In vivo optogenetic control of striatal and thalamic neurons in non-human primates.
In vivo optogenetic control of striatal and thalamic neurons in non-human primates.
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DOI：
10.1371/journal.pone.0050808
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Wichmann T
中科院分区：
文献类型：
--
作者：
Galvan A;Hu X;Smith Y;Wichmann T
电刺激和药物刺激方法通常用于研究体内神经元脑回路，但存在问题，因为电刺激具有有限的特异性，而药物激活具有低时间分辨率。最近开发的一种替代这些方法的方法是使用基于神经元中光敏通道蛋白表达的光遗传学技术。虽然光遗传学已经应用于啮齿动物的体外制备和体内研究，但它们在非人类灵长类动物脑功能研究中的应用仅限于大脑皮层。在这里，我们描述了通道视紫红质-2 （ChR2）转染在皮层下区域，即壳核、外白球（GPe）和腹侧丘脑（VL）的影响。在延伸因子1α启动子控制下，含有ChR2序列的慢病毒载体（plenti1 - ef1 α -hChR2(H134R)-eYFP-WPRE，滴度为109粒/ml）沉积在GPe、壳核和VL中。四周后，一个结合了传统电极和光纤的探针被引入先前注射的大脑区域。我们发现纹状体和丘脑中分别有31.5%和32.7%的神经元有光诱发反应，而GPe神经元只有2.5%有光诱发反应。正如预期的那样，大多数反应是时间锁定的放电增加，但也可以看到减少或混合反应，可能是通过chr2介导的局部抑制性连接的激活。光镜和电镜分析显示，ChR2在纹状体和VL的胞体、树突、棘和终末的质膜上强烈表达。本研究表明，在猴子身上成功地实现了针对纹状体和基底节区相关丘脑核的光遗传学实验。我们的研究结果表明，每种结构的响应类型和幅度存在重要差异。实验条件，如使用的载体，注射的数量和速度，或光刺激条件，必须优化每个结构的研究。
Electrical and pharmacological stimulation methods are commonly used to study neuronal brain circuits in vivo, but are problematic, because electrical stimulation has limited specificity, while pharmacological activation has low temporal resolution. A recently developed alternative to these methods is the use of optogenetic techniques, based on the expression of light sensitive channel proteins in neurons. While optogenetics have been applied in in vitro preparations and in in vivo studies in rodents, their use to study brain function in nonhuman primates has been limited to the cerebral cortex. Here, we characterize the effects of channelrhodopsin-2 (ChR2) transfection in subcortical areas, i.e., the putamen, the external globus pallidus (GPe) and the ventrolateral thalamus (VL) of rhesus monkeys. Lentiviral vectors containing the ChR2 sequence under control of the elongation factor 1α promoter (pLenti-EF1α -hChR2(H134R)-eYFP-WPRE, titer 109 particles/ml) were deposited in GPe, putamen and VL. Four weeks later, a probe combining a conventional electrode and an optic fiber was introduced in the previously injected brain areas. We found light-evoked responses in 31.5% and 32.7% of all recorded neurons in the striatum and thalamus, respectively, but only in 2.5% of recorded GPe neurons. As expected, most responses were time-locked increases in firing, but decreases or mixed responses were also seen, presumably via ChR2-mediated activation of local inhibitory connections. Light and electron microscopic analyses revealed robust expression of ChR2 on the plasma membrane of cell somas, dendrites, spines and terminals in the striatum and VL. This study demonstrates that optogenetic experiments targeting the striatum and basal ganglia-related thalamic nuclei can be successfully achieved in monkeys. Our results indicate important differences of the type and magnitude of responses in each structure. Experimental conditions such as the vector used, the number and rate of injections, or the light stimulation conditions have to be optimized for each structure studied.
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纹状体培养基神经元的功能连接组。
DOI：
10.1523/jneurosci.3833-10.2011
发表时间：
2011-01-26
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Chuhma N;Tanaka KF;Hen R;Rayport S
通讯作者：
Rayport S
使用ChannelRhopopsin-2的细胞型特异性表达，对体内神经元的靶向光遗传学刺激和记录。
影响因子：
14.8
作者：
通讯作者：
--
影响因子：
25
作者：
Boyden, ES;Zhang, F;Deisseroth, K
通讯作者：
Deisseroth, K
影响因子：
2.5
作者：
Galvan, A;Villalba, RM;Wichmann, T
通讯作者：
Wichmann, T
影响因子：
19.9
作者：
Bernstein, Jacob G.;Boyden, Edward S.
通讯作者：
Boyden, Edward S.