Ministry of Education Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources, College of Fisheries and Life Sciences, Shanghai Ocean University, 999 Hucheng Huan Road, Shanghai 201306, China.
Department of Biological Sciences, National University of Singapore, Science Drive 4, Singapore 117543, Singapore.
Sci Rep. 2016 Jul 13;6:29658. doi: 10.1038/srep29658.
Primordial germ cell (PGC) specification early in development establishes the germline for reproduction and reproductive technologies. Germline replacement (GR) is a powerful tool for conservation of valuable or endangered animals. GR is achievable by germ cell transplantation into the PGC migration pathway or gonads. Blastula cell transplantation (BCT) can also lead to the chimeric germline containing PGCs of both donor and host origins. It has remained largely unknown whether BCT is able to achieve GR at a high efficiency. Here we report efficient GR by BCT into blastula embryos in the fish medaka (Oryzias latipes). Specifically, dnd depletion completely ablated host PGCs and fertility, and dnd overexpression remarkably boosted PGCs in donor blastulae. BCT between normal donor and host produced a germline transmission rate of ~4%. This rate was enhanced up to ~30% upon PGC boosting in donors. Most importantly, BCT between PGC-boosted donors and PGC-ablated hosts led to more than 90% fertility restoration and 100% GR. Therefore, BCT features an extremely high efficiency of fertility recovery and GR in medaka. This finding makes medaka an ideal model to analyze genetic and physiological donor-host compatibilities for BCT-mediated surrogate production and propagation of endangered lower vertebrates and biodiversity.
原始生殖细胞 (PGC) 早期发育中的特化建立了生殖系，用于繁殖和生殖技术。生殖系替代 (GR) 是保护有价值或濒危动物的有力工具。通过将生殖细胞移植到 PGC 迁移途径或性腺中，可以实现 GR。囊胚细胞移植 (BCT) 也可以导致含有供体和宿主起源的 PGC 的嵌合生殖系。BCT 是否能够以高效率实现 GR 仍然很大程度上未知。在这里，我们报告了通过 BCT 高效地实现鱼类斑马鱼 (Oryzias latipes) 囊胚胚胎中的 GR。具体来说，dnd 耗竭完全消除了宿主 PGCs 和生育能力，而 dnd 过表达显著增加了供体囊胚中的 PGCs。正常供体和宿主之间的 BCT 产生了约 4%的生殖系传递率。在供体中 PGC 增强的情况下，该比率提高到约 30%。最重要的是，在 PGC 增强的供体和 PGC 耗竭的宿主之间的 BCT 导致超过 90%的生育力恢复和 100%的 GR。因此，BCT 在斑马鱼中具有极高的生育力恢复和 GR 效率。这一发现使斑马鱼成为分析遗传和生理供体-宿主相容性的理想模型，用于 BCT 介导的替代生产和濒危低等脊椎动物和生物多样性的繁殖。
