鸡环状RNA circWWP1的鉴定及组织表达研究

doi:10.16431/j.cnki.1671-7236.2023.10.005

摘要/Abstract

摘要： 【目的】鉴定鸡体内存在环状RNA(circRNA)含WW结构域的E3泛素蛋白连接酶1(circWWP1)的表达,并进一步探究其组织表达特征及其在鸡肌肉生长发育过程中的作用及可能作用机制,为研究circWWP1对鸡肌肉发育的调控机制奠定基础。【方法】利用PCR扩增、DNA测序、RNase R酶消化、放线菌素D处理鉴定circWWP1的真实性和稳定性,采用实时荧光定量PCR检测circWWP1的组织表达特征。利用在线软件预测circWWP1 miRNA的结合位点及其靶基因,利用Cytoscape 3.9.1软件构建circWWP1-miRNA-mRNA网络图,并进行GO功能和KEGG通路富集分析。【结果】鸡circWWP1真实存在且稳定表达,耐受RNase R酶的消化和放线菌素D处理。circWWP1在成年鸡各组织中广泛表达,胸肌中表达量显著高于其他组织(P＜0.05);仅在22日龄雏鸡胸肌和腿肌组织中表达;成年鸡胸肌、腿肌组织中circWWP1表达量均显著高于22日龄雏鸡(P＜0.05)。circWWP1-miRNA-mRNA网络调控分析获得3个靶向miRNA(gga-miR-454-3p、gga-miR-130a-3p和gga-miR-301b-3p)及其下游靶向的208个基因;GO功能富集显示,circWWP1靶基因显著富集于38个GO条目;KEGG通路富集分析结果显示,circWWP1靶基因主要参与和细胞增殖、胚胎发育等过程相关的Hedgehog信号通路。【结论】circWWP1在鸡体内存在且稳定表达,在不同年龄鸡胸肌、腿肌组织中高表达,可能通过Hedgehog信号通路在鸡胚发育及肌肉细胞增殖、分化过程中发挥作用。

关键词: 鸡; 环状RNA; circWWP1; 组织表达; 肌肉发育

Abstract: 【Objective】 The purpose of the experiment was to identify the expression of circular RNA (circRNA) WW domain-containing E3 ubiquitin protein ligase 1 (circWWP1) in chickens,and to further explore its tissue expression characteristics,its role in chicken muscle growth and development and its possible mechanism,so as to lay a foundation for the study of the regulatory mechanism of circWWP1 on chicken muscle development.【Method】 The authenticity and stability of circWWP1 were determined by PCR amplification,DNA sequencing,RNase R enzyme digestion and actinomycin D treatment,and the tissue expression characteristics of circWWP1 were detected by Real-time quantitative PCR.The circWWP1 miRNA binding site and its target genes were predicted by online software.The circWWP1-miRNA-mRNA network was constructed by Cytoscape 3.9.1 software,and GO function and KEGG pathway enrichment analysis were performed.【Result】 circWWP1 was present and stably expressed in chickens,which were resistant to RNase R enzyme digestion and actinomycin D treatment.circWWP1 was widely expressed in all tissues of adult chickens,and the expression level in breast muscle was significantly higher than that in other tissues (P＜0.05).It was only expressed in breast muscle and leg muscle of 22-day-old chicks.The expression level of circWWP1 in breast muscle and leg muscle of adult chickens was significantly higher than that of 22-day-old chicks (P＜0.05).The analysis of circWWP1-miRNA-mRNA network regulation obtained three target miRNAs (gga-miR-454-3p,gga-miR-130a-3p and gga-miR-301b-3p) and their downstream targets of 208 genes.GO function enrichment showed that the circWWP1 target genes were significantly enriched in 38 GO items.KEGG pathway enrichment analysis showed that circWWP1 target genes were mainly involved in Hedgehog signaling pathway related to cell proliferation and embryonic development.【Conclusion】 circWWP1 existed and was stably expressed in chickens,and was highly expressed in breast muscle and leg muscle tissues of chickens at different ages,which might play a role in chick embryo development and muscle cell proliferation and differentiation through Hedgehog signaling pathway.

Key words: chicken; circRNA; circWWP1; tissue expression; muscle development

中图分类号:

S831

杨雅玲, 沈雪梅, 喻世刚, 王钢, 古丽帕日·艾克拜, 刘武军. 鸡环状RNA circWWP1的鉴定及组织表达研究[J]. 中国畜牧兽医, 2023, 50(10): 3917-3927.

YANG Yaling, SHEN Xuemei, YU Shigang, WANG Gang, GULIPARI Aikebai, LIU Wujun. Identification and Tissue Expression of Circular RNA circWWP1 in Chicken[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(10): 3917-3927.

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