Ding Ying, Zhao Juanjuan, Xu Xianshuai, Zuo Qisheng, Zhang Yani, Jin Kai, Han Wei, Li Bichun
Key Laboratory of Animal Genetics, Breeding and Molecular Design of Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.
Joint International Research Laboratory of Agriculture and Agri-Product Safety of the Ministry of Education of China, Yangzhou University, Yangzhou 225009, China.
Stem Cells Int. 2023 Apr 4;2023:4956871. doi: 10.1155/2023/4956871. eCollection 2023.
Autophagy plays an important role in the pluripotency and differentiation of stem cells. Transcriptome data showed that the autophagy genes and were significantly upregulated in primordial germ cells (PGCs). The Kyoto Encyclopedia of Genes and Genome (KEGG) results showed that the lysosome signaling pathway, which is related to autophagy, was significantly enriched in PGCs. Quantitative RT-PCR, western blotting, and transmission electron microscopy (TEM) results showed that autophagy was expressed in both embryonic stem cells (ESCs) and PGCs but was significantly activated in PGCs. To explore the role of autophagy in the differentiation of chicken ESCs into PGCs, autophagy was activated and inhibited using rapamycin and bafilomycin A1, respectively. Results of qRT-PCR, flow cytometry, and indirect immunofluorescence showed that the efficiency of PGC formation significantly decreased after autophagy inhibition. Our results showed, for the first time, that autophagy plays an indispensable role in the formation of chicken PGCs, which lays the foundation for studying the mechanism of autophagy in chicken PGCs and in bird gene editing and the rescue of endangered birds.
自噬在干细胞的多能性和分化中起重要作用。转录组数据显示，自噬基因在原始生殖细胞（PGCs）中显著上调。京都基因与基因组百科全书（KEGG）结果表明，与自噬相关的溶酶体信号通路在PGCs中显著富集。定量逆转录聚合酶链反应（qRT-PCR）、蛋白质免疫印迹法和透射电子显微镜（TEM）结果显示，自噬在胚胎干细胞（ESCs）和PGCs中均有表达，但在PGCs中显著激活。为了探究自噬在鸡胚胎干细胞向PGCs分化中的作用，分别使用雷帕霉素和巴弗洛霉素A1激活和抑制自噬。qRT-PCR、流式细胞术和间接免疫荧光结果显示，自噬抑制后PGC形成效率显著降低。我们的结果首次表明，自噬在鸡PGCs的形成中起不可或缺的作用，这为研究鸡PGCs中自噬的机制、鸟类基因编辑以及濒危鸟类的拯救奠定了基础。
