Qiao Yanzhao, Xiao Gengsheng, Zhu Xiaohua, Wen Jun, Bu Yonghui, Zhang Xinheng, Kong Jie, Bai Yinshan, Xie Qingmei
State Key Laboratory of Swine and Poultry Breeding Industry & Heyuan Branch, Guangdong Provincial Laboratory of Lingnan Modern Agricultural Science and Technology, College of Animal Science, South China Agricultural University, Guangzhou 510642, China.
College of Animal Science, South China Agricultural University, Guangzhou 510642, China.
Animals (Basel). 2024 Jul 30;14(15):2214. doi: 10.3390/ani14152214.
Avian primordial germ cells (PGCs) are essential in avian transgenic research, germplasm conservation, and disease resistance breeding. However, cultured PGCs are prone to fragmentation and apoptosis, regulated at transcriptional and translational levels, with N6-methyladenosine (m6A) being the most common mRNA modification. Resveratrol (RSV) is known for its antioxidant and anti-apoptotic properties, but its effects on PGCs and the underlying mechanisms are not well understood. This study shows that RSV supplementation in cultured PGCs improves cell morphology, significantly enhances total antioxidant capacity ( < 0.01), reduces malondialdehyde levels ( < 0.05), increases anti-apoptotic expression, and decreases expression ( < 0.05). Additionally, RSV upregulates the expression of m6A reader proteins and ( < 0.05). m6A methylation sequencing revealed changes in mRNA m6A levels after RSV treatment, identifying 6245 methylation sites, with 1223 unique to the control group and 798 unique to the RSV group. Combined analysis of m6A peaks and mRNA expression identified 65 mRNAs with significantly altered methylation and expression levels. Sixteen candidate genes were selected, and four were randomly chosen for RT-qPCR validation, showing results consistent with the transcriptome data. Notably, and are closely related to apoptosis, indicating potential research value. Overall, our study reveals the protective effects and potential mechanisms of RSV on chicken PGCs, providing new insight into its use as a supplement in reproductive stem cell culture.
禽类原始生殖细胞(PGCs)在禽类转基因研究、种质保存和抗病育种中至关重要。然而,培养的PGCs容易发生碎片化和凋亡,这在转录和翻译水平上受到调控,其中N6-甲基腺苷(m6A)是最常见的mRNA修饰。白藜芦醇(RSV)以其抗氧化和抗凋亡特性而闻名,但其对PGCs的影响及潜在机制尚不清楚。本研究表明,在培养的PGCs中添加RSV可改善细胞形态,显著提高总抗氧化能力(<0.01),降低丙二醛水平(<0.05),增加抗凋亡蛋白表达,并降低促凋亡蛋白表达(<0.05)。此外,RSV上调m6A阅读蛋白YTHDF1和YTHDF2的表达(<0.05)。m6A甲基化测序揭示了RSV处理后mRNA m6A水平的变化,共鉴定出6245个甲基化位点,其中1223个是对照组特有的,798个是RSV组特有的。对m6A峰和mRNA表达的联合分析确定了65个甲基化和表达水平显著改变的mRNA。选择了16个候选基因,随机挑选4个进行RT-qPCR验证,结果与转录组数据一致。值得注意的是,这些基因与凋亡密切相关,具有潜在的研究价值。总体而言,我们的研究揭示了RSV对鸡PGCs的保护作用及潜在机制,为其作为生殖干细胞培养补充剂的应用提供了新的见解。