Brain-wide 3D imaging of neuronal activity in Caenorhabditis elegans with sculpted light
Brain-wide 3D imaging of neuronal activity in Caenorhabditis elegans with sculpted light
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DOI：
10.1038/nmeth.2637
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发表时间：
2013-10-01
期刊：
影响因子：
48
通讯作者：
Vaziri, Alipasha
中科院分区：
文献类型：
--
作者：
Schrodel, Tina;Prevedel, Robert;Vaziri, Alipasha
最近神经科学研究的目标是获得详细的解剖神经元接线图，以及这些网络中的神经元如何参与动态活动的信息。尽管秀丽隐杆线虫神经系统的完整连接图谱已被发现超过25年，但这一知识还不足以预测所有潜在行为的功能连接。为了实现这一目标，我们开发了一种双光子技术，利用宽视场时间聚焦（WF-TeFo）对秀丽隐杆线虫进行全脑钙成像。对我们的结果至关重要的是使用核定位，遗传编码的钙指示剂NLS-GCaMP5K，它允许在秀丽隐杆线虫密集排列的头神经节内对单个神经元进行明确的区分。我们展示了几乎同时记录多达70%的头部神经元的活动。结合用于刺激传递的芯片实验室设备，该方法为建立神经元网络的功能图提供了一个有利的平台。
Recent efforts in neuroscience research have been aimed at obtaining detailed anatomical neuronal wiring maps as well as information on how neurons in these networks engage in dynamic activities. Although the entire connectivity map of the nervous system of Caenorhabditis elegans has been known for more than 25 years, this knowledge has not been sufficient to predict all functional connections underlying behavior. To approach this goal, we developed a two-photon technique for brain-wide calcium imaging in C. elegans, using wide-field temporal focusing (WF-TeFo). Pivotal to our results was the use of a nuclear-localized, genetically encoded calcium indicator, NLS-GCaMP5K, that permits unambiguous discrimination of individual neurons within the densely packed head ganglia of C. elegans. We demonstrate near-simultaneous recording of activity of up to 70% of all head neurons. In combination with a lab-on-a-chip device for stimulus delivery, this method provides an enabling platform for establishing functional maps of neuronal networks.