|circRNA landscape of non-pregnant endometrium during the estrus cycle in dairy goats
LIU Xiao-rui1, 2*, ZHANG Lei1, 2*, CUI Jiu-zeng1, YANG Li-chun1, HaAN Jin-cheng1, CHE Si-cheng1, CAO Bin-yun1, 2, LI Guang1, 2, SONG Yu-xuan1, 2
1 College of Animal Science and Technology, Northwest A&F University, Yangling 712100, P.R.China
2 Shaanxi Provincial Key Laboratory of Animal Genetics, Breeding and Reproduction, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, P.R.China
哺乳动物子宫内膜的发育是一个涉及众多的调节因素的复杂过程。环状RNA （circRNAs）是一种自然存在的内源性非编码RNA家族的重要成员之一，有研究报道circRNAs在多种生理过程中发挥着重要的调控作用。目的：本研究旨在分析奶山羊发情周期非妊娠子宫内膜组织中circRNA的表达谱。方法：采用链特异性去核糖体转录组测序技术对发情第5天和发情第15天奶山羊的非妊娠子宫内膜组织样进行circRNAs表达谱分析，筛选两个时期差异表达的circRNAs并利用RT-qPCR进行验证；然后利用生物信息学分析两个时期差异表达circRNAs的宿主基因，进而进行GO 和KEGG分析；最后利用Targetscan 7.0和miRanda网站分析circRNAs和miRNAs的靶向结合关系。结果：奶山羊发情第5天和发情第15天的非妊娠子宫内膜组织中共有2331个差异表达的circRNAs（P<0.05），RT-qPCR检测的10个circRNAs变化趋势与测序结果一致；其中circRNA1460的宿主基因Nipped-B 样蛋白（Nipped-B-like，NIPBL）和circRNA8694的宿主基因钙反应性转录因子（calcium responsive transcription factor，CARF）降低其circRNAs的转录形式的水平（P<0.05）参与子宫内膜的发育；差异表达circRNAs的宿主基因GO和KEGG分析结果显示注释到紧密连接（Tight junctions）条目上和鸟苷三磷酸酶（GTPases）通路上的circRNAs参与了奶山羊子发情周期中非妊娠子宫内膜的发育过程；另外，本研究中得到的circRNAs与公共数据库中的436个山羊miRNAs有靶向结合位点。结论：奶山羊发情第5天和发情第15天的非妊娠子宫内膜组织中存在差异的circRNAs，生物信息学分析表明一些circRNAs参与奶山羊发情周期非妊娠子宫内膜的发育过程；本研究构建了奶山羊发情周期非妊娠子宫内膜circRNAs文库，丰富了奶山羊的转录组信息，有助于我们研究奶山羊子宫内膜发育过程中的分子调控机制，为进一步提高奶山羊的胚胎着床的成功率和繁殖率提供理论依据。创新点：本研究选择发情第5天和发情第15天奶山羊的非妊娠子宫内膜组织进行circRNAs表达谱分析；采用核糖体去除和链特异性文库构建方案进行RNA-seq测序，保留了完整的circRNAs序列，对样本中几乎全部的circRNAs序列进行鉴定和分析。
Endometrial development is a complicated process involving numerous regulatory factors. Circular RNAs (circRNAs) have been known as a member of the naturally occurring non-coding RNA family, and are reportedly crucial for a variety of physiological processes. This study investigated the circRNA landscape of non-pregnant endometrium of dairy goats during estrus. Non-pregnant endometrial samples of goats at estrus day 5 (Ed5) and estrus day 15 (Ed15) were used to methodically analyze the circRNA landscape using strand-specific Ribo-Zero RNA-Seq. A total of 2 331 differentially expressed (P<0.05) circRNAs (DEciRs) between Ed5 and Ed15 were discovered in the goat endometrium. It was found that Nipped-B-like (NIPBL) and calcium responsive transcription factor (CARF) may participate in the development of the endometrium by decreasing (P<0.05) the levels of their circRNA-transcript forms. Furthermore, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses of DEciR host genes (hgDEciRs) revealed that tight junctions and GTPases may be involved in endometrial development during the estrus cycle. A total of 2 331 DEciRs were discovered in the endometrium at Ed5 and Ed15. Based on GO and KEGG enrichment analyses, it could be inferred that tight junctions and GTPases are likely to play an important role in the development of goat endometrium during the estrus cycle. This circRNA study greatly enhances our knowledge of global trends in the development of non-pregnant endometrium during the estrus cycle in goats; these results help us to better understand the molecular regulation of endometrial development in dairy goats.
Received: 29 March 2020
|Fund: This work was supported by the China Postdoctoral Science Foundation (2019M653776, 2020M673516), the National Key Research and Development Program of China (2016YFD0500508), the Natural Science Foundation of Shaanxi Province, China (2020JQ-265), and the PhD research startup foundation of Northwest A&F University, China (00400/Z109021811).
Correspondence ZHANG Lei, Tel: +86-29-87092527, Fax: +86-29-87092164, E-mail: email@example.com
|About author: * These authors contributed equally to this study.
Cite this article:
LIU Xiao-rui, ZHANG Lei, CUI Jiu-zeng, YANG Li-chun, HAN Jin-cheng, CHE Si-cheng, CAO Bin-yun, LI Guang, SONG Yu-xuan.
circRNA landscape of non-pregnant endometrium during the estrus cycle in dairy goats. Journal of Integrative Agriculture, 20(5): 1346-1358.
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