Wiring and developmental principles of inhibitory neocortical circuits

抑制性新皮质回路的布线和发育原理

• 批准号: 10478363
• 负责人: HIROKI TANIGUCHI
• 金额: $ 35.44万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-01-21 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10478363
• 关键词: Address; Anatomy; Animal Disease Models; Area; Axon; Brain; Brain Diseases; Brain Pathology; Cells; Defect; Dependovirus; Development; Developmental Process; Diagnosis; Epilepsy; Equilibrium; Exhibits; Genetic; Interneurons; Knock-in Mouse; Knowledge; Label; Mediating; Methods; Morphology; Motor Cortex; Neocortex; Neurons; Parvalbumins; Physiological; Physiology; Positioning Attribute; Prevention strategy; Rabies virus; Regulation; Resolution; Role; Schizophrenia; Shapes; Somatosensory Cortex; Somatostatin; Structure; Techniques; Testing; Therapeutic; Time; Vasoactive Intestinal Peptide; Visual Cortex; Work; autism spectrum disorder; cell type; experimental study; genetic approach; genetic manipulation; in vivo; information processing; mouse genetics; neocortical; neurochemistry; neurotransmission; novel; novel strategies; postnatal; postsynaptic; recombinase; relating to nervous system; tool; transcription factor; treatment strategy

Project Summary/Abstract Inhibitory circuits formed by GABAergic interneurons (INs) contribute to processing and encoding of cortical information by shaping the spatial and temporal structure of neural activity. Consistent with this critical role of INs in normal brain functions, IN malfunction has been implicated in a wide array of brain disorders such as schizophrenia, autism, and epilepsy. Despite their importance, detailed wiring diagrams of inhibitory local circuits remain largely unknown due to huge diversity of IN types. Furthermore, it is poorly understood what principles govern assembly of inhibitory microcircuits. Filling these knowledge gaps will provide us with wiring and developmental principles of cortical inhibitory circuits, which in turn dramatically facilitate our understanding of how cortical circuits work. One fundamental cortical circuit module contains an excitatory principal neuron (PN) locally innervated by distinct IN subtypes (an IN-PN circuit). In the neocortex, PNs are grouped by areas, layers, and remote projection targets, which represent their functional attributes. It has been shown that distinct classes of PNs display unique homotypic- and heterotypic-connections and convey different neuronal signals. However, little is known about cellular and axonal organization of distinct IN subtypes sending inputs to defined PNs. To address this question, we have developed a novel genetic strategy combining rabies virus (RV)-mediated retrograde monosynaptic labeling and intersectional approaches. The major objective of our proposal is to provide wiring and developmental principles of INs sending inputs to defined PN subtypes at a cell type-specific resolution. Previous studies showed that layer 5 (L5) PNs receive a larger number of inhibitory inputs from parvalbumin (PV)-expressing INs than L 2/3 PNs and this connection feature is controlled by PN identity. Thus, we hypothesize that distinct PN types defined by areas, layers, and long-range projection targets have different organization of input INs, which is shaped at least in part by PN identity. To test this hypothesis, we will dissect the following subjects using an intersectional retrograde monosynaptic tracing, genetic manipulation of PN identity, and mouse genetics. In Aim 1, we will elucidate organization of PV-, somatostatin (SOM)-, or vasoactive intestinal polypeptide (VIP)-expressing INs sending inputs to distinct PN types defined by cortical areas, laminar positions, and remote projection targets. In Aim 2, we will examine developmental processes of IN-PN circuits containing specific PN types innervated by PV-, SOM-, or VIP-INs. In Aim 3, we will generate ectopic PNs by genetic manipulations of transcription factors that control PN identities and examine organization of input IN subtypes. Through these experiments, we will gain wiring and developmental principles of IN-PN circuits in a cell type-specific manner, which will pioneer novel approaches for diagnosis and treatment of brain disorders.

项目总结/摘要 GABA能中间神经元（INs）形成的抑制回路参与皮层神经元的加工和编码， 信息通过塑造神经活动的空间和时间结构。与这一关键角色相一致 由于IN在正常脑功能中的重要性，IN功能障碍与多种脑功能障碍有关， 比如精神分裂症自闭症和癫痫尽管它们很重要，但抑制性局部神经元的详细接线图 由于IN类型的巨大多样性，电路在很大程度上仍然未知。此外，人们也不太了解 原理支配抑制性微电路的组装。填补这些知识空白将为我们提供 皮层抑制回路的布线和发展原则，这反过来又极大地促进了我们的大脑。 大脑皮层回路是如何工作的一个基本的皮层回路模块包含一个兴奋性的 主神经元（PN）局部受不同IN亚型（IN-PN回路）支配。在新皮层中， 按区域、层和远程投影目标分组，这些目标表示其功能属性。它有 已经表明，不同类别的PN显示独特的同型和异型连接，并传达 不同的神经信号然而，对不同IN的细胞和轴突组织知之甚少， 子类型将输入发送到定义的PN。为了解决这个问题，我们开发了一种新的基因 结合狂犬病病毒（RV）介导逆行单突触标记和交叉策略 接近。我们建议的主要目标是提供智能网的布线和发展原则 以小区类型特定的分辨率向定义的PN子类型发送输入。先前的研究表明， 5（L5）PN比L2/3 PN接受更多来自表达小白蛋白（PV）的IN的抑制性输入 PN，并且该连接特征由PN身份控制。因此，我们假设不同的PN类型 由区域、图层和远程投影目标定义的输入IN具有不同的组织， 至少部分地由PN标识来成形。为了验证这一假设，我们将使用一个 交叉逆行单突触追踪、PN身份的遗传操作和小鼠遗传学。在 目的1、阐明PV-、生长抑素（SOM）-或血管活性肠肽的组织结构 （VIP）-表达IN向由皮质区、椎板位置和 远程投射目标在目标2中，我们将研究IN-PN电路的发展过程， 由PV-、SOM-或VIP-IN支配的特定PN类型。在目的3中，我们将通过遗传学方法产生异位PN。 操纵控制PN身份和检查输入IN组织的转录因子 亚型通过这些实验，我们将获得IN-PN电路的布线和开发原则， 细胞类型特异性的方式，这将开创新的方法，诊断和治疗脑 紊乱

项目成果

In Vivo Single-Cell Genotyping of Mouse Cortical Neurons Transfected with CRISPR/Cas9.

转染 CRISPR/Cas9 的小鼠皮质神经元的体内单细胞基因分型。

• DOI: 10.1016/j.celrep.2019.06.038
• 发表时间: 2019
• 期刊: Cell reports
• 影响因子: 8.8
• 作者: Steinecke,André;Kurabayashi,Nobuhiro;Hayano,Yasufumi;Ishino,Yugo;Taniguchi,Hiroki
• 通讯作者: Taniguchi,Hiroki