Massively scalable 3D electrophysiology and two-photon imaging in freely-moving animals

自由移动动物的大规模可扩展 3D 电生理学和双光子成像

• 批准号: 10687565
• 负责人: Krishna jayant
• 金额: $ 130.42万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10687565
• 关键词: 3-Dimensional; Animals; Behavior; Behavioral; Biological Assay; Brain; Calcium; Custom; Electrodes; Electronics; Electrophysiology (science); Environment; Head; Height; Image; Learning; Maps; Microscope; Microscopic; Motion; Movement; Mus; Nature; Neurosciences; Preparation; Resolution; Sensory; Shapes; Signal Transduction; Sleep; Stimulus; Structure; Synapses; Tactile; Touch sensation; Travel; Vibrissae; awake; brain tissue; brain volume; density; design; experience; experimental study; flexibility; innovation; memory consolidation; memory process; millisecond; nanoelectrode array; nanoelectrodes; neural circuit; sensory discrimination; sensory system; technology platform; two-photon

SUMMARY Revealing how neural circuits encode and enable behavioral experiences is a fundamental problem in neuroscience. Decoding and dissecting the mechanisms of signal flow across such circuits necessitates the ability to record millisecond electrical dynamics and simultaneously map the spatial organization of cellular and sub-cellular circuit motifs, in awake behaving animals. During natural behavior, animals actively acquire sensory information as they move through the environment and use this information to guide ongoing actions. In this context, unconstrained movement-related signals could allow sensory systems to efficiently predict self- generated motion and extract additional information about the environment, thereby forming a stable internal representation of the external world. However, a majority of recordings are performed in head-fixed animals which imposes severe restrictions on how movement related signals shape ongoing sensory and memory processing in the brain. Performing high-density electrophysiology and concomitant two-photon calcium imaging is -at present- not feasible due to technical limitations and are therefore performed separately in both head fixed and freely moving preparations. There is a great need for technology platforms that can combine high-resolution electrical recordings across entire volumes of brain tissue and two-photon calcium imaging in freely behaving animals. In this proposal we introduce a new paradigm for high-density electrophysiology across 3D volume with capabilities to simultaneously perform two-photon calcium imaging. Our innovation termed NET-2P, comprises of 3D Nanoelectrodes of variable height integrated onto the baseplate of a head-mounted mini two-photon microscope. The Nanoelectrode array is integrated with custom-designed CMOS electronics and will in total weigh 3.5 grams. The transparent and flexible nature of the nanoelectrode array allows for easy two-photon access whilst facilitating rapid electrical mapping across large cortical sections. We propose to use the head- mounted setup to 1) assay cortical travelling waves under tactile processing whilst mapping the underlying cellular scale ensemble map via two-p imaging and 2) unravel how cortical ensembles and travelling waves that emerge after a learning task enhance memory consolidation during sleep. In preliminary experiments performed in head-fixed awake animals under passive whisker touch, we used planar transparent electrode array recordings and conventional two-photon calcium imaging, and discovered microscopic travelling waves upon whisker touch, a late reverberatory wave 100ms post touch, and sparse yet stable cellular ensemble structure that supports wave propagation. We hypothesize that spontaneous travelling waves, including late reverberatory components that emerge hundreds of milliseconds post stimulus, carry movement related, head-direction, and volitional control signals, which will enhance travelling wave dynamics in freely-moving mice. By combining electrophysiology and imaging-based ensemble mapping during natural sleep we will assay how spiking and synaptic changes across cortical layers strengthen and enable robust functional cellular activity landscapes.

摘要 揭示神经回路如何编码和启动行为体验是 神经科学。解码和剖析通过这类电路的信号流的机制需要 能够记录毫秒级的电动力学，并同时绘制细胞和 在清醒的行为动物中，亚细胞环路图案。在自然行为中，动物主动地获得感官 他们在环境中移动时的信息，并使用这些信息来指导持续的行动。在这 背景下，不受限制的运动相关信号可以使感觉系统有效地预测自我 生成运动并提取有关环境的附加信息，从而形成稳定的内部 外部世界的表现。然而，大多数录音是在头部固定的动物身上进行的 这对运动相关信号如何塑造正在进行的感觉和记忆施加了严格的限制 在大脑中进行处理。进行高密度电生理和伴随的双光子钙成像 由于技术限制，目前是不可行的，因此在两个固定的头部分别进行手术 和自由移动的制剂。非常需要结合高分辨率的技术平台 全脑组织的电记录和自由活动状态下的双光子钙成像 动物。在这项建议中，我们引入了一种新的跨3D体积的高密度电生理学范例 能够同时执行双光子钙成像。我们的创新名为Net-2P，包括 将高度可变的3D纳米电极集成到头戴式微型双光子的基板上 显微镜。纳米电极阵列与定制设计的cmos电子设备集成在一起，将总共 体重3.5克。纳米电极阵列的透明和灵活的性质允许容易的双光子 访问，同时促进在大脑皮层切片上的快速电子标测。我们建议使用头部- 安装装置以1)在触觉处理下分析皮质行波，同时标测潜在的 通过双P成像的细胞尺度集合图以及2)揭示大脑皮层集合和行波如何 完成一项学习任务后出现，增强睡眠期间的记忆巩固。在进行的初步实验中 在头部固定的清醒动物中，我们使用了平面透明电极阵列 记录和常规的双光子钙成像，并在 胡须触摸，触摸后100毫秒的延迟反射波，稀疏但稳定的细胞整体结构 这支持了波的传播。我们假设包括延迟反射在内的自发行波 刺激后数百毫秒出现的部件，携带与运动相关的、头部方向和 意志力控制信号，这将增强自由活动小鼠的行波动力学。通过组合 在自然睡眠期间，我们将分析电生理和基于成像的整体标测如何产生尖峰电位和 跨大脑皮层的突触变化加强并使强大的功能细胞活动图景。