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非人类灵长类动物中光阵列植入的手术程序。

Surgical Procedure for Implantation of Opto-Array in Nonhuman Primates.

机构信息

Laboratory of Neuropsychology, National Institute of Mental Health, NIH, Bethesda, Maryland.

Department of Psychology, University of Pennsylvania, Philadelphia, Pennsylvania.

出版信息

Curr Protoc. 2023 Mar;3(3):e704. doi: 10.1002/cpz1.704.

Abstract

Optogenetics allows precise temporal control of neuronal activity in the brain. Engineered viral vectors are routinely used to transduce neurons with light-sensitive opsins. However, reliable virus injection and light delivery in animals with large brains, such as nonhuman primates, has proven challenging. The Opto-Array is a novel yet simple device that is used to deliver light to extended regions of the cortex surface for high-throughput behavioral optogenetics in large brains. Here we present protocols for surgical delivery of virus (Basic Protocol 1) and implantation of the Opto-Array (Basic Protocol 2) in two separate surgeries in a rhesus monkey's inferior temporal cortex. As a proof of concept, we measured the behavioral performance of an animal detecting cortical optogenetic stimulations (Basic Protocol 3) with different illumination power and duration using the Opto-Array. The animal was able to detect the optogenetic stimulation for all tested illumination powers and durations. Regression analysis also showed both power and duration of illumination significantly modulate the detectability of the optogenetic stimulation. The outcome of this approach is superior to the standard practice of injecting and recording through a chamber for large areas of the cortex surface. Moreover, the chronic nature of the Opto-Array allows perturbation of neuronal activity of the same site across multiple sessions because it is highly stable; thus, data can be pooled over months. The detailed surgical method presented here makes it possible to use optogenetics to modulate neuronal activity across large regions of the cortex surface in the nonhuman primate brain. This method also lays the groundwork for future attempts to use optogenetics to restore vision in humans. © 2023 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Virus injection surgery Basic Protocol 2: Opto-Array implantation surgery Basic Protocol 3: Cortical Perturbation Detection (CPD) task behavioral training.

摘要

光遗传学允许在大脑中精确控制神经元的活动。工程病毒载体通常用于转导具有光敏感视蛋白的神经元。然而,在大型动物(如非人类灵长类动物)中,可靠的病毒注射和光传递一直是一个挑战。Opto-Array 是一种新颖而简单的设备,用于向皮质表面的扩展区域提供光,以在大型大脑中进行高通量行为光遗传学研究。在此,我们介绍了在恒河猴颞叶皮质中通过两次手术分别进行病毒(基本方案 1)和 Opto-Array 植入(基本方案 2)的方案。作为概念验证,我们使用 Opto-Array 测量了一只动物检测皮质光遗传学刺激的行为表现(基本方案 3),其中包括不同的光照功率和持续时间。动物能够检测到所有测试的光照功率和持续时间的光遗传学刺激。回归分析还表明,光照的功率和持续时间都显著调节了光遗传学刺激的可检测性。该方法的结果优于在皮质表面大面积上通过腔室注射和记录的标准做法。此外,Opto-Array 的慢性性质允许在多个会话中对同一部位的神经元活动进行干扰,因为它非常稳定;因此,数据可以在数月内汇总。本文介绍的详细手术方法使得在非人类灵长类动物大脑中通过光遗传学调节皮质表面大片区的神经元活动成为可能。该方法也为未来使用光遗传学恢复人类视力的尝试奠定了基础。© 2023 作者。 Wiley Periodicals LLC 出版的《当代协议》。 基本方案 1:病毒注射手术 基本方案 2:Opto-Array 植入手术 基本方案 3:皮质干扰检测(CPD)任务行为训练。

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