A germline GFP transgenic axolotl and its use to track cell fate: Dual origin of the fin mesenchyme during development and the fate of blood cells during regeneration
A germline GFP transgenic axolotl and its use to track cell fate: Dual origin of the fin mesenchyme during development and the fate of blood cells during regeneration
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DOI：
10.1016/j.ydbio.2005.11.037
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发表时间：
2006-02-15
影响因子：
2.7
通讯作者：
Tanaka, EM
中科院分区：
文献类型：
--
作者：
Sobkow, L;Epperlein, HH;Tanaka, EM
在蝾螈中转基因的发展对于研究发育和再生至关重要，因为它将允许长期的细胞命运追踪以及基因表达分析。我们在这里证明，质粒注射到一个细胞阶段的蝾螈胚胎产生镶嵌转基因动物，显示生殖系传输的转基因。在注射液中包含SceI大范围核酸酶（Thermes，V，Grabher，C.，Ristoratore，F.，Bourrat，F.，Choulika，A.，Wittbrodt，J.，Joly，J.S.，2002. I-SceI大范围核酸酶介导鱼类高效转基因机械设备118，91 - 98）导致更高百分比的FO动物在全身显示强表达。这代表了转基因的种系传递在蝾螈中的第一个证明。使用这种技术，我们已经产生了一个生殖系转基因动物表达GFP无处不在的所有组织检查。我们用这种动物来研究背鳍在发育过程中的细胞命运。我们已经发现的体节细胞的背鳍间充质的蝾螈，这是以前假定仅来自神经嵴的贡献。我们还研究了血液在尾再生过程中的作用，通过将来自GFP（+）胚胎的腹侧血液形成区移植到未标记的宿主中。在尾部再生过程中，我们没有观察到GFP+细胞对肌肉或神经有贡献，这表明在尾部再生过程中造血干细胞没有经历显著的可塑性。(C)2005年爱思唯尔公司All rights reserved.
The development of transgenesis in axolotls is crucial for studying development and regeneration as it would allow for long-term cell fate tracing as well as gene expression analysis. We demonstrate here that plasmid injection into the one-cell stage axolotl embryo generates mosaic transgenic animals that display germline transmission of the transgene. The inclusion of SceI meganuclease in the injections (Thermes, V, Grabher, C., Ristoratore, F., Bourrat, F., Choulika, A., Wittbrodt, J., Joly, J.S., 2002. I-SceI meganuclease mediates highly efficient transgenesis in fish. Mech. Dev. 118, 91 -98) resulted in a higher percentage of F0 animals displaying strong expression throughout the body. This represents the first demonstration in the axolotl of germline transmission of a transgene. Using this technique we have generated a germline transgenic animal expressing GFP ubiquitously in all tissues examined. We have used this animal to study cell fate in the dorsal fin during development. We have uncovered a contribution of somite cells to dorsal fin mesenchyme in the axolotl, which was previously assumed to derive solely from neural crest. We have also studied the role of blood during tail regeneration by transplanting the ventral blood-forming region from GFP(+) embryos into unlabeled hosts. During tail regeneration, we do not observe GFP+ cells contributing to muscle or nerve, suggesting that during tail regeneration blood stem cells do not undergo significant plasticity. (C) 2005 Elsevier Inc. All rights reserved.