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2025年8月2日 星期六
农业生物技术学报  2019, Vol. 27 Issue (5): 773-784    DOI: 10.3969/j.issn.1674-7968.2019.05.002
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
鸡小黄卵泡环状RNA的鉴定和功能分析
沈曼曼1, 2, 李婷婷1, 陈兰1, 吴鹏飞1, 陈福祥1, 张跟喜1, 王金玉1, *
1 扬州大学 动物科学与技术学院,扬州 225009;
2 江苏省家禽科学研究所,扬州 225216
Identification and Functional Analysis of circRNAs in Chicken (Gallus gallus) Small Yellow Follicle
SHEN Man-Man1, 2, LI Ting-Ting1, CHEN Lan1, WU Peng-Fei1, CHEN Fu-Xiang1, ZHANG Gen-Xi1, WANG Jin-Yu1, *
1 College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China;
2 Jiangsu Institute of Poultry Science, Yangzhou 225216, China
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摘要 鸡(Gallus gallus)卵泡膜和卵泡颗粒细胞与哺乳动物的功能不同,鸡卵泡发育过程中最重要的是小黄卵泡(small yellow follicle, SYF)的发育。为研究鸡小黄卵泡环状RNA (circular RNA, circRNA)的表达水平,丰富鸡的卵泡发育理论,本研究以京海黄鸡小黄卵泡颗粒细胞(granulosa cells of SYF, SYFG)和膜细胞(theca cells of SYF, SYFT)为实验对象,去除总RNA中的线性RNA,然后通过Illumina Hi-Seq 4000测序平台进行测序,经生物信息学分析初步筛选出差异表达的circRNAs。然后利用基因本体(Gene Ontoloty, GO)和京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes, KEGG)对差异表达circRNAs的母源基因进行功能注释和富集分析。结果显示,SYFG和SYFT中共鉴定出9 904个circRNAs,其中4 199个circRNAs在两种细胞中均广泛表达,在膜细胞中表达数量更高;特征分析发现,circRNAs在染色体上广泛分布,染色体越长则分布越多,circRNA长度主要分布在100~400 nt之间,平均长度为296 nt;共有625个circRNAs在两种细胞中差异表达,其中在SYFG中上调表达271个,在SYFT中上调表达354个;在SYFG中上调表达的circRNAs参与细胞增殖、细胞识别、脂肪酸合成、脂类代谢等过程,经分析得到候选基因TOX高迁移率蛋白家族成员2 (TOX high mobility group box family member 2, TOX2)、胰岛素样生长因子受体1 (insulin like growth factor 1 receptor, IGF1R)和磷脂酰肌醇-4,5-二磷酸3-激酶催化亚单位β (phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit β, PIK3CB);在SYFT中上调表达的circRNAs集中于核苷酸结合、粘着吸附、促性腺激素释放激素(gonadotropin releasing hormone, GnRH)等信号通路,经分析得到候选基因单胺氧化酶A(monoamine oxidase A, MAOA)、成纤维细胞生长因子受体(fibroblast growth factor receptor 2, FGFR2)和丝裂原活化蛋白激酶1 (mitogen-activated protein kinase 1, MAPK1)。综上所述,本研究筛选出的circRNAs可能主要通过与线性转录本竞争或作为竞争性内源RNA (competing endogenous RNA, ceRNA)来发挥功能,本研究结果从circRNA角度丰富了鸡卵泡发育的基础遗传理论。
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沈曼曼
李婷婷
陈兰
吴鹏飞
陈福祥
张跟喜
王金玉
关键词 京海黄鸡卵泡颗粒细胞膜细胞环状RNA竞争性内源RNA    
Abstract:The function of follicular theca cells and granulosa cells of chicken (Gallus gallus) is different from that of mammals.The most important process in chicken follicular development is the development of small yellow follicle (SYF).In order to study the expression level of circular RNA (circRNA) in chicken follicle and enrich follicular development theory in chickens, the present study used SYF granulosa cells (SYFG) and SYF theca cells (SYFT) from Jinghai Yellow chicken as samples.RNA-seq libraries with linear RNA removed were prepared and sequenced with Hi-Seq 4000 platform instrument, and differentially expressed circRNAs were preliminarily screened by bioinformatics analysis.Then Gene ontoloty (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis for differentially expressed circRNAs were performed to find the putative functional annotation and enrichment of the host genes.Lastly, a total of 9 904 circRNAs in SYFG and SYFT were identified with 4 199 pervasively distributed in the two kinds of cells.The number of circRNAs in SYFT was greater than that in SYFG.When coming to the feature of circRNAs, it was found that the most length of circRNAs spanned from 100 to 400 nt with an average length of 296 nt, and they distributed on all chromosome 1~28 and sex chromosome with more distribution in longer chromosome.After differentially expressed analysis, 625 dysregulated circRNAs were found with 271 up-regulated in SYFG and 354 up-regulated in SYFT.The up-regulated circRNAs in SYFG might involve in cell proliferation, cell recognition, fatty acid biosynthesis, and metabolic process, and the candidate genes such as TOX high mobility group box family member 2 (TOX2), insulin like growth factor 1 receptor (IGF1R),and phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit β (PIK3CB) potentially affect granulosa development as circRNAs isoform.The up-regulated circRNAs in SYFT might participate in nucleotide binding, focal adhesion, and several pathways such as gonadotropin releasing hormone (GnRH) signaling pathway, apoptosis, oocyte meiosis associated with reproduction, and candidate genes as monoamine oxidase A (MAOA), fibroblast growth factor receptor 2 (FGFR2) and mitogen-activated protein kinase 1 (MAPK1) were selected.In conclusion, the circRNAs screened in present study might play their roles mainly by competing with linear transcripts or as competitive endogenous RNA (ceRNA).The results of present study enrich the basic genetic theory of chicken follicular development in the perspective of circRNA.
Key wordsJinghai Yellow chicken    Follicle    Granulosa cell    Theca cell    circRNA    ceRNA
收稿日期: 2018-12-05     
ZTFLH:  S831.2  
基金资助:国家现代农业产业技术体系(CARS-41)、江苏高校优势学科建设工程和江苏省自然科学基金(No.BK20181217)
通讯作者: Jinyuwang@yzu.edu.cn   
引用本文:   
沈曼曼, 李婷婷, 陈兰, 吴鹏飞, 陈福祥, 张跟喜, 王金玉. 鸡小黄卵泡环状RNA的鉴定和功能分析[J]. 农业生物技术学报, 2019, 27(5): 773-784.
SHEN Man-Man, LI Ting-Ting, CHEN Lan, WU Peng-Fei, CHEN Fu-Xiang, ZHANG Gen-Xi, WANG Jin-Yu. Identification and Functional Analysis of circRNAs in Chicken (Gallus gallus) Small Yellow Follicle. 农业生物技术学报, 2019, 27(5): 773-784.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2019.05.002     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2019/V27/I5/773
 
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