Yao Wang, Pan Zengxiang, Du Xing, Zhang Jinbi, Liu Honglin, Li Qifa
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing, 210095, China.
J Anim Sci Biotechnol. 2021 Oct 7;12(1):103. doi: 10.1186/s40104-021-00626-7.
Follicular atresia has been shown to be strongly associated with a low follicle utilization rate and female infertility, which are regulated by many factors such as microRNAs (miRNAs), which constitute a class of noncoding RNAs (ncRNAs). However, little is known about long noncoding RNAs (lncRNAs), which constitute another ncRNA family that regulate follicular atresia.
A total of 77 differentially expressed lncRNAs, including 67 upregulated and 10 downregulated lncRNAs, were identified in early atretic follicles compared to healthy follicles by RNA-Sequencing. We characterized a noncoding RNA that was highly expressed in atretic follicles (NORHA). As an intergenic lncRNA, NORHA was one of the upregulated lncRNAs identified in the atretic follicles. To determine NORHA function, RT-PCR, flow cytometry and western blotting were performed, and the results showed that NORHA was involved in follicular atresia by influencing GC apoptosis with or without oxidative stress. To determine the mechanism of action, bioinformatics analysis, luciferase reporter assay and RNA immunoprecipitation assay were performed, and the results showed that NORHA acted as a 'sponge', that directly bound to the miR-183-96-182 cluster, and thus prevented its targeted inhibition of FoxO1, a major sensor and effector of oxidative stress.
We provide a comprehensive perspective of lncRNA regulation of follicular atresia, and demonstrate that NORHA, a novel lncRNA related to follicular atresia, induces GC apoptosis by influencing the activities of the miR-183-96-182 cluster and FoxO1 axis.
卵泡闭锁已被证明与低卵泡利用率和女性不孕密切相关，其受多种因素调控，如构成一类非编码RNA（ncRNA）的微小RNA（miRNA）。然而，对于构成另一个调控卵泡闭锁的ncRNA家族的长链非编码RNA（lncRNA），人们了解甚少。
通过RNA测序，与健康卵泡相比，在早期闭锁卵泡中总共鉴定出77种差异表达的lncRNA，包括67种上调的和10种下调的lncRNA。我们鉴定了一种在闭锁卵泡中高表达的非编码RNA（NORHA）。作为一种基因间lncRNA，NORHA是在闭锁卵泡中鉴定出的上调lncRNA之一。为了确定NORHA的功能，进行了逆转录聚合酶链反应（RT-PCR）、流式细胞术和蛋白质免疫印迹分析，结果表明NORHA通过影响颗粒细胞（GC）凋亡参与卵泡闭锁，无论有无氧化应激。为了确定其作用机制，进行了生物信息学分析、荧光素酶报告基因检测和RNA免疫沉淀检测，结果表明NORHA作为一个“海绵”，直接与miR-183-96-182簇结合，从而阻止其对氧化应激的主要传感器和效应器FoxO1的靶向抑制。
我们提供了lncRNA对卵泡闭锁调控的全面观点，并证明NORHA是一种与卵泡闭锁相关的新型lncRNA，它通过影响miR-183-96-182簇和FoxO1轴的活性诱导GC凋亡。
