Development of an integrated array for simultaneous optogenetic stimulation and electrical recording to study cortical circuit function in the non-human primate brain

开发用于同步光遗传学刺激和电记录的集成阵列，以研究非人类灵长类动物大脑中的皮质电路功能

• 批准号: 9358355
• 负责人: Alessandra Angelucci
• 金额: $ 74.97万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-30 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9358355
• 关键词: Ablation; Acute; Address; Animals; Architecture; Area; Autistic Disorder; Back; Behavior; Behavioral; Biological Models; Brain; Brain Diseases; Caliber; Cerebral cortex; Chronic; Coupling; Development; Devices; Electrodes; Electrophysiology (science); Engineering; Epilepsy; Foundations; Functional disorder; Future; Generations; Glass; Goals; Histologic; Human; Hybridization Array; Industrialization; Investigation; Lasers; Lead; Learning; Light; Lighting; Link; Macaca; Measurable; Medical; Mental disorders; Methods; Morphologic artifacts; Mus; Neuroanatomy; Neurologic; Neurons; Neurophysiology - biologic function; Neurosciences; Opsin; Optics; Pattern; Perception; Physiological; Physiology; Plasticizers; Positioning Attribute; Primates; Proteins; Research; Resources; Schizophrenia; Side; Study models; Surface; Technology; Testing; Therapeutic Intervention; Time; Tissues; Universities; Utah; Visible Radiation; Visual Cortex; Work; attenuation; awake; base; biomaterial compatibility; cell type; commercialization; design; experimental study; in vivo; microbial; microsystems; millisecond; nervous system disorder; neural circuit; neurophysiology; neuroregulation; new technology; nonhuman primate; optical fiber; optogenetics; photoactivation; photonics; prevent; prosthesis control; prototype; relating to nervous system; spatiotemporal; tool

Understanding the function of neural circuits in the cerebral cortex of the non-human primate (NHP), the model system closest to human, is crucial to understanding normal cortical function and the circuit-level basis of human brain disorders. Optogenetics has emerged as a powerful tool for studying neural circuit function, by using light to perturb the activity of specific cell types genetically modified to express light-activated microbial opsins, and assessing the consequences of this perturbation on network activity and behavior. While successful in mice, it has been challenging to apply optogenetics to NHPs, largely due to the lack of multifunction integrated probes for precision light delivery and electrophysiology across mm-to-cm volumes through the depth of the NHP cortex. Large volume manipulations are essential in the large NHP brain in order to observe measurable electrophysiological or behavioral effects. An interdisciplinary team of PIs proposes to develop and test in vivo integrated penetrating arrays that allow for large-volume, spatiotemporally patterned optogenetic modulation and electrical recording of neural circuits in the NHP brain. This project requires the coordinated effort of 4 teams, including experts in photonic devices and µLED development for optogenetics, materials and packaging for biocompatible devices, primate neurophysiology, and pioneers in electrode array design and commercialization. In Aim 1 we develop the technology, and in Aim 2 we test it in vivo in the NHP visual cortex. We will initially develop a 4x4 mm penetrating 10x10 optrode array in a format analogous to the well- established Utah Electrical Array (UEA), with each probe serving as a waveguide allowing visible light to reach tissue depths >1.5mm. Following initial optimization of the probe's shank diameter and tip angle to minimize tissue damage, we will perform proof-of-concept in vivo NHP optogenetic experiments in deep cortical tissue, using broad-area illumination of the entire array. In a second stage, we will develop light coupling via µLEDs, which will be integrated into a single platform and tested in vivo, consisting of a µLED located over each optical probe. Completion of stage 2 will deliver a functional multioptrode array for large-volume patterned optogenetic stimulation. Parallel engineering efforts will add electrical recording capability, by utilizing the engineering resources already in place for the UEA, and will generate two types of integrated arrays. The “interleaved” array consists of an optrode array inserted through the back plane of a modified UEA into which a grid of through-backplane holes is made via laser ablation to accommodate the optrodes. For the “hybrid” array, each optrode shank will be coated with an isolation layer followed by a conductive layer, in order to allow recording while preventing light attenuation and stimulation artifacts. In vivo testing will assess the recording capabilities of both devices and subsequently the ability to perform simultaneous optical stimulation and electrical recordings. This technology will allow for unprecedented optogenetic investigations of mm-to-cm scale neural circuit function and dysfunction in NHPs, and for a new generation of therapeutic interventions via cell type specific optical neural control prosthetics.

了解非人类灵长类动物 (NHP) 大脑皮层神经回路的功能 最接近人类的模型系统，对于理解正常皮质功能和电路级基础至关重要 光遗传学已成为研究神经回路功能的强大工具。 利用光扰乱经过基因改造以表达光激活微生物的特定细胞类型的活性 视蛋白，并在成功的情况下评估这种扰动对网络活动和行为的影响。 在小鼠中，将光遗传学应用于 NHP 一直具有挑战性，这主要是由于缺乏多功能 集成探针可在毫米至厘米的体积内实现精确的光传输和电生理学 为了观察 NHP 皮层的深度，大容量操作对于大 NHP 大脑至关重要。 一个由 PI 组成的跨学科团队建议开发和测量可测量的电生理或行为效应。 测试体内集成穿透阵列，可实现大体积、时空图案化的光遗传学 NHP 大脑中神经回路的调制和电记录该项目需要协调。 4 个团队的努力，包括光子器件和光遗传学、材料和 µLED 开发方面的专家 生物相容性设备的包装、灵长类动物神经生理学以及电极阵列设计和领域的先驱 在目标 1 中，我们开发了该技术，在目标 2 中，我们在 NHP 视觉皮层中进行了体内测试。 我们最初将开发一个 4x4 mm 穿透性 10x10 光极阵列，其格式类似于井- 建立了犹他电阵列（UEA），每个探头充当波导，允许可见光到达 初步优化探头柄直径和尖端角度以最小化组织深度 >1.5mm。 组织损伤，我们将在深层皮质组织中进行体内 NHP 光遗传学实验的概念验证， 在第二阶段，我们将通过 µLED 开发光耦合， 它将被集成到一个平台中并进行体内测试，由位于每个平台上的 µLED 组成 第二阶段的完成将提供用于大体积图案化的功能性多光极阵列。 光遗传学刺激将通过利用光遗传学刺激来增加电记录能力。 UEA 的工程资源已经到位，并将生成两种类型的集成阵列。 “交错”阵列由一个光极阵列组成，该光极阵列插入修改后的 UEA 的背板，其中 通过激光烧蚀形成贯通背板孔的网格以容纳光极 对于“混合”阵列， 每个光极杆将涂有隔离层，然后是导电层，以便允许 记录的同时防止光衰减和刺激伪影 体内测试将评估记录。 两种设备的功能以及随后执行同时光刺激和 这项技术将允许对毫米到厘米进行前所未有的光遗传学研究。 衡量 NHP 的神经回路功能和功能障碍，以及新一代的治疗干预措施 通过细胞类型特定的光学神经控制假体。