Practical considerations in an era of multicolor optogenetics.

Practical considerations in an era of multicolor optogenetics.
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DOI:
10.3389/fncel.2023.1160245
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发表时间:
2023
影响因子:
5.3
通讯作者:
--
中科院分区:
医学2区
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--
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控制突触交流的能力是现代神经科学不可或缺的。直到最近,由于不同波长激活的视蛋白的可获得性有限,因此只能进行单路径操作。然而,广泛的蛋白质工程和筛选工作极大地扩展了光遗传工具箱,开启了一个研究神经回路的多色方法的时代。尽管如此,具有真正离散光谱的视蛋白仍然很少。因此,实验者必须小心避免光遗传工具的意外交叉激活(串扰)。在这里,我们通过测试刺激波长、辐照度、持续时间和视蛋白选择,展示了单个模型突触通路中串扰的多维性质。然后,我们提出了一种在逐个实验的基础上最大化光学响应的动态范围的“查找表”方法。
The ability to control synaptic communication is indispensable to modern neuroscience. Until recently, only single-pathway manipulations were possible due to limited availability of opsins activated by distinct wavelengths. However, extensive protein engineering and screening efforts have drastically expanded the optogenetic toolkit, ushering in an era of multicolor approaches for studying neural circuits. Nonetheless, opsins with truly discrete spectra are scarce. Experimenters must therefore take care to avoid unintended cross-activation of optogenetic tools (crosstalk). Here, we demonstrate the multidimensional nature of crosstalk in a single model synaptic pathway, testing stimulus wavelength, irradiance, duration, and opsin choice. We then propose a “lookup table” method for maximizing the dynamic range of opsin responses on an experiment-by-experiment basis.
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