Berlin Shai, Carroll Elizabeth C, Newman Zachary L, Okada Hitomi O, Quinn Carson M, Kallman Benjamin, Rockwell Nathan C, Martin Shelley S, Lagarias J Clark, Isacoff Ehud Y
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA.
Helen Wills Neuroscience Institute, University of California, Berkeley, Berkeley, California, USA.
Nat Methods. 2015 Sep;12(9):852-8. doi: 10.1038/nmeth.3480. Epub 2015 Jul 13.
Circuit mapping requires knowledge of both structural and functional connectivity between cells. Although optical tools have been made to assess either the morphology and projections of neurons or their activity and functional connections, few probes integrate this information. We have generated a family of photoactivatable genetically encoded Ca(2+) indicators that combines attributes of high-contrast photolabeling with high-sensitivity Ca(2+) detection in a single-color protein sensor. We demonstrated in cultured neurons and in fruit fly and zebrafish larvae how single cells could be selected out of dense populations for visualization of morphology and high signal-to-noise measurements of activity, synaptic transmission and connectivity. Our design strategy is transferrable to other sensors based on circularly permutated GFP (cpGFP).
回路测绘需要了解细胞之间的结构和功能连接。尽管已经有光学工具用于评估神经元的形态和投射或者它们的活性及功能连接，但很少有探针能整合这些信息。我们已经生成了一系列光激活的基因编码钙指示剂，它们在单一颜色的蛋白质传感器中结合了高对比度光标记和高灵敏度钙检测的特性。我们在培养的神经元、果蝇和斑马鱼幼虫中证明了如何从密集群体中挑选出单个细胞，以可视化其形态并对活性、突触传递和连接性进行高信噪比测量。我们的设计策略可转移到基于环状排列绿色荧光蛋白（cpGFP）的其他传感器上。
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