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首页> 外文期刊>Brain cell biology >Optimizing the spatial resolution of Channelrhodopsin-2 activation.
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Optimizing the spatial resolution of Channelrhodopsin-2 activation.

机译：优化Channelrhodopsin-2激活的空间分辨率。

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摘要

Over the past few years, the light-gated cation channel Channelrhodopsin-2 (ChR2) has seen a remarkable diversity of applications in neuroscience. However, commonly used wide-field illumination provides poor spatial selectivity for cell stimulation. We explored the potential of focal laser illumination to map photocurrents of individual neurons in sparsely transfected hippocampal slice cultures. Interestingly, the best spatial resolution of photocurrent induction was obtained at the lowest laser power. By adjusting the light intensity to a neuron's spike threshold, we were able to trigger action potentials with a spatial selectivity of less than 30 microm. Experiments with dissociated hippocampal cells suggested that the main factor limiting the spatial resolution was ChR2 current density rather than scattering of the excitation light. We conclude that subcellular resolution can be achieved only in cells with a high ChR2 expression level and that future improved variants of ChR2 are likely to extend the spatial resolution of photocurrent induction to the level of single dendrites.
机译：在过去的几年中，光门控阳离子通道Channelrhodopsin-2（ChR2）在神经科学中已获得了广泛的应用。但是，通常使用的广角照明对细胞刺激的空间选择性差。我们探索了在稀疏转染的海马切片培养物中聚焦激光照明来绘制单个神经元光电流的潜力。有趣的是，在最低的激光功率下获得了最佳的光电流感应空间分辨率。通过将光强度调整到神经元的尖峰阈值，我们能够以小于30微米的空间选择性触发动作电位。分离海马细胞的实验表明，限制空间分辨率的主要因素是ChR2电流密度，而不是激发光的散射。我们得出的结论是，只有在具有高ChR2表达水平的细胞中才能实现亚细胞分辨率，并且未来改进的ChR2变体可能会将光电流诱导的空间分辨率扩展到单个树突的水平。

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