Wang Yun, Kuo Hsien-Chi, Kuang Xiuli, Yao Shenqin, Lesnar Phil, Ng Lydia, Li Yaoyao, El-Hifnawi Laila, Chen Natalie, Zhang Kasey, Li Eric M, Ben-Simon Yoav, Ding Songlin, Wang Quanxin, Karlsson Totte, Delley Rachel, Willams Grace, Xiong Wei, Chen Chao, Chen Kai, Huang Zili, Yu Zhi-Feng, Xu Wenjie, Ahmadinia Leila, Walling-Bell Sarah, Andrade Julia, Gliko Olga, Sümbül Uygar, Mallory Matt, Farrel Colin, Sutton Ben, Jin Kelly, Yao Zizhen, Esposito Luke, Sunkin Susan, Kruse Lauren, Luo Qingming, Gong Hui, Li Anan, Qu Jia, Choi Hannah, Mihalas Stefan, Zhuang Jun, Zeng Hongkui, Sorensen Staci
Allen Institute for Brain Science, Seattle, Washington, USA.
State Key Laboratory of Eye Health, Eye Hospital, Wenzhou Medical University, Wenzhou, China.
bioRxiv. 2025 May 8:2025.05.08.652699. doi: 10.1101/2025.05.08.652699.
Molecularly defined cortical cell types have recently been linked to whole neuronal morphology (WNM), particularly those characterized by whole-brain-wide projections, such as intratelencephalic (IT), extratelencephalic (ET), and corticothalamic (CT) neurons. In contrast, classical morphological classifications (e.g., tufted TPC, small tufted SPC, and stellate SSC) are based primarily on local dendrosomatic and axonal structures, especially apical dendrites. This study bridges these perspectives by establishing a new neuronal taxonomy, analyzing the connectomes of defined cortical cell types, and comparing them with those obtained from bulk anterograde injections. Neurons were sparsely labeled via tamoxifen-inducible Cre lines with GFP reporters, and 1,419 WNM cells were comprehensively reconstructed with Vaa3D-TeraVR from ~15 areas across six functional regions of molecularly labeled brains imaged with 2p-fMOST. These cells were newly classified by integrating current molecular-WNM and classical morphological perspectives, with sample size augmented by 1,455 publicly available WNM cells reconstructed from the Mouse-Light project and CEBSIT. This effort defined ten combined molecular-WNM-classical morphological cell types: L5ET_TPC, L6CT_NPC, L6b_HPC, and seven IT types-L2/3IT_TPC, L4IT_SSC, L4IT_UPC, L4IT_TPC, L5IT_SPC, L6IT_IPC, and L6IT_car3PC. Clustering, quantitative analyses and random Forest classifier objectively validated these types and revealed their distinct connectomes, along with convergent, topographic, and hierarchical organizations across their projection brain regions. At the single-cell level, multiple organizational principles governing cortico-cortical (C-C) and cortico-subcortical (C-subC) connectomes emerged with unprecedented detail, offering a precise GPS-like tool for recordings and robust datasets for neuronal network modeling. Comparisons with bulk anterograde injection data underscored the limitations of traditional methods in identifying projection targets. Overall, our approach provides significant insights into cortical circuitry and elucidates the complex interplay between neuronal molecular identity, whole morphology, and classical morphological classification.
分子定义的皮质细胞类型最近已与整个神经元形态(WNM)相关联,特别是那些以全脑范围投射为特征的细胞类型,如脑内(IT)、脑外(ET)和皮质丘脑(CT)神经元。相比之下,经典的形态学分类(例如,簇状TPC、小簇状SPC和星状SSC)主要基于局部树突体和轴突结构,尤其是顶端树突。本研究通过建立一种新的神经元分类法、分析已定义皮质细胞类型的连接组,并将它们与从大量顺行注射获得的连接组进行比较,从而弥合了这些观点。通过带有GFP报告基因的他莫昔芬诱导型Cre系对神经元进行稀疏标记,并使用Vaa3D-TeraVR从用2p-fMOST成像的分子标记大脑的六个功能区域的约15个区域全面重建了1419个WNM细胞。通过整合当前的分子-WNM和经典形态学观点对这些细胞进行了新的分类,样本量通过从Mouse-Light项目和CEBSIT重建的1455个公开可用的WNM细胞得到了增加。这项工作定义了十种组合的分子-WNM-经典形态学细胞类型:L5ET_TPC、L6CT_NPC、L6b_HPC,以及七种IT类型-L2/3IT_TPC、L4IT_SSC、L4IT_UPC、L4IT_TPC、L5IT_SPC、L6IT_IPC和L6IT_car3PC。聚类、定量分析和随机森林分类器客观地验证了这些类型,并揭示了它们独特的连接组,以及它们投射脑区的汇聚、地形和层次组织。在单细胞水平上,支配皮质-皮质(C-C)和皮质-皮质下(C-subC)连接组的多种组织原则以前所未有的细节出现,为记录提供了一种精确的类似GPS的工具,并为神经元网络建模提供了强大的数据集。与大量顺行注射数据的比较突出了传统方法在识别投射靶点方面的局限性。总体而言,我们的方法为皮质回路提供了重要见解,并阐明了神经元分子身份、整体形态和经典形态学分类之间的复杂相互作用。
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