mRNAs expression were verified by real-time fluorescence quantitative PCR. The results showed that 8 small RNA (sRNA) libraries were sequenced to obtain 11 976 914-16 235 680 clean reads, accounting for more than 97% of the total raw reads, with Q30 reaching more than 91%. The length of sRNA fragments were mainly distributed in the range of 21-24 nt. A total of 34 differentially expressed miRNAs (16 up-regulated and 18 down-regulated) were identified between the high and low milk fat groups. GO and KEGG enrichment analysis showed that the target genes of differentially expressed miRNAs were significantly enriched in functional items of intracellular signaling transduction, single organism processes, kinase binding and phosphotransferase activity, and significantly enriched in TNF signaling pathway, MAPK signaling pathway, Ras signaling pathway, Rap1 signaling pathway and oxytocin signaling pathway. A total of 16 miRNA-target gene pairs were obtained by comprehensive miRNA-mRNA analysis. miR-1343-3p and MTM1 were significantly negatively correlated with milk fat rate, and miR-370, miR-2285cb and SRRM2 were significantly positively correlated with milk fat rate. The expression trend of differentially expressed miRNAs and mRNAs verified by RT-qPCR were consistent with the result of transcriptome sequencing. These results explored the functional mechanisms of milk fat metabolism in Holstein cows in the Ningxia region at the miRNA and mRNA levels and identified miR-370-MTM1, miR-1343-3p-DIS3L2, miR-2285cb-XLOC-122799 and miR-2387-SRRM2 interaction pairs as possible key factors regulating milk fat metabolism in cows.

Key words: miRNA, milk fat, Chinese Holstein cows, transcriptomics, miRNA-mRNA interaction

CLC Number:

S823.91

LIU Jiamin, YU Baojun, MU Tong, ZHANG Di, FENG Xiaofang, ZHANG Juan, WANG Ying, WEN Wan, GU Yaling. Screening and Identification of Key miRNAs for Milk Fat Metabolism in Dairy Cattle[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(12): 4244-4257.

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Screening and Identification of Key miRNAs for Milk Fat Metabolism in Dairy Cattle

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