外源注射miR-499-5p对肉鸡腓肠肌转录组的影响

doi:10.11843/j.issn.0366-6964.2020.09.007

Abstract

Abstract: The objective of this study was to investigate the influence of miR-499-5p injection on gastrocnemius muscle fiber diameter and gene expression, and improve the molecular regulation mechanism of miR-499-5p in skeletal muscle development in broiler. Sixteen healthy Daheng cocks aged 14 days were randomly assigned into 2 groups with 8 replicates per group. agomiR-NC(5 nmol) were injected into the chicks in control group, and agomiR-miR-499-5p(5 nmol) were injected into the chicks in treating group, respectively. agomiR-NC and agomiR-miR-499-5p were injected once every 3 days for a total of 6 times. The gastrocnemius muscle in the 2 groups were collected after slaughter for the measurement of muscle fiber diameter and RNA-seq analysis. The results showed that the muscle fiber diameter of gastrocnemius of chicks in treating group was significantly shorter than that of chicks in control group (P<0.05). 308 263 164 and 316 696 152 clean reads were obtained from the treating group and control group, respectively. The ratios of these reads to chicken reference genome were 90.29% and 89.57%, respectively. Compared with the chicks in the control group, there were 18 differentially expressed genes in gastrocnemius muscle of miR-499-5p treated chicks, of which 12 up-regulated genes and 6 down-regulated genes (P<0.05). The top 2 differentially expressed genes were FOS proto oncogene (FOS) and early growth response gene-1 (EGR1). GO enrichment analysis showed that the differentially expressed genes were mainly enriched in the transmembrane receptor protein serine/threonine kinase signaling pathway, transcriptive regulation, lipid biosynthetic process and skeletal muscle regeneration. KEGG analysis showed that the differentially expressed genes were mainly involved in the MAPK signaling pathway and Wnt signaling pathway. A total of 78 target genes of miR-499-5p were predicted by bioinformatics software. GO enrichment analysis showed that the 78 target genes were also mainly enriched in the negative regulation of protein serine/threonine kinase activity and transcription factor activity. In conclusion, the results showed that miR-499-5p could change the gastrocnemius muscle fiber diameter and affect characteristics of muscle fiber. FOS, EGR1, CYR61 and WNT7A, which were significantly associated with the alteration of muscle fiber diameter, might be involved in the regulation of fat deposition and muscle development through the pathways mentioned above in chicks after miR-499-5p injection.

Key words: broiler, diameter of muscle fiber, transcriptome sequencing, muscle development

CLC Number:

S831.2

LI Zhixiong, XU Yaou, LIN Yaqiu, YANG Chaowu, YU Chunlin, WU Jinbo, CHEN Ling. Effect of miR-499-5p Injection on Transcriptome in Gastrocnemius Muscle of Broiler[J]. Acta Veterinaria et Zootechnica Sinica, 2020, 51(9): 2098-2108.

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Effect of miR-499-5p Injection on Transcriptome in Gastrocnemius Muscle of Broiler

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