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首页> 外文期刊>Neuron >Optogenetic inhibition of synaptic release with chromophore-assisted light inactivation (CALI)
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Optogenetic inhibition of synaptic release with chromophore-assisted light inactivation (CALI)

机译：生色团辅助光灭活（CALI）对突触释放的光遗传学抑制

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摘要

Optogenetic techniques provide effective ways of manipulating the functions of selected neurons with light. In the current study, we engineered an optogenetic technique that directly inhibits neurotransmitter release. We used a genetically encoded singlet oxygen generator, miniSOG, to conduct chromophore assisted light inactivation (CALI) of synaptic proteins. Fusions of miniSOG to VAMP2 and synaptophysin enabled disruption of presynaptic vesicular release upon illumination with blue light. In cultured neurons and hippocampal organotypic slices, synaptic release was reduced up to 100%. Such inhibition lasted >1hr and had minimal effects on membrane electrical properties. When miniSOG-VAMP2 was expressed panneuronally in Caenorhabditis elegans, movement of the worms was reduced after illumination, and paralysis was often observed. The movement of the worms recovered overnight. We name this technique Inhibition of Synapses with CALI (InSynC). InSynC is a powerful way to silence genetically specified synapses with light in a spatially and temporally precise manner.
机译：光遗传学技术提供了利用光操纵选定神经元功能的有效方法。在当前的研究中，我们设计了一种直接抑制神经递质释放的光遗传学技术。我们使用遗传编码的单线态氧发生器miniSOG来进行突触蛋白的生色团辅助光灭活（CALI）。 miniSOG与VAMP2和突触素的融合能够在蓝光照射下破坏突触前囊泡的释放。在培养的神经元和海马器官型切片中，突触释放最多可降低100％。这种抑制作用持续＞ 1小时，并且对膜的电性能影响最小。当miniSOG-VAMP2在秀丽隐杆线虫泛神经元表达时，蠕虫的移动在光照后减少，并且经常观察到瘫痪。蠕虫的移动恢复了一夜。我们将这种技术命名为CALI抑制突触（InSynC）。 InSynC是一种以时空精确的方式用光沉默遗传学突触的有效方法。

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