Development of cortical circuits using multiscale optical imaging in awake mice

使用多尺度光学成像在清醒小鼠中开发皮层回路

• 批准号: 9611652
• 负责人: Ali Saddam Hamodi Alshuwaykh
• 金额: $ 6.14万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2021-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9611652
• 关键词: Adult; Amblyopia; Architecture; Autistic Disorder; Behavior; Birth; Brain; Calcium; Cells; Chronic; Complex; Coupling; Custom; Development; Electrophysiology (science); Epilepsy; Equilibrium; Esthesia; Exhibits; Eye; Frequencies; Functional disorder; Ganglia; Gap Junctions; Generations; Glutamates; Growth; Head; Image; Interneurons; Investigation; Maps; Measures; Medial; Mediating; Microscope; Microscopic; Molds; Molecular; Morphology; Mus; Neuraxis; Neurodevelopmental Disorder; Neurons; Pathologic; Pattern; Perinatal; Photic Stimulation; Photons; Play; Population; Population Heterogeneity; Process; Property; Reporter; Reporting; Resolution; Retina; Retinal; Role; Sensory; Shapes; Signal Transduction; Somatostatin; Sorting - Cell Movement; Speed; Strabismus; Testing; Transgenic Mice; Vasoactive Intestinal Peptide; Viral; Viral Vector; Vision; Visual; Visual Cortex; Visual system structure; Work; awake; calcium indicator; cell type; cholinergic; excitatory neuron; imaging approach; inhibitory neuron; microscopic imaging; neural circuit; neural patterning; neurodevelopment; neuronal circuitry; neurotransmission; novel; optical imaging; orientation selectivity; promoter; recombinase-mediated cassette exchange; recruit; relating to nervous system; retinotopic; spatiotemporal; starburst amacrine cell; tool; two-photon; visual map; visual processing; visual stimulus

PROJECT SUMMARY Development of neural circuits depends on a combination of molecular and activity-dependent processes. The activity can be spontaneous—such as retinal waves that course across the mammalian perinatal retina—or sensory-evoked. Spontaneous waves of retinal activity are necessary for circuit refinement in the brain. Although much is known about the retinal circuitry responsible for retinal wave generation, it is unclear how retinal waves shape the development of functional cortical architecture and what role they play in the emergence of functional cortical properties that are already established at eye opening, such as retinotopy and orientation selectivity. It is also completely unknown which subtypes of neurons participate in waves in visual cortex and when they are recruited during development. Here we will determine the roles and dynamics of excitatory and inhibitory neuron activity in visual cortex during the period of retinal waves using simultaneous cellular-resolution two- photon calcium imaging of genetically defined neurons within a local circuit and simultaneous wide field single- photon imaging of neuronal activity across cortex to assess the function of retinal waves in cortical circuit formation in awake, head-fixed mice. This novel multiscale, dual-imaging approach will bridge the gap between microscopic and macroscopic imaging in order to better understand the relationship between local cellular activity and mesoscale, cortex-wide activity. To identify interneurons and measure their participation in retinal wave activity in cortex, we will express the fluorescent reporter tdTomato in inhibitory interneurons using the cre/lox system in transgenic mouse lines expressing the green calcium indicator GCaMP6s under the pan- neuronal snap25 promotor. We will also assess the ontogeny of the spatiotemporal activity pattern of Nkx2.1-, somatostatin- (SOM), and vasointestinal peptide-expressing (VIP) interneurons and excitatory/inhibitory coupling on the mesoscale level during stage II and stage III retinal wave activity in visual cortex using viral expression of jRCaMP1b in interneurons and GCaMP6s in excitatory neurons. We hypothesize that cortical interneurons shape the flow of retinal wave activity in visual cortex and contribute to the emergence of retinotopy and orientation selectivity. We will test this by chronically silencing these cells using GiDREADD, then assessing spatiotemporal dynamics of retinal wave generated activity in visual cortex, in addition to mapping retinotopy and measuring orientation tuning using visual stimulation at eye opening. Furthermore, we will determine the specific roles of Nkx2.1, SOM, VIP interneurons during stage II and stage III retinal wave activity in visual cortex. This work will develop and apply essential tools for detailed investigation of the impact of specific neuronal populations on the functional organization of brain circuits, and will describe how the developing cortex propagates spontaneous activity, and how the functional connectivity between different subtypes of cortical neurons is sculpted by early spontaneous activity. This is critical to understanding normal brain development and dysfunction associated with neurodevelopmental disorders.

项目摘要 神经回路的发展取决于分子和活动依赖性过程的组合。的 活动可以是自发的--例如穿过哺乳动物围产期视网膜的视网膜波--或者 感官诱发视网膜活动的自发波对于大脑回路的完善是必要的。虽然 关于负责视网膜波产生的视网膜回路已经知道很多，但不清楚视网膜波是如何产生的。 塑造功能性皮层结构的发展，以及它们在功能性皮层结构的出现中扮演什么角色。 在睁眼时已经建立的皮质特性，如视网膜反射和方向选择性。它 也完全不知道哪些神经元亚型参与视觉皮层的波，以及它们何时参与 在开发过程中招募。在这里，我们将确定兴奋性和抑制性的作用和动力学 用同步细胞分辨双通道技术研究视网膜波期间视皮层神经元的活动。 局部回路内遗传定义的神经元的光子钙成像和同时宽场单光子钙成像， 应用光子成像技术检测皮层神经元活动以评价视网膜波在皮层回路中的功能 在清醒的头部固定的小鼠中形成。这种新颖的多尺度双成像方法将弥合 为了更好地理解局部细胞之间的关系， 活动和中尺度的，皮层范围的活动。为了识别中间神经元并测量它们在视网膜神经元中的参与， 波的活动，我们将表达荧光报告tdTomato在抑制性中间神经元使用 cre/lox系统在表达绿色钙指示剂GCaMP 6 s的转基因小鼠系中的pan- 神经元snap 25启动子。我们还将评估Nkx2.1-的时空活动模式的个体发生， 生长抑素（SOM）和血管肠肽表达（VIP）的中间神经元和兴奋/抑制 使用病毒在视皮层中的II期和III期视网膜波活动期间在中尺度水平上的耦合 jRCaMP 1b在中间神经元和GCaMP 6s在兴奋性神经元的表达。我们假设大脑皮层 中间神经元塑造视皮层中视网膜波活动的流动，并有助于视网膜病变的出现 和取向选择性。我们将通过使用GiDREADD长期沉默这些细胞来测试，然后评估 视网膜波的时空动力学在视皮层产生活动，除了映射视网膜病变 以及在睁眼时使用视觉刺激来测量定向调谐。此外，我们将确定 Nkx2.1、SOM、VIP中间神经元在视皮层II期和III期视网膜波活动中的特定作用。 这项工作将开发和应用必要的工具，详细调查特定神经元的影响， 人口对大脑回路的功能组织，并将描述如何发展皮层 传播自发活动，以及不同皮质亚型之间的功能连接如何 神经元是由早期自发活动塑造的。这对理解正常的大脑发育至关重要 以及与神经发育障碍相关的功能障碍。