Hybrid voltage sensor imaging of electrical activity from neurons in hippocampal slices from transgenic mice.
Hybrid voltage sensor imaging of electrical activity from neurons in hippocampal slices from transgenic mice.
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DOI：
10.1152/jn.00722.2012
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发表时间：
2012-12
影响因子：
2.5
通讯作者：
Dongsheng Wang;S. McMahon;Zhen Zhang;M. Jackson
中科院分区：
文献类型：
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作者：
Dongsheng Wang;S. McMahon;Zhen Zhang;M. Jackson
基因靶向与遗传编码的光学电压传感器带来了电压成像的方法，遗传定义的神经元，并提供了一种方法，研究电路活动在这些选定的人群。本研究报告的遗传编码的混合电压传感器（hVOS）的转基因小鼠的神经元的目标。hVOS系列探针采用膜靶向荧光蛋白，其在二苦胺（DPA）存在下产生电压依赖性荧光变化，这是两个分子之间的电压依赖性光学相互作用的结果。我们用两种不同的高性能hVOS探针在神经元特异性thy-1启动子的控制下产生转基因小鼠。这些动物的海马切片呈现不同的表达空间模式，电刺激诱发的荧光变化高达3%。谷氨酸受体和Na（+）通道拮抗剂阻断了这些反应。这里测试的一个hVOS探针含有轴突靶向基序（来自GAP-43），并显示在轴突中的优先表达;因此，该探针可以报告轴突电压变化。表达hVOS探针的转基因小鼠的电压成像为研究完整神经回路中遗传定义的神经元群体的功能活动打开了大门。
Gene targeting with genetically encoded optical voltage sensors brings the methods of voltage imaging to genetically defined neurons and offers a method of studying circuit activity in these selected populations. The present study reports the targeting of genetically encoded hybrid voltage sensors (hVOS) to neurons in transgenic mice. The hVOS family of probes employs a membrane-targeted fluorescent protein, which generates voltage-dependent fluorescence changes in the presence of dipicrylamine (DPA) as the result of a voltage-dependent optical interaction between the two molecules. We generated transgenic mice with two different high-performance hVOS probes under control of a neuron-specific thy-1 promoter. Hippocampal slices from these animals present distinct spatial patterns of expression, and electrical stimulation evoked fluorescence changes as high as 3%. Glutamate receptor and Na(+) channel antagonists blocked these responses. One hVOS probe tested here harbors an axonal targeting motif (from GAP-43) and shows preferential expression in axons; this probe can thus report axonal voltage changes. Voltage imaging in transgenic mice expressing hVOS probes opens the door to the study of functional activity in genetically defined populations of neurons in intact neural circuits.