Multiscale Imaging of Spontaneous Activity in Cortex: Mechanisms, Development and Function

 描述(申请人提供)：本RFA的目的是促进神经回路和系统检查的实验、分析和理论能力的整合。这项提议对RFA反应迅速，因为它连接了几个不同的神经科学实验室，以开发提供同时多状态成像的新技术，并将这些技术应用于检查哺乳动物皮质中的神经元动力学如何随着大脑状态和发育的变化而变化。配对成像模式将弥合从单个神经元的成像活动到整个大脑电路水平分析的差距。不同的尺度将与一个全面的模型联系在一起，这样每一级实验都可以相互启发。我们将开发这项技术，以允许局部电路的单细胞(双光子)钙离子成像和整个大脑皮层的中观(单光子)钙离子成像。在同一动物的单独配对研究中，我们将利用全脑功能磁共振成像，在大脑皮层同时进行中观钙离子成像。全皮质介观钙成像是由其中一家PI(CRAIR)开发的一项创新技术，该技术使用在所有神经元或特定神经元群体中表达遗传编码的钙指示器(GCaMP6)的小鼠，允许在完整、未麻醉的发育中的小鼠大脑中进行局部神经元和经颅种群水平的跨皮质介观尺度成像。这项钙离子成像技术将使我们能够直接将单细胞成像与大脑皮层和整个大脑的总回路水平的活动联系起来。为了了解宏观回路变化的神经来源，以及通过发育和作为行为脑状态的函数影响这一组织的因素，提出了一个将这些不同模式联系起来的综合模型。这项工作在有待进一步开发的新型钙离子成像策略以及在配对尺度成像的设计上具有创新性，以建立单个神经元活动与电路水平组织之间的联系。这项工作意义重大，因为它将为详细研究特定神经元细胞群在健康发育和疾病模型中对脑回路功能组织的影响提供一套工具。同样重要的是，对神经元活动的来源和流动的新见解将提高我们对指导大脑发育中的自组织及其通过大脑状态进行动态调节的原理的理解。

 DESCRIPTION (provided by applicant): The purpose of this RFA is to promote the integration of experimental, analytic and theoretical capabilities for the examination of neural circuits and systems. This proposal is highly responsive to the RFA in that it links several different neuroscience labs to develop new technologies that provide for simultaneous multistate imaging and applies these technologies to the examination of how neuronal dynamics in mammalian cortex varies as a function of brain state and development. Paired imaging modalities will bridge the gap from imaging activity in individual neurons to whole brain circuit level analyses. The different scales will be linked with a comprehensive model such that each level of experimentation can inform the other. We will develop the technology to allow simultaneous single cell (two-photon) Ca2+ imaging of a local circuit and cortex-wide mesoscopic (single-photon) Ca2+ imaging across the entire neocortex. In separate paired studies in the same animals we will develop simultaneous mesoscopic Ca2+ imaging across the cortex with whole-brain functional MRI. Whole cortex mesoscopic Ca2+ imaging represents an innovative technology developed by one of the PIs (Crair) that uses mice expressing a genetically encoded Ca2+ indicator (GCaMP6) in all neurons or in select populations of neurons to allow both local neuronal and transcranial population level mesoscopic scale imaging across the cortex in the intact, unanesthetized developing mouse brain. This Ca2+ imaging technique will allow us to directly link single cell imaging to gross circuit level activity across the cortex and whole brain An integrative model is proposed to link these different modalities in order to understand the neural source of macroscopic circuit changes and the factors that influence this organization through development and as a function of behavioral brain state. This work is innovative in the novel Ca2+ imaging strategies to be further developed, and in the design of paired scale imaging to establish links between single neuron activity and circuit level organization. The work is significant in that it will provide a set of tools for detailed investigations of the impact of specific neuronal cell populations on brain circuit functional organization in healthy development and disease models. It is also significant in that new insights into the source and flow of neuronal activity will be obtained that will improve our understanding of the principles guiding self-organization in the developing brain and its dynamic modulation by brain state.