Remote and reversible inhibition of neurons and circuits by small molecule induced potassium channel stabilization.
Remote and reversible inhibition of neurons and circuits by small molecule induced potassium channel stabilization.
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通过小分子诱导钾通道稳定对神经元和电路进行远程和可逆抑制。
DOI：
10.1038/srep19293
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发表时间：
2016-01-13
影响因子：
4.6
通讯作者：
Thoeringer CK
中科院分区：
文献类型：
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作者：
Auffenberg E;Jurik A;Mattusch C;Stoffel R;Genewsky A;Namendorf C;Schmid RM;Rammes G;Biel M;Uhr M;Moosmang S;Michalakis S;Wotjak CT;Thoeringer CK
操纵神经元和电路的功能，将电信号和化学信号转化为行为，是神经科学的一个主要挑战。除了使用光激活通道的光遗传学方法外，还开发了配体诱导细胞信号传导和兴奋性调节的药物遗传学方法。然而，它们主要是基于外源或嵌合体蛋白的异位表达。现在，我们利用小分子诱导蛋白稳定的化学发生技术描述了Kir2.1型钾通道的远程和可逆表达。基于屏蔽1介导的不稳定结构域与感兴趣的蛋白质融合的脱落和蛋白质降解的抑制，这一原则已被用于生物医学，但迄今尚未在神经科学中采用。在这里，我们首次将这种化学遗传学方法应用于大脑研究，以便以远程和可逆的方式控制钾通道。我们可以证明，在体外和体内，屏蔽1介导的异位Kir2.1稳定诱导了小鼠大脑中的神经元沉默。我们还在不同的神经行为范例中验证了这种新的药物遗传方法。DD-Kir2.1可以补充现有的药物和光遗传学技术组合，用于特定的神经元操作，但它也可以为该原理在神经科学研究中的未来应用提供一个例子。
Manipulating the function of neurons and circuits that translate electrical and chemical signals into behavior represents a major challenges in neuroscience. In addition to optogenetic methods using light-activatable channels, pharmacogenetic methods with ligand induced modulation of cell signaling and excitability have been developed. However, they are largely based on ectopic expression of exogenous or chimera proteins. Now, we describe the remote and reversible expression of a Kir2.1 type potassium channel using the chemogenetic technique of small molecule induced protein stabilization. Based on shield1-mediated shedding of a destabilizing domain fused to a protein of interest and inhibition of protein degradation, this principle has been adopted for biomedicine, but not in neuroscience so far. Here, we apply this chemogenetic approach in brain research for the first time in order to control a potassium channel in a remote and reversible manner. We could show that shield1-mediated ectopic Kir2.1 stabilization induces neuronal silencing in vitro and in vivo in the mouse brain. We also validated this novel pharmacogenetic method in different neurobehavioral paradigms.The DD-Kir2.1 may complement the existing portfolio of pharmaco- and optogenetic techniques for specific neuron manipulation, but it may also provide an example for future applications of this principle in neuroscience research.