Real-time, all-optical interrogation of neural microcircuitry in the pretectum

摘要 理解神经回路如何引起行为的主要障碍之一是， 实验性的准备工作使得研究这些跨不同脑区的回路变得困难。最近 显微镜和钙传感器的进步使得同时记录多达 数以千计的单个神经元，光学方法使得一次刺激数百个神经元成为可能。 但是目前的方法只刺激预先确定的神经元的子集， 用于解剖大规模神经回路。在这里，我们建议开发一种新的综合实验- 一个测试斑马鱼视动反应神经回路假设的计算平台， 代表性的感觉运动行为。这个平台将使我们能够描述 功能上定义的神经元组作为实时收集的数据。通过使用事先指导的 自适应地选择多达数百种的无扫描3D全息光刺激图案的算法， 神经元响应先前观察到的数据，我们将能够以指数方式增加数据 效率，同时推断视觉响应之间的多种功能连接 神经元在斑马鱼pretectum和他们的下游目标。一旦建立，这种方法将 允许我们进行适应性实验，根据功能选择性地干扰神经功能， 加速模型生成和假设检验的过程。此外，这些工具将 适用于其他类型的钙成像数据，对系统神经科学具有广泛的影响。

Abstract One of the major barriers to understanding how neural circuits give rise to behavior is that typical experimental preparations make it difficult to study these circuits across different brain areas. Recent advances in microscopy and calcium sensors have made it possible to simultaneously record up to thousands of individual neurons, and optical methods have made it possible to stimulate hundreds at a time, but current approaches, which stimulate only subsets of predetermined neurons, are not adequate for dissecting large-scale neural circuits. Here, we propose to develop a novel integrated experimental- computational platform to test neural circuit hypotheses of the zebrafish optomotor response, a representative sensorimotor behavior. This platform will allow us to characterize the relationships among functionally defined groups of neurons as the data are collected in real-time. By using prior-guided algorithms that adaptively choose scanless 3D holographic photostimulation patterns of up to hundreds of neurons in response to previously observed data, we will be able to exponentially increase data efficiency, simultaneously inferring multiple classes of functional connections between visually responsive neurons in the zebrafish pretectum and their downstream targets. Once established, this approach will allow us to perform adaptive experiments that selectively perturb neural function based on function, accelerating the process of model generation and hypothesis testing. Moreover, these tools will be applicable to other types of calcium imaging data, with broad implications for systems neuroscience.