Won Joungha, Pankratov Yuriy, Jang Minwoo Wendy, Kim Sunpil, Ju Yeon Ha, Lee Sangkyu, Lee Seung Eun, Kim Arie, Park Soowon, Lee C Justin, Heo Won Do
Center for Cognition and Sociality, Institute for Basic Science (IBS), Daejeon 34126, Republic of Korea; Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon 34141, Republic of Korea.
School of Life Sciences, University of Warwick, Coventry, CV4 7AL, UK.
Neuron. 2022 Feb 2;110(3):423-435.e4. doi: 10.1016/j.neuron.2021.11.003. Epub 2021 Nov 30.
Spatiotemporal control of brain activity by optogenetics has emerged as an essential tool to study brain function. For silencing brain activity, optogenetic probes, such as halorhodopsin and archaerhodopsin, inhibit transmitter release indirectly by hyperpolarizing membrane potentials. However, these probes cause an undesirable ionic imbalance and rebound spikes. Moreover, they are not applicable to use in non-excitable glial cells. Here we engineered Opto-vTrap, a light-inducible and reversible inhibition system to temporarily trap the transmitter-containing vesicles from exocytotic release. Light activation of Opto-vTrap caused full vesicle clusterization and complete inhibition of exocytosis within 1 min, which recovered within 30 min after light off. We found a significant reduction in synaptic and gliotransmission upon activation of Opto-vTrap in acute brain slices. Opto-vTrap significantly inhibited hippocampus-dependent memory retrieval with full recovery within an hour. We propose Opto-vTrap as a next-generation optogenetic silencer to control brain activity and behavior with minimal confounding effects.
光遗传学对大脑活动的时空控制已成为研究大脑功能的重要工具。对于沉默大脑活动，诸如盐视紫红质和古菌视紫红质等光遗传学探针通过使膜电位超极化来间接抑制神经递质释放。然而，这些探针会导致不良的离子失衡和反弹尖峰。此外，它们不适用于非兴奋性神经胶质细胞。在此，我们构建了Opto-vTrap，这是一种光诱导且可逆的抑制系统，用于暂时捕获含神经递质的囊泡使其无法进行胞吐释放。Opto-vTrap的光激活在1分钟内导致囊泡完全聚集并完全抑制胞吐作用，在光照关闭后30分钟内恢复。我们发现在急性脑片中激活Opto-vTrap后，突触传递和胶质传递显著减少。Opto-vTrap显著抑制海马体依赖的记忆提取，且在一小时内完全恢复。我们提出Opto-vTrap作为下一代光遗传学沉默工具，以在最小化混杂效应的情况下控制大脑活动和行为。
