Hsiao Po-Yen, Wu Ming-Chin, Lin Yen-Yin, Fu Chein-Chung, Chiang Ann-Shyn
Institute of Biotechnology, National Tsing Hua University, Hsinchu, 30013, Taiwan.
Brain Research Center, National Tsing Hua University, Hsinchu, 30013, Taiwan.
Methods Mol Biol. 2016;1408:377-87. doi: 10.1007/978-1-4939-3512-3_26.
Activating selected neurons elicits specific behaviors in Drosophila adults. By combining optogenetics and laser-tracking techniques, we have recently developed an automated laser-tracking and optogenetic manipulation system (ALTOMS) for studying how brain circuits orchestrate complex behaviors. The established ALTOMS can independently target three lasers (473-nm blue laser, 593.5-nm yellow laser, and 1064-nm infrared laser) on any specified body part of two freely moving flies. Triggering light-sensitive proteins in real time, the blue laser and yellow laser can respectively activate and inhibit target neurons in artificial transgenic flies. Since infrared light is invisible to flies, we use the 1064-nm laser as an aversive stimulus in operant learning without perturbing visual inputs. Herein, we provide a detailed protocol for the construction of ALTOMS and optogenetic manipulation of target neurons in Drosophila adults during social interactions.
激活选定的神经元会引发果蝇成虫的特定行为。通过结合光遗传学和激光追踪技术，我们最近开发了一种自动激光追踪和光遗传学操纵系统（ALTOMS），用于研究脑回路如何协调复杂行为。已建立的ALTOMS可以将三台激光器（473纳米蓝光激光器、593.5纳米黄光激光器和1064纳米红外激光器）独立瞄准两只自由飞行果蝇的任何指定身体部位。蓝光激光器和黄光激光器实时触发光敏感蛋白，可分别激活和抑制人工转基因果蝇中的靶神经元。由于果蝇对红外光不可见，我们使用1064纳米激光器作为操作性学习中的厌恶刺激，而不会干扰视觉输入。在此，我们提供了一份详细的方案，用于构建ALTOMS以及在社交互动期间对果蝇成虫的靶神经元进行光遗传学操纵。
