Fiber-optic implantation for chronic optogenetic stimulation of brain tissue.
Fiber-optic implantation for chronic optogenetic stimulation of brain tissue.
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DOI：
10.3791/50004
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发表时间：
2012-10-29
期刊：
影响因子：
--
通讯作者：
Arenkiel BR
中科院分区：
文献类型：
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作者：
Ung K;Arenkiel BR
光遗传学的发展现在提供了在体外和体内精确刺激基因定义的神经元和电路的手段。在这里，我们描述的组装和植入光纤用于慢性光刺激脑组织。阐明神经元连接的模式对临床和基础神经科学都是一个挑战。电生理学一直是分析突触连接模式的黄金标准，但是成对的电生理学记录既繁琐又在实验上受到限制。光遗传学的发展引入了一种优雅的方法来刺激神经元和电路，无论是在体外还是在体内。通过利用细胞类型特异性启动子活性来驱动离散神经元群体中的视蛋白表达，可以精确地刺激不同回路中遗传定义的神经元亚型。刺激神经元的方法，包括电刺激和/或药理学操作，通常是不区分细胞类型的，侵入性的，并且可能损害周围组织。这些限制可能会改变正常的突触功能和/或电路行为。此外，由于手法的性质，目前的方法往往是急性和终末的。光遗传学提供了以相对无害的方式刺激神经元的能力，并在基因上靶向神经元。目前，大多数涉及体内光遗传学的研究都使用通过植入导管引导的光纤；然而，这种方法的局限性包括反复插入光纤会损伤脑组织，以及可能破坏套管内的纤维。鉴于光遗传学领域的蓬勃发展，需要一种更可靠的慢性刺激方法来促进长期研究，同时将附带组织损伤降到最低。在这里，我们将我们的修改方案作为视频文章提供，以有效而优雅地补充Sparta等人描述的方法，用于制造光纤植入物并将其永久固定在麻醉小鼠的头骨上，以及将植入物连接到光源的光纤耦合器的组装。这种植入物用光纤连接到固态激光器，使用小的、可拆卸的系绳，可以有效地长期光刺激功能性神经回路，同时减少组织损伤。光纤植入物的永久固定为清醒、行为正常的小鼠提供了一致的、长期的体内光遗传学研究。
The development of optogenetics now provides the means to precisely stimulate genetically defined neurons and circuits, both in vitro and in vivo. Here we describe the assembly and implantation of a fiber optic for chronic photostimulation of brain tissue. Elucidating patterns of neuronal connectivity has been a challenge for both clinical and basic neuroscience. Electrophysiology has been the gold standard for analyzing patterns of synaptic connectivity, but paired electrophysiological recordings can be both cumbersome and experimentally limiting. The development of optogenetics has introduced an elegant method to stimulate neurons and circuits, both in vitro and in vivo. By exploiting cell-type specific promoter activity to drive opsin expression in discrete neuronal populations, one can precisely stimulate genetically defined neuronal subtypes in distinct circuits. Well described methods to stimulate neurons, including electrical stimulation and/or pharmacological manipulations, are often cell-type indiscriminate, invasive, and can damage surrounding tissues. These limitations could alter normal synaptic function and/or circuit behavior. In addition, due to the nature of the manipulation, the current methods are often acute and terminal. Optogenetics affords the ability to stimulate neurons in a relatively innocuous manner, and in genetically targeted neurons. The majority of studies involving in vivo optogenetics currently use a optical fiber guided through an implanted cannula; however, limitations of this method include damaged brain tissue with repeated insertion of an optical fiber, and potential breakage of the fiber inside the cannula. Given the burgeoning field of optogenetics, a more reliable method of chronic stimulation is necessary to facilitate long-term studies with minimal collateral tissue damage. Here we provide our modified protocol as a video article to complement the method effectively and elegantly described in Sparta et al. for the fabrication of a fiber optic implant and its permanent fixation onto the cranium of anesthetized mice, as well as the assembly of the fiber optic coupler connecting the implant to a light source. The implant, connected with optical fibers to a solid-state laser, allows for an efficient method to chronically photostimulate functional neuronal circuitry with less tissue damage using small, detachable, tethers. Permanent fixation of the fiber optic implants provides consistent, long-term in vivo optogenetic studies of neuronal circuits in awake, behaving mice with minimal tissue damage.
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影响因子：
14.8
作者：
Zhang F;Gradinaru V;Adamantidis AR;Durand R;Airan RD;de Lecea L;Deisseroth K
通讯作者：
Deisseroth K
影响因子：
16.2
作者：
Arenkiel, Benjamin R.;Peca, Joao;Feng, Guoping
通讯作者：
Feng, Guoping
影响因子：
25
作者：
Boyden, ES;Zhang, F;Deisseroth, K
通讯作者：
Deisseroth, K
影响因子：
64.8
作者：
Adamantidis, Antoine R.;Zhang, Feng;De Lecea, Luis
通讯作者：
De Lecea, Luis
影响因子：
64.8
作者：
通讯作者：
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