猪circ0015292的鉴定及其在卵泡中的表达研究

doi:10.16431/j.cnki.1671-7236.2019.03.014

《中国畜牧兽医》 ›› 2019, Vol. 46 ›› Issue (3): 756-763.doi: 10.16431/j.cnki.1671-7236.2019.03.014

猪circ0015292的鉴定及其在卵泡中的表达研究

谢苏¹, 李梦寻^1,2, 邱梅玉¹, 公红斌¹, 孙义姗¹, 孙晓梅¹, 黄涛¹

1. 石河子大学动物科技学院, 石河子 832000;
2. 华中农业大学动物科技学院动物遗传育种与繁殖教育部重点实验室, 武汉 430070

收稿日期:2018-08-30 出版日期:2019-03-20 发布日期:2019-03-20
通讯作者: 黄涛 E-mail:taohuang100@sina.com
作者简介:谢苏(1993-),男,湖南新邵人,硕士生,研究方向:动物遗传育种与繁殖,E-mail:249107362@qq.com
基金资助:
国家自然科学基金（31460586）

Identification of Porcine circ0015292 and Its Expression Pattern in Ovarian Follicles

XIE Su¹, LI Mengxun^1,2, QIU Meiyu¹, GONG Hongbin¹, SUN Yishan¹, SUN Xiaomei¹, HUANG Tao¹

1. College of Animal Science and Technology, Shihezi University, Shehezi 832000, China;
2. Key Laboratory of Animal Breeding and Reproduction of Minstry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China

Received:2018-08-30 Online:2019-03-20 Published:2019-03-20

摘要/Abstract

摘要：

试验旨在通过高通量测序及生物信息学分析circRNA在猪卵泡发育过程中的作用及其可能的机制，为进一步探索circRNA对猪卵泡发育的调控机理奠定生物学基础。采集梅山和杜洛克猪不同级别的卵泡及各组织样本，提取总RNA，采用RNA-Seq技术对卵泡中差异表达的circRNA进行筛选，同时验证并分析其在梅山猪和杜洛克猪卵泡中的表达情况，并用实时荧光定量PCR进行组织表达谱分析。结果显示：试验证实了circ0015292的存在，且其在肌肉、心脏、肝脏、脾脏、肺脏、肾脏、卵巢、输卵管、子宫、垂体及下丘脑中均有不同程度的表达，在心脏、脾脏、肺脏、卵巢中的表达量相对较高；circ0015292在梅山猪S卵泡中的表达量极显著高于杜洛克猪（P<0.01），在杜洛克猪M1、M2、L卵泡中的表达量均显著高于梅山猪（P<0.05）；其潜在靶基因GADD45G、CCR2、IL20RB、CFL1、LAMB3与猪的卵泡发育相关。综合试验结果，circ0015292在不同组织、不同时期卵泡中均有不同水平的表达；部分靶标基因参与细胞周期及卵巢发育相关信号通路，提示其可能通过对靶基因的调控作用间接参与卵泡发育过程，这为猪繁殖机理的研究提供了新思路。

关键词: 猪; circ0015292; 组织表达谱; RNA-Seq

Abstract:

High throughput sequencing and bioinformatics were used to explore the role of circRNA in porcine follicular development and its possible mechanism,in order to provide the biological foundation for further exploring the regulatory mechanism of circRNA on porcine follicular development.Different levels of follicles and tissue samples from Meishan and Duroc pigs were collected,and then the total RNA was extracted.The differential expression of circRNA in follicles was screened by RNA-Seq technology.Meanwhile,the expression of RNA-Seq in the follicles of Meishan and Duroc pigs was validated and analyzed.At the same time,Real-time PCR was used to make tissue expression profiles.The results confirmed the existence of circ0015292,and it was expressed differently in muscle,heart,liver,spleen,lung,kidney,ovary,fallopian tube,uterus,pituitary gland and hypothalamus,and the expression level was relatively high in heart,spleen,lung and ovary.The expression level of circ0015292 in follicle S of Meishan pigs was extremely significantly higher than that of Duroc pigs (P<0.01),and the expression levels of follicles M1,M2 and L in Duroc pigs were significantly higher than those in Meishan pigs (P<0.05).The potential target genes of circ0015292 (GADD45G,CCR2,IL20RB,CFL1,LAMB3) were related to the development of porcine follicles.The results indicated that circ0015292 was expressed at different levels in different tissues and follicles at different stages;some target genes were involved in cell cycle and ovarian development-related signaling pathways,suggesting that circ0015292 might indirectly participate in follicular development through the regulation of target genes.This result would provide a new idea for the study of pig reproductive mechanism.

Key words: sow; circ0015292; tissue expression profile; RNA-Seq

中图分类号:

S813.3

谢苏, 李梦寻, 邱梅玉, 公红斌, 孙义姗, 孙晓梅, 黄涛. 猪circ0015292的鉴定及其在卵泡中的表达研究[J]. 《中国畜牧兽医》, 2019, 46(3): 756-763.

XIE Su, LI Mengxun, QIU Meiyu, GONG Hongbin, SUN Yishan, SUN Xiaomei, HUANG Tao. Identification of Porcine circ0015292 and Its Expression Pattern in Ovarian Follicles[J]. , 2019, 46(3): 756-763.

参考文献

[1] 于红.表观遗传学:生物细胞非编码RNA调控的研究进展[J].遗传,2009,31(11):1077-1086. YU H.Epigenetics:Progress in the regulation of non coding RNA in biological cells[J].Genetics,2009,31(11):1077-1086.(in Chinese)
[2] PLACE R F,NOONAN E J.Non-coding RNAs turn up the heat:An emerging layer of novel regulators in the mammalian heat shock response[J].Cell Stress Chaperones,2014,19(2):159-172.
[3] LASDA E,PARKER R.Circular RNAs:Diversity of form and function[J].RNA,2014,20:1829-1842.
[4] SANGER H L,KLOTZ G,RIESNER D,et al.Viroids are single-stranded covalently closed circular RNA molecules existing as highly base-paired rod-like structures[J].Proceeding of the National Academy of Sciences,1976,73:3852-3856.
[5] KOS A,DIJKEMA R,ARNBERG A C,et al.The hepatitis δ virus possesses a circular RNA[J].Nature,1986,323(6088):558-560.
[6] JECK W R,SORRENTINO J A,WANG K,et al.Circular RNAs are abundant,conserved,and associated with ALU repeats[J].RNA,2013,19(2):141-157.
[7] 李培飞,陈声灿,邵永富,等.环状RNA的生物学功能及其在疾病发生中的作用[J].生物物理学报,2014,30(1):15-23. LI P F,CHEN S C,SHAO Y F,et al.Biological function of cyclic RNA and its role in the pathogenesis of diseases[J].Journal of Biophysics,2014,30(1):15-23.(in Chinese)
[8] MEMCZAK S,JENS M,ELEFSINIOTI A,et al. circular RNAs are a large class of animal RNAs with regulatory potency[J].Nature,2013,495(7441):333-338.
[9] SALZMAN J,CHEN R E,OLSEN M N,et al.Cell-type specific features of circular RNA expression[J].PLoS Genetics,2013,9(9):e1003777.
[10] 宋新强,张丽丽.环状RNA的生物学功能[J].生物学教学,2016,41:72-73. SONG X Q,ZHANG L L.Biological functions of cyclic RNA[J].Biology Teaching,2016,41:72-73.
[11] HANSEN T B,JENSEN T I,CLAUSEN B H,et al.Natural RNA circles function as efficient microRNA sponges[J].Nature,2013,495(7441):384-388.
[12] HANSEN T B,KJEMS J,DAMGAARD C K.circular RNA and miR-7 in cancer[J].Cancer Research,2013,73(18):5609-5612.
[13] MORTEN T V,THOMAS B,HANSEN S T,et al.Spatio-temporal regulation of circular RNA expression during porcine embryonic brain development[J].Genome Biology,2015,16:245,1-17.
[14] LIVAK K J,SCHMITTGEN T D.Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method[J].Methods,2001,25(4):402-408.
[15] XUAN L,QU L,HAN Z,et al.circular RNA:A novel biomarker for progressive laryngeal cancer[J].American Journal of Translational Research,2016,8(2):932.
[16] WANG P L,BAO Y,YEE M C,et al.circular RNA is expressed across the eukaryotic tree of life[J].PLoS One,2014,9(6):e90859.
[17] VENØ M T,HANSEN T B,VEN S T,et al.Spatio-temporalregulation of circular RNA expression during porcine embryonicbrain development[J].Genome Biology,2015,16(1):245.
[18] SUN J,XIE M,HUANG Z,et al.Integrated analysis of non-coding RNA and mRNA expression profiles of 2 pig breeds differing in muscle traits[J].Journal of Animal Science,2017,95(3):1092-1103.
[19] LIANG G,YANG Y,NIU G,et al.Genome-wide profiling of Sus scrofa circular RNAs across nine organs and three developmental stages[J].DNA Research,2017,24(1):1-13.
[20] SHI F,LAPOLT P S.Relationship between FoxO1 protein levels and follicular development,atresia,and luteinization in the rat ovary[J].Journal of Endocrinology,2003,179(2):195-203.
[21] 丁威.二花脸猪卵泡形成和早期发育的特性及其相关通路的研究[D].南京:南京农业大学,2010. DING W.Characteristics and related pathways of follicle formation and early development in Erhualian pigs[D].Nanjing:Nanjing Agricultural University,2010.

猪circ0015292的鉴定及其在卵泡中的表达研究

Identification of Porcine circ0015292 and Its Expression Pattern in Ovarian Follicles

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