Gomez-Nicola Diego, Riecken Kristoffer, Fehse Boris, Perry V Hugh
Centre for Biological Sciences, University of Southampton, Southampton, United Kingdom.
Research Department Cell and Gene Therapy, Clinic for Stem Cell Transplantation, University Medical Centre Hamburg-Eppendorf, Hamburg, Germany.
Sci Rep. 2014 Dec 22;4:7520. doi: 10.1038/srep07520.
In neuroscience it is a technical challenge to identify and follow the temporal and spatial distribution of cells as they differentiate. We hypothesised that RGB marking, the tagging of individual cells with unique hues resulting from simultaneous expression of the three basic colours red, green and blue, provides a convenient toolbox for the study of the CNS anatomy at the single-cell level. Using γ-retroviral and lentiviral vector sets we describe for the first time the in-vivo multicolour RGB marking of neurons in the adult brain. RGB marking also enabled us to track the spatial and temporal fate of neural stem cells in the adult brain. The application of different viral envelopes and promoters provided a useful approach to track the generation of neurons vs. glial cells at the neurogenic niche, allowing the identification of the prominent generation of new astrocytes to the striatum. Multicolour RGB marking could serve as a universal and reproducible method to study and manipulate the CNS at the single-cell level, in both health and disease.
在神经科学领域，识别并追踪细胞分化过程中的时空分布是一项技术挑战。我们推测，RGB标记法，即通过同时表达红、绿、蓝三种基本颜色，用独特色调对单个细胞进行标记，为在单细胞水平研究中枢神经系统解剖结构提供了一个便利的工具包。我们首次使用γ-逆转录病毒和慢病毒载体组描述了成体大脑中神经元的体内多色RGB标记。RGB标记还使我们能够追踪成体大脑中神经干细胞的时空命运。应用不同的病毒包膜和启动子为追踪神经发生微环境中神经元与神经胶质细胞的生成提供了一种有用的方法，从而能够识别出纹状体中显著产生的新星形胶质细胞。多色RGB标记可作为一种通用且可重复的方法，用于在健康和疾病状态下的单细胞水平研究和操纵中枢神经系统。
