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体内高空间分辨率双光子双向控制和神经元兴奋性成像。

Two-Photon Bidirectional Control and Imaging of Neuronal Excitability with High Spatial Resolution In Vivo.

机构信息

Optical Approaches to Brain Function Laboratory, Istituto Italiano di Tecnologia, Genova 16163, Italy.

Department of Neurobiology, Weizmann Institute of Science, Rehovot 76100, Israel.

出版信息

Cell Rep. 2018 Mar 13;22(11):3087-3098. doi: 10.1016/j.celrep.2018.02.063.

DOI:10.1016/j.celrep.2018.02.063
PMID:29539433
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5863087/
Abstract

Sensory information is encoded within the brain in distributed spatiotemporal patterns of neuronal activity. Understanding how these patterns influence behavior requires a method to measure and to bidirectionally perturb with high spatial resolution the activity of the multiple neuronal cell types engaged in sensory processing. Here, we combined two-photon holography to stimulate neurons expressing blue light-sensitive opsins (ChR2 and GtACR2) with two-photon imaging of the red-shifted indicator jRCaMP1a in the mouse neocortex in vivo. We demonstrate efficient control of neural excitability across cell types and layers with holographic stimulation and improved spatial resolution by opsin somatic targeting. Moreover, we performed simultaneous two-photon imaging of jRCaMP1a and bidirectional two-photon manipulation of cellular activity with negligible effect of the imaging beam on opsin excitation. This all-optical approach represents a powerful tool to causally dissect how activity patterns in specified ensembles of neurons determine brain function and animal behavior.

摘要

感觉信息在大脑中以神经元活动的分布式时空模式进行编码。要了解这些模式如何影响行为，就需要一种方法来测量并以高空间分辨率双向扰动参与感觉处理的多个神经元细胞类型的活动。在这里，我们结合双光子全息术来刺激表达蓝光敏感光蛋白（ChR2 和 GtACR2）的神经元，同时在体内对红色位移指示剂 jRCaMP1a 进行双光子成像。我们证明了通过全息刺激可以在细胞类型和层之间进行有效的神经兴奋性控制，并通过光蛋白体细胞靶向实现了更高的空间分辨率。此外，我们还进行了 jRCaMP1a 的同时双光子成像和细胞活性的双向双光子操作，成像光束对光蛋白激发的影响可以忽略不计。这种全光学方法是一种强大的工具，可以用于因果剖析特定神经元集合的活动模式如何决定大脑功能和动物行为。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ee/5863087/4dd03eaafc56/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ee/5863087/b89907f4ddcf/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ee/5863087/d9d0b73137d2/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ee/5863087/38afd8883f6c/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ee/5863087/612ab59d0770/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ee/5863087/e0f9fa38c804/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ee/5863087/733c520f45de/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ee/5863087/4e38dde089d9/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ee/5863087/4dd03eaafc56/gr7.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ee/5863087/b89907f4ddcf/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ee/5863087/d9d0b73137d2/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ee/5863087/38afd8883f6c/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ee/5863087/612ab59d0770/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ee/5863087/e0f9fa38c804/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ee/5863087/733c520f45de/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ee/5863087/4e38dde089d9/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/97ee/5863087/4dd03eaafc56/gr7.jpg

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