SCU-CUHK Joint Laboratory for Reproductive Medicine, Key Laboratory of Birth Defects and Related Diseases of Women and Children (Sichuan University), Ministry of Education, Department of Pediatrics, West China Second University Hospital, Sichuan University, Chengdu, People's Republic of China.
The School of Life Science, Shandong University, Jinan, Shandong, People's Republic of China.
Reproduction. 2018 Sep;156(3):261-268. doi: 10.1530/REP-17-0681. Epub 2018 Jun 21.
Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene affect fertility in both sexes. However, the involvement of CFTR in regulating germ cell development remains largely unknown. Here, we used zebrafish model to investigate the role of CFTR in primordial germ cells (PGCs) development. We generated a frameshift mutant zebrafish line using CRISPR/Cas9 technique and investigated the migration of PGCs during early embryo development. Our results showed that loss of Cftr impairs the migration of PGCs from dome stages onward. The migration of PGCs was also perturbed by treatment of CFTRinh-172, a gating-specific CFTR channel inhibitor. Moreover, defected PGCs migration in mutant embryos can be partially rescued by injection of WT but not other channel-defective mutant mRNAs. Finally, we observed the elevation of and , key factors involved in zebrafish PGCs migration, in mutant zebrafish embryos. Taken together, the present study revealed an important role of CFTR acting as an ion channel in regulating PGCs migration during early embryogenesis. Defect of which may impair germ cell development through elevation of key factors involved in cell motility and response to chemotactic gradient in PGCs.
囊性纤维化跨膜电导调节因子（CFTR）基因突变会影响两性的生育能力。然而，CFTR 调节生殖细胞发育的作用在很大程度上尚不清楚。在这里，我们使用斑马鱼模型来研究 CFTR 在原始生殖细胞（PGCs）发育中的作用。我们使用 CRISPR/Cas9 技术产生了一个移码突变的斑马鱼品系，并研究了 PGCs 在早期胚胎发育中的迁移。我们的结果表明，Cftr 的缺失会损害 PGCs 从穹顶阶段开始的迁移。PGCs 的迁移也受到 CFTRinh-172 的干扰，CFTRinh-172 是一种门控特异性 CFTR 通道抑制剂。此外，突变体胚胎中缺陷的 PGCs 迁移可以通过注射 WT 但不是其他通道缺陷突变体 mRNAs 部分恢复。最后，我们观察到突变体斑马鱼胚胎中参与斑马鱼 PGCs 迁移的关键因子 和 的升高。总之，本研究揭示了 CFTR 作为离子通道在早期胚胎发生中调节 PGCs 迁移的重要作用。其缺陷可能通过升高 PGCs 中参与细胞运动和对趋化梯度反应的关键因子来损害生殖细胞发育。
