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-实验程序，以定量癫痫发作活动与神经元的分辨率在完整的幼斑马鱼中枢神经 系统 我们的研究是可能的光片显微镜的最新进展，以及在大型神经成像数据集分析的理论和算法的进步。 光片显微镜具有良好的空间和时间分辨率，是能够几乎完全的体积覆盖的幼虫斑马鱼中枢神经系统。 这一点，沿着先进的统计和计算技术，使我们能够以前所未有的准确度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围绕兴奋区域，保护大脑免受癫痫发作时产生的失控兴奋性（发作）活动的影响。 然而，反复的发作活动波可以打破周围的抑制，使癫痫发作传播。 通过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（通常称为“抑制性抑制”）相互作用，这是大脑对癫痫发作的反应。 一个精确的理解如何连贯的兴奋相互作用，抑制interneuron人口将提供一个强大的控制范式的空间和时间干预癫痫发作的形成和传播。 此外，虽然已经使用来自解剖数据的详细突触连接重建进行了计算机模拟，但在同一生物体中导出然后验证的模拟将对癫痫发作的研究以及更普遍的神经科学做出变革性贡献。 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到一个统一的整体中，研究了一个大型，完整的神经元网络。 虽然由于技术原因，这种方法仅限于斑马鱼幼虫，一种小而透明的生物，但它可以从根本上提高我们对脊椎动物中中尺度神经元系统保护机制如何失败的理解。 我们提出的研究将提供信息，可以指导未来的癫痫发作干预，如神经元移植，电刺激，手术组织切除或药物靶向神经元群体和突触，最有效地防止癫痫发作的形成和传播。 参与我们计划的研究生和博士后的教育和培训将与研究的各个方面相结合。 本科生将参与研究和指导。 这项研究是多机构的，建立在工程，发育神经科学，癫痫和数学现有的跨学科合作基础上。

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.