Gu Yuzhou, Wu Kexin, Niu Bowen, Wang Zhiting, Jie Yuchen, Fan Zixuan, Li Junying, Sun Congjiao, Hou Zhuo-Cheng, Shao Li-Wa
State Key Laboratory of Animal Biotech Breeding, China Agricultural University, Beijing, China.
National Engineering Laboratory for Animal Breeding, College of Animal Science and Technology, China Agricultural University, Beijing, China.
Front Genome Ed. 2025 Jan 8;6:1512108. doi: 10.3389/fgeed.2024.1512108. eCollection 2024.
Primordial germ cells (PGCs) play a crucial role in transmitting genetic information to the next-generation. In chickens, genetically edited PGCs can be propagated and subsequently transplanted into recipient embryos to produce offspring with desired genetic traits. However, during early embryogenesis, the effects of external conditions on PGC migration through the vascular system to the gonads have yet to be explored, which may affect the efficiency of preparing gene-edited chickens. In this study, we investigated the effects of hyperthermia on the biological characteristics and migration of chicken PGCs. A gonad-derived PGC line of White Leghorn (WLH) chicken was established and verified through PAS staining and immunofluorescence of PGC-specific proteins. To visually observe PGC migration , GFP-positive PGCs were prepared and locations of chimeras were validated. Cell viability, glycogen granule contents, and mRNA expression levels of pluripotency markers ( and ), germ cell-specific markers ( and ), and telomerase reverse transcriptase () were reduced in PGCs cultured under high temperatures (43°C for 12, 24, and 48 h). After the heat treatment of donor PGCs (43°C) or recipient embryos (39.5°C), GFP-positive PGCs in gonads were rarely observed. Taken together, our results underscore the negative effects of hyperthermia on the biological characteristics and migration of chicken PGCs, which provides valuable insights for the implementation of PGC-based gene editing techniques in chickens.
原始生殖细胞（PGCs）在将遗传信息传递给下一代的过程中起着至关重要的作用。在鸡中，经过基因编辑的PGCs可以进行增殖，随后移植到受体胚胎中，以产生具有所需遗传特征的后代。然而，在胚胎早期发育过程中，外部条件对PGCs通过血管系统迁移至性腺的影响尚未得到探索，这可能会影响制备基因编辑鸡的效率。在本研究中，我们调查了高温对鸡PGCs生物学特性和迁移的影响。通过对PGC特异性蛋白进行PAS染色和免疫荧光，建立并验证了白来航鸡（WLH）性腺来源的PGC系。为了直观地观察PGC迁移，制备了绿色荧光蛋白（GFP）阳性的PGCs，并验证了嵌合体的位置。在高温（43°C，处理12、24和48小时）条件下培养的PGCs中，细胞活力、糖原颗粒含量以及多能性标志物（和）、生殖细胞特异性标志物（和）和端粒酶逆转录酶（）的mRNA表达水平均降低。在对供体PGCs（43°C）或受体胚胎（39.5°C）进行热处理后，性腺中很少观察到GFP阳性的PGCs。综上所述，我们的结果强调了高温对鸡PGCs生物学特性和迁移的负面影响，这为在鸡中实施基于PGC的基因编辑技术提供了有价值的见解。
