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.

了解神经回路在非人类灵长类动物（NHP）的大脑皮层中的功能， 最接近人类的模型系统对于理解正常皮质功能和电路级别至关重要 人脑疾病。光遗传学已成为研究神经电路功能的强大工具， 使用光扰动特定细胞类型的活性，通常修改以表达光激活微生物 Opsins，并评估这种扰动对网络活动和行为的后果。虽然成功 在小鼠中，将光遗传学应用于NHP的挑战很大，这主要是由于缺乏多功能 综合问题，以通过MM到CM的精确光输送和电生理学通过 NHP皮层的深度。在大型NHP大脑中，大容量的操作至关重要，以观察 可测量的电生理或行为效应。 PIS的跨学科团队，要开发和 在体内综合渗透阵列中测试，该阵列允许大量，时空图案化的光遗传学 NHP脑中神经元电路的调节和电记录。该项目需要协调 4个团队的努力，包括光子设备方面的专家以及用于光遗传学，材料和 用于生物相容性设备，一级神经生理学和电极阵列设计中的先驱者的包装 商业化。在AIM 1中，我们开发了该技术，在AIM 2中，我们在NHP Visual Cortex中的体内对其进行了测试。 我们最初将以类似于井的格式开发4x4 mm穿透10x10 optrode阵列 已建立的犹他州电动阵列（UEA），每个探针用作波导，允许可见光到达 组织深度> 1.5mm。最初优化了探针的柄直径和尖端角度以最小化 组织损伤，我们将在深层皮质组织中进行体内NHP光学遗传实验的概念验证， 使用整个阵列的宽区域照明。在第二阶段，我们将通过µled开发光耦合， 它将集成到一个平台中并在体内进行测试，由每个位于每个平台 光学探针。第2阶段的完成将提供一个功能性的多流动阵列，用于大容量的图案 光遗传学刺激。并行工程工作将通过使用 UEA已经有了工程资源，并将生成两种类型的集成阵列。这 “交错”阵列由插入修改后的UEA的背面插入的Optrode阵列组成， 通过激光消融制成透明孔孔的网格以容纳开孔。对于“混合”阵列， 每个Optrode柄将用一个隔离层覆盖，然后是导电层，以便允许 记录同时防止光衰减和刺激伪像。体内测试将评估记录 设备的功能以及随后执行同时光学刺激的能力和 电气记录。该技术将允许对MM到CM进行前所未有的光遗传学研究 NHP中的缩放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肢。