Leibniz Institute for Age Research - Fritz Lipmann Institute (FLI), Molecular Genetics, Jena, Germany.
Dev Dyn. 2012 Jun;241(6):1133-41. doi: 10.1002/dvdy.23789. Epub 2012 Apr 26.
A challenge in age research is the absence of short-lived vertebrate model organisms. The turquoise killifish Nothobranchius furzeri has an exceptionally short lifespan of 4-10 months depending on the strain. Thus, it possesses the shortest known maximum lifespan of a vertebrate species that can be bred in captivity.
Here we show the successful introduction of DNA and RNA molecules into the one-cell embryo of N. furzeri. For this purpose, we adapted existing microinjection protocols to inject through the remarkably thick and robust chorion of N. furzeri's eggs. The injected DNA transgene was integrated into the genome and transmitted to subsequent generations as indicated by the expression of the fluorophore enhanced green fluorescent protein (EGFP). Furthermore, we could confirm a special phase during embryonic development in which embryogenesis occurs within a re-aggregated mass of previously dispersed cells as it has been described for other related cyprinodont fish species.
The transgenesis protocol described here provides a basis for a variety of genetic manipulations including overexpression of genes and determining their effects on lifespan and longevity. The feasibility to perform transgenesis is an important step to establish N. furzeri as a new model in age research.
在衰老研究中面临的一个挑战是缺乏寿命较短的脊椎动物模式生物。翠绿色的半口脂鲤（Nothobranchius furzeri）的寿命极短，取决于品系，其寿命为 4-10 个月。因此，它具有已知的脊椎动物物种中最短的最大寿命，并且可以在圈养中繁殖。
在这里，我们展示了将 DNA 和 RNA 分子成功引入半口脂鲤的单细胞胚胎中的过程。为此，我们对现有的显微注射方案进行了改进，以便通过半口脂鲤的卵的显著厚而坚固的卵壳进行注射。注射的 DNA 转基因整合到基因组中，并通过荧光蛋白增强型绿色荧光蛋白（EGFP）的表达传递到后代，表明了这一点。此外，我们可以确认胚胎发育过程中的一个特殊阶段，其中胚胎发生发生在先前分散的细胞重新聚集的团块内，这与其他相关的脂鲤科鱼类物种的描述一致。
本文描述的转基因技术为各种基因操作提供了基础，包括基因的过表达以及确定它们对寿命和长寿的影响。进行转基因的可行性是将半口脂鲤确立为衰老研究中新模型的重要步骤。
