Non-invasive Strategies for Chronic Manipulation of DREADD-controlled Neuronal Activity.
Non-invasive Strategies for Chronic Manipulation of DREADD-controlled Neuronal Activity.
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DOI：
10.3791/59439
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发表时间：
2019-08
期刊：
影响因子：
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通讯作者：
Jesse J. Zhan;Ruchi Komal;W. T. Keenan;S. Hattar;D. C. Fernandez
中科院分区：
文献类型：
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作者：
Jesse J. Zhan;Ruchi Komal;W. T. Keenan;S. Hattar;D. C. Fernandez
化学遗传学策略已成为远程控制神经元活动的可靠工具。其中，仅由设计药物激活的设计受体（DREADD）已成为现代神经科学中最流行的化学遗传学方法。大多数研究通过单次腹腔注射提供配体氯氮平-N-氧化物（CNO），这适用于目标神经元群的急性激活/抑制。然而，慢性调节 DREADD 控制的神经元的策略例子很少，其中大多数依赖于使用需要手术干预的输送系统。在这里，我们扩展了两种非侵入性策略，用于递送配体 CNO 以长期操纵小鼠的神经群。 CNO 通过使用重复（每日）滴眼剂或通过动物的饮用水长期施用。这些非侵入性范例导致设计受体的强烈激活，并在整个 CNO 治疗过程中持续存在。这里描述的方法为慢性 DREADD 介导的神经元活动控制提供了替代方案，并且可能适用于旨在评估自由活动动物行为的实验，重点关注侵入性较小的 CNO 传递方法。
Chemogenetic strategies have emerged as reliable tools for remote control of neuronal activity. Among these, designer receptors exclusively activated by designer drugs (DREADDs) have become the most popular chemogenetic approach used in modern neuroscience. Most studies deliver the ligand clozapine-N-oxide (CNO) using a single intraperitoneal injection, which is suitable for the acute activation/inhibition of the targeted neuronal population. There are, however, only a few examples of strategies for chronic modulation of DREADD-controlled neurons, the majority of which rely on the use of delivery systems that require surgical intervention. Here, we expand on two non-invasive strategies for delivering the ligand CNO to chronically manipulate neural population in mice. CNO was administered either by using repetitive (daily) eye-drops, or chronically through the animal's drinking water. These non-invasive paradigms result in robust activation of the designer receptors that persisted throughout the CNO treatments. The methods described here offer alternatives for the chronic DREADD-mediated control of neuronal activity and may be useful for experiments designed to evaluate behavior in freely moving animals, focusing on less-invasive CNO delivery methods.