鸡环状RNA circSESN1的表达特性探究

doi:10.11843/j.issn.0366-6964.2022.07.011

摘要/Abstract

摘要： 本研究旨在鉴定鸡体内存在circSESN1的表达，并进一步探究circSESN1在鸡胸肌中的表达特性。试验一，分别选取不同生长发育阶段(E10、E19、21 d、49 d，n =4)的乌鸡组织样，利用RT-PCR、Sanger测序和qRT-PCR等技术鉴定环状RNA circSESN1，并检测circSESN1及其来源基因SESN1的时空表达特性；试验二，分别选取肉鸡腹腔注射葡萄糖(18 d，n=6)、丙酮酸钠(21 d，n =6)或胰岛素(24 d，n =6)不同时间点后的胸肌组织，探究外源性刺激对circSESN1表达的影响；试验三，选取15%能量限饲和15%蛋白限饲的肉鸡(21 d，n=10)胸肌组织，探究限饲对circSESN1表达的影响；此外，利用生物信息学分析软件对circSESN1的潜在性功能进行预测分析。结果如下：1) circSESN1在鸡体中存在表达，并且具有高度稳定性，不能被RNase R酶消解。2) circSESN1和来源基因SESN1在21 d乌鸡胸肌中的相对表达量最高，其次是腿肌和胰腺(P<0.01)。circSESN1在乌鸡胸肌不同生长发育阶段中表达量不同，在21 d时最高，在E10最低(P<0.01)。SESN1在E19乌鸡胸肌中表达量最高，在E10中最低(P<0.01)。3)外源胰岛素注射后，胸肌中circSESN1在不同时间点的表达量呈现先上升后降低的变化趋势，在注射胰岛素120 min后极显著高于基础状态或注射PBS组(P<0.01)。4)与对照组相比，15%能量限饲和15%蛋白限饲后鸡的胸肌中circSESN1的表达量显著降低(P<0.05)。5)对circSESN1潜在性功能预测后发现，circSESN1可以和多个miRNA结合，但不能编码蛋白。以上结果表明，circSESN1在鸡体内存在表达，且具有和其来源基因SESN1相似的时空表达特性。外源性胰岛素和限饲可以调控胸肌中circSESN1的表达。

关键词: 环状RNA, circSESN1, 限饲, 胰岛素, 鸡

Abstract: The purpose of this study was to identify the expression of circSESN1 in chickens and to further explore the expression characteristics of circSESN1 in chicken pectoral muscle. In experiment 1, the expression of circRNA of chicken SESN1(circSESN1) was identified by RT-PCR, Sanger sequencing, and qRT-PCR. Then the spatio-temporal expression characteristics of circSESN1 and its source gene SESN1 were detected by qRT-PCR with tissue samples of silky fowls at different growth stages (E10, E19, 21 d, 49 d, n=4). In experiment 2, the effects of exogenous stimuli on the expression of circSESN1 were explored with the pectoral muscle of broilers collected at different time points after intraperitoneal injection of glucose (18 d, n=6), sodium pyruvate (21 d, n=6) or insulin (24 d, n=6). In experiment 3, 21 d (n=10) broilers were conducted with 15% energy restriction and 15% protein restriction, and the effect of feeding restriction on the expression of circSESN1 was explored in the pectoral muscle tissue of broilers. In addition, the potential function of circSESN1 was predicted by the bioinformatics analysis software. The results showed as follows:1) circSESN1 was a circular RNA with high stability in chicken and can not be digested by RNase R enzyme. 2) The relative expression levels of circSESN1 and source gene SESN1 were the highest in the pectoralis of 21 d silky fowl, followed by leg muscle and pancreas (P<0.01). The expression level of circSESN1 dramatically varied in the pectoral muscle of birds at different growth stages, with the highest level at 21 d and the lowest at E10 (P<0.01). The expression of SESN1 was highest in the pectoral muscle of silky fowl at E19 and the lowest at E10 (P<0.01). 3) After insulin injection, the expression of circSESN1 in the pectoral muscle showed a trend of first increasing and then decreasing at different time points. CircSESN1 was significantly higher at 120 min after insulin injection than the basal state or PBS group (P<0.01). 4) Compared with the control group, the expression of circSESN1 decreased significantly by 15% energy restriction and 15% protein restriction in the pectoral muscle of birds (P<0.05). 5) After predicting the potential function of circSESN1, it was found that circSESN1 functioned as efficient miRNA sponges, but can not encode proteins. The above results showed that circSESN1 was expressed in chicken, and it had similar spatio-temporal expression characteristics with its source gene SESN1. Exogenous insulin and feeding restriction can regulate the expression of circSESN1 in the pectoral muscle.

Key words: circular RNA, circSESN1, feed restriction, insulin, chicken

中图分类号:

S831.2

邵冰豪, 高林歌, 朱星浩, 张怀勇, 陈文, 黄艳群. 鸡环状RNA circSESN1的表达特性探究[J]. 畜牧兽医学报, 2022, 53(7): 2141-2151.

SHAO Binghao, GAO Linge, ZHU Xinghao, ZHANG Huaiyong, CHEN Wen, HUANG Yanqun. Expression Characteristics of Circular RNA circSESN1 in Chicken[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2141-2151.

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