Photoactivatable genetically encoded calcium indicators for targeted neuronal imaging.
Photoactivatable genetically encoded calcium indicators for targeted neuronal imaging.
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DOI：
10.1038/nmeth.3480
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发表时间：
2015-09
期刊：
影响因子：
48
通讯作者：
Isacoff EY
中科院分区：
文献类型：
--
作者：
Berlin S;Carroll EC;Newman ZL;Okada HO;Quinn CM;Kallman B;Rockwell NC;Martin SS;Lagarias JC;Isacoff EY
电路映射需要了解细胞之间的结构和功能连接。虽然光学工具已被用于评估神经元的形态和投射或其活动和功能连接，但很少有探针整合这些信息。我们已经产生了一个家庭的光活化的遗传编码的Ca 2+指标（pa-GECIs），结合高对比度的光标记与高灵敏度的Ca 2+检测在一个单一的颜色，蛋白质传感器的属性。我们证明了在培养的神经元和在体内的果蝇和斑马鱼幼虫的pa-GECIs的效用。我们展示了如何从密集的群体中选择单个细胞，以进行高尔基体样的形态可视化和高信噪比的活动，突触传递和连接测量。我们的设计策略是很容易转移到其他传感器的基础上循环排列的GFP（cpGFP）。
Circuit mapping requires knowledge of both structural and functional connectivity between cells. While 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 Ca2+ Indicators (pa-GECIs) that combines attributes of high-contrast photo-labeling with high-sensitivity Ca2+ detection in a single-color, protein-sensor. We demonstrate the utility of pa-GECIs in cultured neurons and in vivo in Drosophila and zebrafish larvae. We show how single cells can be selected out of dense populations for Golgi-like visualization of morphology and high signal-to-noise measurements of activity, synaptic transmission and connectivity. Our design strategy is readily transferrable to other sensors based on circularly permutated GFP (cpGFP).