Patiño Maribel, Rossa Marley A, Lagos Willian Nuñez, Patne Neelakshi S, Callaway Edward M
Systems Neurobiology Laboratory, The Salk Institute for Biological Studies, La Jolla, CA, USA; Medical Scientist Training Program, University of California, San Diego, La Jolla, CA, USA.
Systems Neurobiology Laboratory, The Salk Institute for Biological Studies, La Jolla, CA, USA; Neurosciences Graduate Program, University of California, San Diego, La Jolla, CA, USA.
Neuron. 2024 Dec 4;112(23):3851-3866.e4. doi: 10.1016/j.neuron.2024.09.003. Epub 2024 Sep 30.
Complex neocortical functions rely on networks of diverse excitatory and inhibitory neurons. While local connectivity rules between major neuronal subclasses have been established, the specificity of connections at the level of transcriptomic subtypes remains unclear. We introduce single transcriptome assisted rabies tracing (START), a method combining monosynaptic rabies tracing and single-nuclei RNA sequencing to identify transcriptomic cell types, providing inputs to defined neuron populations. We employ START to transcriptomically characterize inhibitory neurons providing monosynaptic input to 5 different layer-specific excitatory cortical neuron populations in mouse primary visual cortex (V1). At the subclass level, we observe results consistent with findings from prior studies that resolve neuronal subclasses using antibody staining, transgenic mouse lines, and morphological reconstruction. With improved neuronal subtype granularity achieved with START, we demonstrate transcriptomic subtype specificity of inhibitory inputs to various excitatory neuron subclasses. These results establish local connectivity rules at the resolution of transcriptomic inhibitory cell types.
复杂的新皮层功能依赖于由多种兴奋性和抑制性神经元组成的网络。虽然主要神经元亚类之间的局部连接规则已经确立，但转录组亚型水平上连接的特异性仍不清楚。我们引入了单转录组辅助狂犬病示踪（START）方法，该方法结合了单突触狂犬病示踪和单细胞核RNA测序来识别转录组细胞类型，为特定的神经元群体提供输入。我们利用START对抑制性神经元进行转录组特征分析，这些抑制性神经元为小鼠初级视觉皮层（V1）中5种不同层特异性兴奋性皮层神经元群体提供单突触输入。在亚类水平上，我们观察到的结果与先前使用抗体染色、转基因小鼠系和形态重建来解析神经元亚类的研究结果一致。通过START实现了更高的神经元亚型分辨率，我们证明了对各种兴奋性神经元亚类的抑制性输入的转录组亚型特异性。这些结果在转录组抑制性细胞类型的分辨率上确立了局部连接规则。
