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性依赖性过程的结合。这 活动可以是发起的，例如跨哺乳动物围产期视网膜的残留波，或 感官诱发的。自发性视网膜活性波是大脑中电路的细化所必需的。虽然 对于负责视网膜波的负责的视网膜电路知之甚少，目前尚不清楚视网膜波如何 塑造功能性皮质结构的发展及其在功能的出现中的作用 已经在目光开口中建立的皮质特性，例如视网膜和方向选择性。它 也完全未知，哪些神经元的亚型参与视觉皮层中的波 在开发过程中招募。在这里，我们将确定兴奋和抑制的角色和动力学 在残留波的期间，使用简单的细胞分辨率二 - 局部电路中普遍定义的神经元的光子钙成像和简单的宽场单个 跨皮质的神经元活性的光子成像，以评估视网膜波的功能 醒着，头部固定的小鼠形成。这种新颖的多尺度，双成像方法将弥合 显微镜和宏观成像，以更好地了解局部细胞之间的关系 活动和中尺度，皮质范围的活性。识别中间神经元并衡量他们参与残差 在皮质中的波活性，我们将使用抑制性中间神经元中的荧光记者TDTOMATO表示 转基因小鼠系中的CRE/LOX系统，表达绿色钙指示剂GCAMP6s 神经元SNAP25启动子。我们还将评估NKX2.1-的时空活性模式的个体发育。 生长抑素 - （SOM）和过量表达肽（VIP）中间神经元和兴奋性/抑制性 在II期和III期在视觉皮层中使用病毒的耦合在II期和III期视网膜波活动期间的耦合 JRCAMP1b在兴奋性神经元中的神经元和GCAMP6中的表达。我们假设皮质 中间神经元塑造视觉皮层中视网膜波活性的流动，并导致视网膜的出现 和方向选择性。我们将通过使用GidreadD长期沉默这些单元进行测试，然后评估 除了映射视网膜外，视网膜波生成活性的时空动力学 并使用视觉刺激在眼开刺激下测量方向调整。此外，我们将确定 II期和III期视网膜波活性在视觉皮层中的NKX2.1，SOM，VIP中间神经元的特定作用。 这项工作将开发并应用基本工具，以详细研究特定神经元的影响 大脑电路功能组织的人群，并将描述发育中的皮层 传播赞助商活动，以及皮质不同亚型之间的功能连通性 神经元是由早期赞助者活动雕刻的。这对于理解正常的大脑发育至关重要 与神经发育障碍相关的功能障碍。