Cheng Juanru, Xing Qinghua, Pan Yu, Yang Yanyan, Zhang Ruimen, Shi Deshun, Deng Yanfei
Guangxi Key Laboratory of Animal Breeding and Disease Control, College of Animal Science and Technology, Guangxi University, Nanning 530004, China.
Chongqing Academy of Animal Sciences, Chongqing 402460, China.
Int J Mol Sci. 2024 Dec 27;26(1):153. doi: 10.3390/ijms26010153.
The specific expression profile and function of circular RNA (circRNA) in follicular atresia remain largely unknown. Here, the circRNA expression profiles of granulosa cells derived from healthy follicles (HFs) and antral follicles (AFs) in buffalo were analyzed by RNA-seq, and the mechanism of a differentially expressed circRNA (DEcircRNA) circTEC regulating the granulosa cell function that affects follicular atresia was further explored. RNA-seq results showed that a total of 112 DEcircRNAs were identified. Among them, circTEC was highly expressed in HF, and its circular structure was confirmed by RNase R digestion assay, reversed PCR and Sanger sequencing. Functional experiments demonstrated that circTEC promotes the proliferation and steroid hormone synthesis of buffalo granulosa cells (bGCs), and it also inhibits their apoptosis. In-depth mechanism analysis showed that the expression level of circTEC in bGCs from AFs was adversely related to miR-144-5p and consistent with FZD3. CircTEC acts as an endogenous sponge of miR-144-5p to regulate the expression of the target gene FZD3 in AFs, which promotes the proliferation of bGCs and inhibits bGCs apoptosis, thereby inhibiting follicular atresia in buffalo. In summary, our study revealed the regulatory role of the circTEC/miR-144-5p/FZD3 axis during follicular atresia in buffalo. These results provided new insights into the biological mechanism underlying follicular atresia.
环状RNA(circRNA)在卵泡闭锁中的具体表达谱和功能仍不清楚。本研究通过RNA测序分析了水牛健康卵泡(HF)和腔前卵泡(AF)来源的颗粒细胞的circRNA表达谱,并进一步探讨了差异表达的circRNA(DEcircRNA)circTEC调控颗粒细胞功能从而影响卵泡闭锁的机制。RNA测序结果显示,共鉴定出112个DEcircRNA。其中,circTEC在HF中高表达,通过RNase R消化试验、逆转PCR和Sanger测序证实了其环状结构。功能实验表明,circTEC促进水牛颗粒细胞(bGCs)的增殖和类固醇激素合成,并抑制其凋亡。深入的机制分析表明,AF来源的bGCs中circTEC的表达水平与miR-144-5p呈负相关,与FZD3呈正相关。circTEC作为miR-144-5p的内源性海绵,调节AF中靶基因FZD3的表达,促进bGCs的增殖并抑制bGCs凋亡,从而抑制水牛卵泡闭锁。总之,我们的研究揭示了circTEC/miR-144-5p/FZD3轴在水牛卵泡闭锁过程中的调控作用。这些结果为卵泡闭锁的生物学机制提供了新的见解。