2P-NucTag: on-demand phototagging for molecular analysis of functionally identified cortical neurons.
2P-NucTag: on-demand phototagging for molecular analysis of functionally identified cortical neurons.
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2P-NucTag：按需照片标记，用于功能识别的皮质神经元的分子分析。
DOI：
10.1101/2024.03.21.586118
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发表时间：
2024
期刊：
影响因子：
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通讯作者：
Spiegel,Ivo
中科院分区：
文献类型：
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作者：
Shi,Jingcheng;Nutkovich,Boaz;Kushinsky,Dahlia;Rao,BoveyY;Herrlinger,StephanieA;Mihaila,TiberiuS;Malina,KatayunCohen-Kashi;O'Toole,CliodhnaK;CondeParedes,MargaretE;Yong,HyunChoong;Varol,Erdem;Losonczy,Attila;Spiegel,Ivo
神经回路的特点是基因和功能不同的细胞类型。对电路功能的机械理解是建立在将单个神经元的遗传和生理特性联系起来的基础上的。然而，将转录特性映射到哺乳动物体内皮层回路中功能异质的神经元亚型仍然具有高度挑战性。在这里，我们介绍了一种高通量双光子核光标记（p - nuctag）方法，该方法通过光激活红色荧光蛋白在行为小鼠体内功能表征后，优化了单个神经元的按需和不可磨灭标记。我们通过选择性地标记和转录分析小鼠海马中以前无法获得的“位置”和“沉默”细胞，证明了这种功能向前管道的实用性。我们的研究结果揭示了这些具有不同空间编码特性的海马锥体神经元之间意想不到的基因表达差异。因此，2P-NucTag为揭示控制神经回路功能组织的分子原理开辟了一条新途径。
Neural circuits are characterized by genetically and functionally diverse cell types. A mechanistic understanding of circuit function is predicated on linking the genetic and physiological properties of individual neurons. However, it remains highly challenging to map the transcriptional properties to functionally heterogeneous neuronal subtypes in mammalian cortical circuits in vivo. Here, we introduce a high-throughput two-photon nuclear phototagging (2P-NucTag) approach optimized for on-demand and indelible labeling of single neurons via a photoactivatable red fluorescent protein following in vivo functional characterization in behaving mice. We demonstrate the utility of this function-forward pipeline by selectively labeling and transcriptionally profiling previously inaccessible ‘place’ and ‘silent’ cells in the mouse hippocampus. Our results reveal unexpected differences in gene expression between these hippocampal pyramidal neurons with distinct spatial coding properties. Thus, 2P-NucTag opens a new way to uncover the molecular principles that govern the functional organization of neural circuits.