CRCNS: Quantitation of Network Dysfunction in Epilepsy-Understanding the Inhibitory Restraint

描述(由申请人提供)：我们提出了一个理论-实验方案来量化完整的斑马鱼幼体中枢神经中神经元分辨率的癫痫发作活动。 系统。我们的研究是由于光片显微镜的最新进展，以及在分析大型神经成像数据集方面的理论和算法进步而成为可能的。光片显微镜具有良好的空间和时间分辨率，能够几乎完全覆盖斑马鱼幼体中枢神经系统的体积。这一点，加上先进的统计和计算技术，使我们能够以前所未有的准确性量化斑马鱼大脑中的神经动力学。由于神经回路的结构，抑制性神经元群体通常环绕在兴奋区域，保护大脑免受癫痫形成时产生的失控兴奋(发作)活动的影响。然而，发作活动的重复波可以打破周围的抑制，允许癫痫发作的传播。利用高分辨率显微镜和最先进的计算分析和模拟方法，我们将研究癫痫发作期间产生的连贯发作活动如何与周围抑制(通常被称为“抑制抑制”)相互作用，这是大脑对癫痫的反应。准确理解相干兴奋如何与神经元间抑制相互作用和抑制，将为癫痫形成和传播的空间和时间干预提供强有力的控制范例。此外，尽管已经使用从解剖数据重建详细的突触连接性进行了计算机模拟，但从同一生物体中推导并验证的模拟将对癫痫研究乃至更广泛的神经科学做出革命性的贡献。这项研究首次将神经科学研究的实验、理论和验证方面结合成一个统一的整体，研究一个大型、完整的神经元网络。尽管由于技术原因，这种方法仅限于斑马鱼幼体，一种小型、透明的生物，但它可以从根本上提高我们对中尺度神经系统的保护机制如何在脊椎动物中失效的理解。我们拟议的研究将提供信息，以指导未来的癫痫干预措施，如神经元移植、电刺激、外科组织切除或最有效地防止癫痫形成和传播的神经元和突触的药物靶向。我们项目涉及的研究生和博士后的教育和培训将融入研究的方方面面。本科生将参与研究并进行指导。这项调查是多机构的，建立在工程学、发育神经科学、癫痫和数学领域现有的跨学科合作的基础上。

DESCRIPTION (provided by applicant): We propose a theoretical-experimental program to quantitate seizure activity with neuronal resolution in the intact larval zebrafish central nervous system. Our study is made possible by recent advances in light-sheet microscopy, and theoretical and algorithmic advances in the analysis of large neural imaging datasets. Light-sheet microscopy has both excellent spatial and temporal resolution and is capable of virtually complete volumetric coverage of the larval zebrafish central nervous system. This, along with advanced statistical and computational techniques, allows us to quantify neural dynamics in the zebrafish brain with unprecedented accuracy. Because of the structure of neural circuits, inhibitory neuronal populations typically surround excited regions, protecting the brain from runaway excitatory (ictal) activity that is generated when a seizure forms. However, repeated waves of ictal activity can break down the surround inhibition, allowing a seizure to propagate. With high-resolution microscopy and state-of-the-art computational analysis and simulation methods, we will study how coherent ictal activity generated during seizures interacts with the surround inhibition (often called the 'inhibitory restraint') that is the brain's response to the seizure. A precise understanding of how coherent excitations interact with and depress inhibition in interneuron populations would provide a powerful control paradigm for spatial and temporal intervention in seizure formation and propagation. Furthermore, although computer simulations have been performed using detailed synaptic connectivity reconstructions from anatomical data, simulations derived, then validated in the same organism would be a transformative contribution to the study of seizures and more generally to neuroscience. This study combines the experimental, theoretical, and validation aspects of a neuroscience investigation into a unified whole in the study of a large, intact neuronal network for the first time. Although, for technical reasons, this approach is limited to the larval zebrafish, a small, transparent organism, it could radically improve our understanding of how protective mechanisms in meso-scale neuronal systems can fail in vertebrates. Our proposed study will provide information that could guide future seizure interventions such as neuron transplantation, electrical stimulation, surgical tissue removal or drug targeting of neuronal populations and synapses that most effectively prevent seizure formation and propagation. The education and training of the graduate students and postdocs involved in our program will be integrated with every aspect of the research. Undergraduate students will be involved in the research and mentored. The investigation is multi-institutional and builds on existing interdisciplinary collaborations in engineering, developmental neuroscience, epilepsy and mathematics.