Dense functional and molecular readout of a circuit hub in sensory cortex.
Dense functional and molecular readout of a circuit hub in sensory cortex.
复制标题
感觉皮层回路中枢的密集功能和分子读出。
DOI：
10.1126/science.abl5981
复制
发表时间：
2022-01-07
期刊：
影响因子：
--
通讯作者：
中科院分区：
文献类型：
--
作者：
细胞类型的多样性是构成哺乳动物皮层的区域和层的神经元回路的定义特征。在分子水平上，这种多样性的程度，现在更好地理解，通过最近的努力，普查所有潜在的皮质细胞类型，通过单细胞转录谱。皮质群体可以分层细分为多个假定的转录组细胞类，亚类和类型。这种新的神经元亚类和亚型目录开辟了新的问题和研究途径，即这些细胞类型如何共同组织成电路，以处理信息并适应经验的变化。我们研究了初级体感皮层第2层或第3层（L2/3）中新发现的细胞类型的功能，该区域整合了自下而上的感觉信息和自上而下的内部表征。目前的体内方法主要允许一次研究一种细胞类型，并且标记由表达基因的组合限定的细胞类型的能力有限。为了密集地调查这些细胞类型并研究它们在任务行为期间如何相互作用，我们开发了一个平台，活性和细胞类型标记物的综合读数（CRACK），该平台将群体钙成像与随后的多重荧光原位杂交相结合。mRNA转录本的多重标记对于破译由基因表达的组合模式定义的细胞类型的身份至关重要。我们分析了三种兴奋性细胞类型和八个抑制性亚类在L2/3小鼠进行了基于胡须的触觉工作记忆任务的功能反应。兴奋性和抑制性神经元的任务相关特性随着它们被分离成越来越离散的分子类型而继续分化。我们的分析表明，兴奋性细胞类型，L2/3端脑内Baz 1a（Baz 1a），作为一个高度活跃的触觉功能检测器。同时成像识别的细胞类型，使亚群之间的功能连接的测量。功能连接分析表明，Baz 1a神经元协调局部网络活动模式。我们发现，Baz 1a神经元与树突靶向的、表达生长抑素（Sst）的抑制性神经元表现出强的功能联系。跨单突触病毒示踪证实，Baz 1a神经元优先突触到Sst神经元。Baz 1a神经元也显示出选择可塑性相关的立即早期基因的富集，包括Fos。为了确定立即早期基因的表达模式是否是Baz 1a神经元的稳定特性以及这与神经元可塑性的关系，我们跟踪了胡须剥夺小鼠的Fos表达和神经元活动。我们发现，Baz 1a神经元稳态适应感觉剥夺，同时稳定维持Fos表达。这些结果表明，Baz 1a神经元是一个分子定义的电路基序的组成部分，能够招募局部电路的感觉处理时，在行为过程中遇到的显着特征。这种细胞类型还可以在持续和改变的感觉体验中保持感觉表征。这是基于我们对躯体感觉皮层中的局部回路如何实现自下而上和自上而下信息协商的知识。绘制神经元群体之间的功能和转录关系的能力，提供了对皮层的组织原则如何产生其执行的计算的洞察。CRACK平台揭示了感官处理的电路中心。功能分析的分子定义的细胞，实现了在体内的L2/3的初级体感皮层在任务行为或感觉剥夺，然后多重荧光原位杂交的双光子钙成像。兴奋性Baz 1a神经元形成连接基序，能够在持续和改变的感觉体验期间招募局部回路并保留感觉表征。HCR-FISH，杂交链反应-荧光原位杂交; Vip，血管活性肠肽表达; Exc.，兴奋虽然新皮层的单细胞转录组学已经发现了300多种假定的细胞类型，但这种分子分类是否预测了不同的功能作用尚不清楚。我们将双光子钙成像与空间转录组学相结合，从功能和分子水平研究皮质回路。我们的特点是行为相关的反应，在第2层或第3层的初级躯体感觉皮层的主要神经元亚类小鼠进行触觉工作记忆任务。我们确定了一种兴奋性端脑内细胞类型，Baz 1a，表现出高触觉功能的选择性。Baz 1a神经元在改变的经历中保持对刺激的反应性，并显示出立即早期基因子集的持续富集。功能和解剖学的连接表明，Baz 1a神经元居住在第2层或第3层的上部优先支配生长抑素表达抑制神经元。这个基序定义了一个回路中枢，协调新皮层表层的局部感觉处理。
The diversity of cell types is a defining feature of the neuronal circuitry that makes up the areas and layers of the mammalian cortex. At a molecular level, the extent of this diversity is now better appreciated through recent efforts to census all potential cortical cell types through single-cell transcriptional profiling. Cortical populations can be hierarchically subdivided into multiple putative transcriptomic cell classes, subclasses, and types. This new catalog of neuronal subclasses and subtypes opens up new questions and avenues of investigation for how these cell types are collectively organized into circuits that function to process information and adapt to changes in experience. We investigated the function of newly identified cell types in layers 2 or 3 (L2/3) of the primary somatosensory cortex, a region that integrates bottom-up sensory information with top-down internal representations. Current in vivo methods primarily allow cell types to be investigated one at a time and have limited ability to label cell types defined by combinations of expressed genes. To densely survey these cell types and investigate how they interact during task behavior, we developed a platform, Comprehensive Readout of Activity and Cell Type Markers (CRACK), that combines population calcium imaging with subsequent multiplexed fluorescent in situ hybridization. Multiplexed labeling of mRNA transcripts is critical to deciphering the identity of cell types defined by combinatorial patterns of gene expression. We profiled the functional responses of three excitatory cell types and eight inhibitory subclasses in L2/3 as mice performed a whisker-based tactile working memory task. Task-related properties of both excitatory and inhibitory neurons continue to differentiate as they are segregated into increasingly discrete molecular types. Our analysis revealed that the excitatory cell type, L2/3 intratelencephalic Baz1a (Baz1a), functions as a highly active detector of tactile features. Simultaneous imaging across identified cell types enabled measurements of functional connectivity between subpopulations. Functional connectivity analysis indicated that Baz1a neurons orchestrate local network activity patterns. We found that Baz1a neurons show strong functional connections with dendrite-targeting, somatostatin-expressing (Sst) inhibitory neurons. Trans-monosynaptic viral tracing confirmed that Baz1a neurons preferentially synapse onto Sst neurons. Baz1a neurons also show enrichment of select plasticity-related, immediate early genes, including Fos. To determine whether the expression pattern of immediate early genes is a stable property of Baz1a neurons and how this relates to neuronal plasticity, we tracked Fos expression and neuronal activity in mice subjected to whisker deprivation. We found that Baz1a neurons homeostatically adapt to sensory deprivation while stably maintaining Fos expression. These results demonstrate that Baz1a neurons are a component of a molecularly defined circuit motif that is capable of recruiting local circuits for sensory processing when salient features are encountered during behavior. This cell type also functions to preserve sensory representations during ongoing and altered sensory experience. This builds on our knowledge for how local circuits in somatosensory cortex are implemented to negotiate bottom-up and top-down information. The ability to map functional and transcriptional relationships across neuronal populations provides insight into how the organizing principles of the cortex give rise to the computations it performs. CRACK platform reveals a circuit hub for sensory processing. Functional profiling of molecularly defined cells was achieved with in vivo two-photon calcium imaging in L2/3 of the primary somatosensory cortex during task behavior or sensory deprivation followed by multiplexed fluorescent in situ hybridization. Excitatory Baz1a neurons form a connection motif capable of recruiting local circuits and preserving sensory representations during ongoing and altered sensory experience. HCR-FISH, hybridization chain reaction–fluorescence in situ hybridization; Vip, vasoactive intestinal peptide–expressing; Exc., excitatory. Although single-cell transcriptomics of the neocortex has uncovered more than 300 putative cell types, whether this molecular classification predicts distinct functional roles is unclear. We combined two-photon calcium imaging with spatial transcriptomics to functionally and molecularly investigate cortical circuits. We characterized behavior-related responses across major neuronal subclasses in layers 2 or 3 of the primary somatosensory cortex as mice performed a tactile working memory task. We identified an excitatory intratelencephalic cell type, Baz1a, that exhibits high tactile feature selectivity. Baz1a neurons homeostatically maintain stimulus responsiveness during altered experience and show persistent enrichment of subsets of immediately early genes. Functional and anatomical connectivity reveals that Baz1a neurons residing in upper portions of layers 2 or 3 preferentially innervate somatostatin-expressing inhibitory neurons. This motif defines a circuit hub that orchestrates local sensory processing in superficial layers of the neocortex.
登录
查看更多内容
影响因子：
16.2
作者：
Hill DN;Curtis JC;Moore JD;Kleinfeld D
通讯作者：
Kleinfeld D
小鼠大脑皮层 GABA 神经元组合靶向的策略和工具
影响因子：
16.2
作者：
He M;Tucciarone J;Lee S;Nigro MJ;Kim Y;Levine JM;Kelly SM;Krugikov I;Wu P;Chen Y;Gong L;Hou Y;Osten P;Rudy B;Huang ZJ
通讯作者：
Huang ZJ
影响因子：
25
作者：
Chan KY;Jang MJ;Yoo BB;Greenbaum A;Ravi N;Wu WL;Sánchez-Guardado L;Lois C;Mazmanian SK;Deverman BE;Gradinaru V
通讯作者：
Gradinaru V
影响因子：
5.8
作者：
Arganda-Carreras, Ignacio;Kaynig, Verena;Seung, H. Sebastian
通讯作者：
Seung, H. Sebastian
影响因子：
5.3
作者：
Barth, AL;Gerkin, RC;Dean, KL
通讯作者：
Dean, KL