Simultaneous visualization of multiple neuronal properties with single-cell resolution in the living rodent brain
Simultaneous visualization of multiple neuronal properties with single-cell resolution in the living rodent brain
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DOI：
10.1016/j.mcn.2011.08.005
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发表时间：
2011-11-01
影响因子：
3.5
通讯作者：
Kawasaki, Hiroshi
中科院分区：
文献类型：
--
作者：
Ako, Rie;Wakimoto, Mayu;Kawasaki, Hiroshi
为了了解体内单个神经元的精细结构和功能特性，我们开发并验证了一种快速遗传技术，该技术能够同时研究活体啮齿动物大脑中具有单细胞分辨率的多个神经元特性。我们的技术PASME（启动子辅助稀疏神经元多基因标记使用在子宫内电穿孔）的目标是特定的小子集的稀疏锥体神经元在第2/3层，第5层的大脑皮层和海马与多个荧光报告蛋白，如突触后PSD-95-GFP和GFP-桥蛋白。该技术也适用于独立地靶向单个神经元及其来自周围神经元的突触前输入。针对稀疏的第2/3层神经元，我们发现了小鼠大脑皮层中的第2/3层神经元的新亚群。这种技术，广泛适用于探测和操纵神经元与单细胞分辨率在体内，应该提供一个强大的手段来揭示大脑所采用的基本机制，特别是当结合在体内双光子激光扫描显微镜和/或光遗传学技术。(C)2011 Elsevier Inc.版权所有© 2016
To understand the fine-scale structures and functional properties of individual neurons in vivo, we developed and validated a rapid genetic technique that enables simultaneous investigation of multiple neuronal properties with single-cell resolution in the living rodent brain. Our technique PASME (promoter-assisted sparse-neuron multiple-gene labeling using in utero electroporation) targets specific small subsets of sparse pyramidal neurons in layer 2/3, layer 5 of the cerebral cortex and in the hippocampus with multiple fluorescent reporter proteins such as postsynaptic PSD-95-GFP and GFP-gephyrin. The technique is also applicable for targeting independently individual neurons and their presynaptic inputs derived from surrounding neurons. Targeting sparse layer 2/3 neurons, we uncovered a novel subpopulation of layer 2/3 neurons in the mouse cerebral cortex. This technique, broadly applicable for probing and manipulating neurons with single-cell resolution in vivo, should provide a robust means to uncover the basic mechanisms employed by the brain, especially when combined with in vivo two-photon laser-scanning microscopy and/or optogenetic technologies. (C) 2011 Elsevier Inc. All rights reserved,