鸡不同组织及细胞中gga-miR-103-3p 的表达分析及其关键靶基因筛选

doi:10.15933/j.cnki.1004-3268.2024.11.016

摘要/Abstract

摘要： ：为初步阐明gga-miR-103-3p对鸡脂肪沉积和肌肉发育的调控作用，采用荧光定量PCR（qPCR）检测gga-miR-103-3p在S3系鸡和隐性白羽鸡（RR）腹部脂肪、肝脏、腿肌及腹部脂肪细胞、肌内脂肪细胞、成肌细胞中的表达情况，并筛选其关键靶基因。结果表明，gga-miR-103-3p在肝脏、腹部脂肪、腿肌及腹部脂肪细胞、肌内脂肪细胞、成肌细胞中均表达，并存在组织和品种（系）差异性。在肝脏中，ggamiR-103-3p在0周龄（0W）S3系鸡中的表达量显著高于RR鸡，对于S3系鸡，16周龄（16W）时gga-miR-103-3p的表达量最高，显著高于0W、2周龄（2W）、8周龄（8W）和14周龄（14W）；对于RR鸡，16W和14W时gga-miR-103-3p的表达量显著高于0W和8W。在腹部脂肪中，gga-miR-103-3p在0W RR鸡中的表达量显著高于S3系鸡。2个品种（系）鸡在0W时gga-miR-103-3p的表达量均显著高于2W、8W、14W和16W，S3系鸡的表达量随着发育时期的推进而下降。在腿肌中，gga-miR-103-3p在16W时RR鸡中的表达量显著高于S3系鸡，在0W时则相反；S3系鸡中gga-miR-103-3p的表达量随着发育时期的推进总体上下降，在0W时表达量显著高于其他周龄；RR鸡中gga-miR-103-3p的表达量随着发育时期的推进呈现先下降后升高的趋势，16W时显著高于其他周龄。在腹部脂肪细胞和肌内脂肪细胞中，gga-miR-103-3p在分化4 d和分化6 d的表达量均显著高于增殖期；在成肌细胞中，gga-miR-103-3p的表达量随分化时间的延长而升高，分化5 d和分化7 d时的表达量均显著高于增殖期。gga-miR-103-3p靶基因的GO、KEGG富集及蛋白质互作分析结果表明，FBXW7、CDK6、NF1和PIK3R1是gga-miR-103-3p的重要靶基因，其中FBXW7和PIK3R1基因尤为关键。综上，gga-miR-103-3p可能是通过FBXW7、CDK6、NF1和PIK3R1影响肌肉发育、脂肪沉积，其中FBXW7和PIK3R1基因尤为关键。

关键词: 鸡, 脂肪沉积, 肌肉发育, gga-miR-103-3p, 靶基因

Abstract: In order to preliminarily elucidate the regulatory role of gga‑miR‑103‑3p in fat deposition and muscle development in chickens，fluorescence quantitative PCR（qPCR）was used to detect the expression of gga‑miR‑103‑3p in abdominal fat，liver，leg muscle，abdominal adipocytes，intramuscular adipocytes and adult myoblasts in chickens of the S3 lineage and recessive white‑feathered（RR）chickens，and its key target genes were screened.The results showed that gga‑miR‑103‑3p was expressed in liver，abdominal fat，leg muscle，abdominal adipocytes，intramuscular adipocytes and adult myoblasts，and there were differences among tissues and breed（line）.In the liver，gga‑miR‑103‑3p expression level was significantly higher in S3 line chickens than that in RR chickens at 0 week of age（0W）.For S3 chickens，gga‑miR‑103‑3p expression level was the highest at 16 weeks of age（16W），which was significantly higher than those at 0W，2 weeks of age（2W），8 weeks of age（8W）and 14 weeks of age（14W）；For RR chickens，gga‑miR‑103‑3p expression level was significantly higher at 16W and 14W than those at 0W and 8W. In abdominal fat，gga‑miR‑103‑3p expression level was significantly higher in RR chickens than that in S3 chickens at 0W. gga‑miR‑103‑3p expression level was significantly higher at 0W than those at 2W，8W，14W and 16W in S3 and RR chickens，and decreased in S3 chickens with the advancement of developmental period.In the leg muscle，the expression level of gga‑miR‑103‑3p was significantly higher in RR chickens than that in S3 chickens at 16W，and vice versa at 0W；The expression level of gga‑miR‑103‑3p in S3 chickens generally declined with the advancement of developmental period，and the expression level was significantly higher at 0W than those at other weeks of age；The expression level of gga‑miR‑103‑3p in RR chickens first declined and then increased with the advancement of developmental period，and the expression level was significantly higher at 16W than those at other weeks of age. In abdominal adipocytes and intramuscular adipocytes，the expression levels of gga‑miR‑103‑3p at 4 d of differentiation and 6 d of differentiation were significantly higher than that at the proliferative stage；In adult myoblasts，the expression level of gga‑miR‑103‑3p increased with the extension of differentiation，and the expression levels of gga‑miR‑103‑3p were significantly higher at 5 d of differentiation and 7 d of differentiation than that at the proliferative stage.The GO，KEGG enrichment and protein interaction analyses of gga‑miR‑103‑3p target genes showed that FBXW7，CDK6，NF1 and PIK3R1 were important target genes of gga‑miR‑103‑3p，of which FBXW7 and PIK3R1 were particularly critical.In conclusion，gga‑miR‑103‑3p may affect muscle development and fat deposition through FBXW7，CDK6，NF1 and PIK3R1，of which FBXW7 and PIK3R1 are particularly critical.

Key words: Chicken, Fat deposition, Muscle development, gga?miR?103?3p, Target gene

中图分类号:

S831

黄华云, 李瑞瑞, 杨苗苗, 赵振华, 王钱保, 梁忠, 黄正洋, 吴兆林, 李春苗, 韩威. 鸡不同组织及细胞中gga-miR-103-3p 的表达分析及其关键靶基因筛选[J]. 河南农业科学, 2024, 53(11): 147-155.

HUANG Huayun, LI Ruirui, YANG Miaomiao, ZHAO Zhenhua, WANG Qianbao, LIANG Zhong, HUANG Zhengyang, WU Zhaolin, LI Chunmiao, HAN Wei. Expression Analysis of gga‑miR‑103‑3p in Different Tissues and Cells in Chicken and Screening of Its Key Target Genes[J]. Journal of Henan Agricultural Sciences, 2024, 53(11): 147-155.

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