A three-dimensional single-cell-resolution whole-brain atlas using CUBIC-X expansion microscopy and tissue clearing
A three-dimensional single-cell-resolution whole-brain atlas using CUBIC-X expansion microscopy and tissue clearing
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DOI：
10.1038/s41593-018-0109-1
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发表时间：
2018-04-01
影响因子：
25
通讯作者：
Ueda, Hiroki R.
中科院分区：
文献类型：
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作者：
Murakami, Tatsuya C.;Mano, Tomoyuki;Ueda, Hiroki R.
三维单细胞分辨率的哺乳动物脑图谱将加速系统级识别和分析各种脑功能背后的细胞回路。然而，它的构建需要贯穿整个大脑的高效亚细胞分辨率成像。为了解决这一挑战，我们开发了一种基于水化学溶液（cbic - x）的荧光蛋白兼容、全器官清除和均匀扩增方案。扩展的、清晰的大脑使我们能够构建一个基于点的单细胞注释小鼠脑图谱（cbic - atlas）。cbic - atlas反映了全脑发育的不均匀性，揭示了出生后发育过程中大脑视觉和体感觉皮质区域的显著减少。对药物刺激的Arc-dVenus报告小鼠大脑进行概率活动映射到cbic - atlas上，发现海马齿状回存在不同的功能结构。cbic - atlas可通过一个基于web的开源查看器共享，为全脑细胞分析提供了一个新的平台。
A three-dimensional single-cell-resolution mammalian brain atlas will accelerate systems-level identification and analysis of cellular circuits underlying various brain functions. However, its construction requires efficient subcellular-resolution imaging throughout the entire brain. To address this challenge, we developed a fluorescent-protein-compatible, whole-organ clearing and homogeneous expansion protocol based on an aqueous chemical solution (CUBIC-X). The expanded, well-cleared brain enabled us to construct a point-based mouse brain atlas with single-cell annotation (CUBIC-Atlas). CUBIC-Atlas reflects inhomogeneous whole-brain development, revealing a significant decrease in the cerebral visual and somatosensory cortical areas during postnatal development. Probabilistic activity mapping of pharmacologically stimulated Arc-dVenus reporter mouse brains onto CUBIC-Atlas revealed the existence of distinct functional structures in the hippocampal dentate gyrus. CUBIC-Atlas is shareable by an open-source web-based viewer, providing a new platform for whole-brain cell profiling.