Charting Monosynaptic Connectivity Maps by Two-Color Light-Sheet Fluorescence Microscopy
Charting Monosynaptic Connectivity Maps by Two-Color Light-Sheet Fluorescence Microscopy
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DOI：
10.1016/j.celrep.2012.10.008
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发表时间：
2012-11-01
期刊：
影响因子：
8.8
通讯作者：
Schwarz, Martin K.
中科院分区：
文献类型：
--
作者：
Niedworok, Christian J.;Schwarz, Inna;Schwarz, Martin K.
未切割的成年小鼠大脑中定义的、荧光标记的长距离神经元网络的细胞分辨率三维 (3D) 可视化一直难以实现。在这里，描述了一种基于病毒的策略，该策略允许在单次立体定向注射到选定的神经元群体中后，对腹侧前脑中的离心投射神经元群体及其直接、单突触连接的延髓中间神经元进行荧光标记。改进的组织透明化与光片荧光显微镜相结合的实施允许在单次全脑扫描中对所得连接图进行成像。随后的 3D 重建揭示了与不同延髓中间神经元群单突触连接的不同神经元群的确切分布。此外，光片荧光成像后大脑的再水化能够对突触连接的神经元进行免疫组织化学鉴定。因此，这项研究描述了一种从不同的、病毒标记的神经元群体中识别单突触连接图的方法，有助于更好地理解神经系统中的信息流。
Cellular resolution three-dimensional (3D) visualization of defined, fluorescently labeled long-range neuronal networks in the uncut adult mouse brain has been elusive. Here, a virus-based strategy is described that allowed fluorescent labeling of centrifugally projecting neuronal populations in the ventral forebrain and their directly, monosynaptically connected bulbar interneurons upon a single stereotaxic injection into select neuronal populations. Implementation of improved tissue clearing combined with light-sheet fluorescence microscopy permitted imaging of the resulting connectivity maps in a single whole-brain scan. Subsequent 3D reconstructions revealed the exact distribution of the diverse neuronal ensembles monosynaptically connected with distinct bulbar interneuron populations. Moreover, rehydratation of brains after light-sheet fluorescence imaging enabled the immunohistochemical identification of synaptically connected neurons. Thus, this study describes a method for identifying monosynaptic connectivity maps from distinct, virally labeled neuronal populations that helps in better understanding of information flow in neural systems.