Off-Peak 594-nm Light Surpasses On-Peak 532-nm Light in Silencing Distant ArchT-Expressing Neurons In Vivo
Off-Peak 594-nm Light Surpasses On-Peak 532-nm Light in Silencing Distant ArchT-Expressing Neurons In Vivo
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DOI：
10.1016/j.isci.2020.101276
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发表时间：
2020-07-24
期刊：
影响因子：
5.8
通讯作者：
Miyashita，Yasushi
中科院分区：
文献类型：
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作者：
Setsuie，Rieko;Tamura，Keita;Miyashita，Yasushi
对于使用光遗传学的大脑体积操作，需要有效的视蛋白激发和具有最小光吸收的高效光传输，以最大限度地减少照明光强度和伴随的脱靶效应。 ArchT 是一种广泛使用的强效抑制性视蛋白，通常由 532 nm 光激活，该光位于其体外激发峰。然而，532 nm 光也位于血红蛋白吸收光谱的峰值范围内。因此，我们预测 594 nm 光在抑制远处表达 ArchT 的神经元方面具有优越性，该神经元稍微偏离 ArchT 激发平台，并且很大程度上偏离血红蛋白吸收光谱的峰值。我们通过大鼠皮质蛋白体内电生理记录定量测试了这一预测。当照明距离大于 500 μm 时，594 nm 光比 532 nm 光更有效。它的优势随着距离的增加而增加。这些结果验证了我们的预测，并强调了激发-吸收权衡在选择体内光遗传学照明波长时的重要性。
For large brain volume manipulations using optogenetics, both effective opsin excitation and efficient light delivery with minimal light absorption are required to minimize the illuminating light intensity and concomitant off-target effects. ArchT, a widely used potent inhibitory opsin, is commonly activated by 532-nm light, which lies on itsin vitroexcitation peak. However, 532-nm light also lies on a peak range of the hemoglobin absorption spectrum. Therefore, we predicted that 594-nm light is superior in suppressing distant ArchT-expressing neurons, which is slightly off the ArchT-excitation-plateau and largely off the peak of the hemoglobin absorption spectrum. We quantitatively tested this prediction by the electrophysiological recording of the rat cortexin vivo. At illumination distances greater than 500 μm, 594-nm light was more effective than 532-nm light. Its superiority increased with distance. These results validate our prediction and highlight the significance of excitation-absorption trade-off in selecting illumination wavelength for optogeneticsin vivo.