Optogenetic Tools for in vivo Analysis of Cortical Circuit Plasticity

用于皮层回路可塑性体内分析的光遗传学工具

基本信息

批准号：
7914333
负责人：
PETER SAGGAU
金额：
$ 58.88万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-08-15 至 2012-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7914333
关键词：
3-Dimensional Adult Behavior Biological Neural Networks Brain Cells Cerebral cortex Data Development Dimensions Electroporation Functional Imaging Glutamates Goals Image In Vitro Individual Injury Ion Channel Knowledge Label Lasers Learning Maps Measures Methods Microscope Microscopy Molecular Monitor Mus Neuronal Plasticity Neurons Optics Pattern Plastics Population Process Property Proteins Rabies virus Radial Ramp Research Research Infrastructure Research Personnel Resolution Sampling Scanning Sensory Shadowing (Histology)Shapes Site Slice Source Speed Substance of Abuse Surface Synapses System Technology Time Tracer Translating Viral Visual Cortex experience imaging modality in vivo interest light microscopy method development multi-photon neural circuit novel photolysis plasmid DNA presynaptic relating to nervous system response skills tool two-photon

项目摘要

Neuroplasticity is central to fundamental processes in the brain, including learning, and long-lasting changes in neural circuits that result from employing substances of abuse. In contrast to significant advances at the cellular/molecular level, our understanding of the functional organization and reorganization of brain circuits remains limited. Increasing in vivo evidence suggests that single cells remain plastic into and during adulthood. However, our knowledge of the functional properties of single cells is primarily descriptive. This limits our understanding of the underlying mechanisms involved in assembling and modifying neuronal tuning functions during plasticity. In order to understand how the properties of cells change during plasticity, it is imperative to record from a population of interconnected neurons in vivo. More than half of all synaptic contacts in the cortex arise from neurons within a -200 j.lm radius from the target cell. Importantly, connections between cells in the cerebral cortex are predominantly along cortical depth. Therefore, we need to monitor simultaneously the activity of all adjacent neurons in a cortical volume, and thus record in three dimensions. To date, no experimental tool exists that would allows us to do this. This project seeks to establish novel in vivo methods that will allow us to analyze neural circuits in three dimensions. For this purpose, we will advance two technologies to record from and manipulate circuits in the mammalian cortex: (1) Ultra-fast three-dimensional two-photon imaging, and (2) Optical manipulation of neural activity with single-cell and single-spike resolution using optogenetic tools. The proposed developments have become possible because of recent technical advances in ultra-fast multi-photon microscopy and light-activated ion channels. In short: Inertia-free nearinfrared laser scanning technology allows for in vivo fast structural and functional imaging as well as for high resolution optical stimulation. The proposed project will combine these key technologies to generate the infrastructure and the experimental skills to study the function and plasticity of cortical microcircuits and thus will help researchers to understand mechanisms of neuroplasticity in the intact brain.

神经可塑性是大脑基本过程的核心，包括学习以及因使用滥用物质而导致的神经回路的长期变化。与细胞/分子水平上的重大进展相比，我们对大脑回路的功能组织和重组的理解仍然有限。越来越多的体内证据表明，单细胞在成年期和成年期仍保持可塑性。然而，我们对单细胞功能特性的了解主要是描述性的。这限制了我们对可塑性期间组装和修改神经元调节功能所涉及的潜在机制的理解。为了了解细胞的特性在可塑性过程中如何变化，必须记录体内互连神经元群的情况。皮质中超过一半的突触接触来自目标细胞 -200 j.lm 半径范围内的神经元。重要的是，大脑皮层细胞之间的连接主要沿着皮层深度。因此，我们需要同时监测皮质体积中所有相邻神经元的活动，从而进行三维记录。迄今为止，还没有实验工具可以让我们做到这一点。该项目旨在建立新颖的体内方法，使我们能够在三个维度上分析神经回路。为此，我们将推进两项技术来记录和操纵哺乳动物皮层中的电路：（1）超快三维双光子成像，以及（2）用单细胞和单细胞对神经活动进行光学操纵。使用光遗传学工具进行尖峰分辨率。由于超快多光子显微镜和光激活离子通道的最新技术进步，所提出的发展已成为可能。简而言之：无惯性近红外激光扫描技术可实现体内快速结构和功能成像以及高分辨率光学刺激。拟议的项目将结合这些关键技术来生成研究皮质微电路的功能和可塑性的基础设施和实验技能，从而帮助研究人员了解完整大脑中的神经可塑性机制。