CRASP：跨突触伙伴的CFP重组
CRASP: CFP reconstitution across synaptic partners.
作者信息
Li Yiming, Guo Aike, Li Hao
机构信息
Institute of Neuroscience, State Key Laboratory of Neuroscience, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China; University of Chinese Academy of Sciences, Beijing 100049, China.
Institute of Neuroscience, State Key Laboratory of Neuroscience, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China; State Key Laboratory of Brain and Cognitive Science, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China.
出版信息
Biochem Biophys Res Commun. 2016 Jan 15;469(3):352-6. doi: 10.1016/j.bbrc.2015.12.011. Epub 2015 Dec 10.
Mapping the pattern of connectivity between neurons is widely regarded to be critical for understanding the nervous system. GRASP (GFP reconstitution across synaptic partners) has been used as a promising method for mapping neuronal connectivity, but is currently available in the green color only, limiting its potential applications. Here we demonstrate CRASP (CFP reconstitution across synaptic partners), a cyan-colored version of GRASP. We validated the system in HEK 293T cells, and generated transgenic Drosophila lines to show that the system could reliably detect neuronal contacts in the brain. Furthermore, we showed that the CRASP signal could be selectively amplified using standard immunohistochemistry methods. The CRASP system adds to the toolkit available to researchers for mapping neuronal connectivity, and substantially expands the potential application of GRASP-like strategies.
绘制神经元之间的连接模式被广泛认为对于理解神经系统至关重要。GRASP（跨突触伙伴的绿色荧光蛋白重组）已被用作绘制神经元连接的一种有前景的方法，但目前仅有一种绿色版本，这限制了其潜在应用。在此，我们展示了CRASP（跨突触伙伴的青色荧光蛋白重组），它是GRASP的青色版本。我们在HEK 293T细胞中验证了该系统，并生成了转基因果蝇品系，以表明该系统能够可靠地检测大脑中的神经元接触。此外，我们表明可以使用标准免疫组织化学方法选择性地放大CRASP信号。CRASP系统为研究人员绘制神经元连接增添了可用工具，并大幅扩展了类似GRASP策略的潜在应用。
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