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青鳉鱼的vasa基因对原始生殖细胞的迁移是必需的,但对其存活并非必需。

Medaka vasa is required for migration but not survival of primordial germ cells.

作者信息

Li Mingyou, Hong Ni, Xu Hongyan, Yi Meisheng, Li Changming, Gui Jianfang, Hong Yunhan

机构信息

State Key Laboratory of Freshwater Ecology and Biotechnology, Center for Developmental Biology, Institute of Hydrobiology, Chinese Academy of Sciences, Graduate School of the Chinese Academy of Sciences, Wuhan, China.

出版信息

Mech Dev. 2009 May-Jun;126(5-6):366-81. doi: 10.1016/j.mod.2009.02.004. Epub 2009 Feb 26.

Abstract

Vasa is essential for germline development. However, the precise processes in which vasa involves vary considerably in diverse animal phyla. Here we show that vasa is required for primordial germ cell (PGC) migration in the medakafish. vasa knockdown by two morpholinos led to the PGC migration defect that was rescued by coinjection of vasa RNA. Interestingly, vasa knockdown did not alter the PGC number, identity, proliferation and motility even at ectopic locations. We established a cell culture system for tracing PGCs at the single cell level in vitro. In this culture system, control and morpholino-injected gastrulae produced the same PGC number and the same time course of PGC survival. Importantly, vasa-depleted PGCs in culture had similar motility and locomotion to normal PGCs. Expression patterns of wt1a, sdf1b and cxcr4b in migratory tissues remained unchanged by vasa knockdown. By chimera formation we show that PGCs from vasa-depleted blastulae failed to migrate properly in the normal environment, whereas control PGCs migrated normally in vasa-disrupted embryos. Furthermore, ectopic PGCs in vasa-depleted embryos also retained all the PGC properties examined. Taken together, medaka vasa is cell-autonomously required for PGC migration, but dispensable to PGC proliferation, motility, identity and survival.

摘要

Vasa对于生殖细胞发育至关重要。然而,Vasa所涉及的精确过程在不同动物门类中差异很大。在此我们表明,在青鳉中,Vasa是原始生殖细胞(PGC)迁移所必需的。通过两种吗啉代寡核苷酸敲低Vasa会导致PGC迁移缺陷,而共注射Vasa RNA可挽救该缺陷。有趣的是,即使在异位位置,敲低Vasa也不会改变PGC的数量、特性、增殖和运动能力。我们建立了一种细胞培养系统,用于在体外单细胞水平追踪PGC。在该培养系统中,对照和注射了吗啉代寡核苷酸的原肠胚产生的PGC数量相同,PGC存活的时间进程也相同。重要的是,培养中Vasa缺失的PGC与正常PGC具有相似的运动能力和移动性。Vasa敲低后,迁移组织中wt1a、sdf1b和cxcr4b的表达模式保持不变。通过嵌合体形成,我们表明来自Vasa缺失囊胚的PGC在正常环境中无法正常迁移,而对照PGC在Vasa功能破坏的胚胎中正常迁移。此外,Vasa缺失胚胎中的异位PGC也保留了所有检测的PGC特性。综上所述,青鳉Vasa是PGC迁移所细胞自主需要的,但对于PGC增殖、运动能力、特性和存活并非必需。

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