Department of Complexity Science and Engineering, University of Tokyo, Kashiwanoha, Chiba, Japan.
Department of Physics, Graduate School of Science, University of Tokyo, Bunkyo-ku, Tokyo, Japan.
Adv Exp Med Biol. 2021;1293:309-320. doi: 10.1007/978-981-15-8763-4_19.
The fruit fly Drosophila melanogaster, an insect 4 mm long, has served as the experimental subject in a wide range of biological research, including neuroscience. In this chapter, we briefly introduce optogenetic applications in Drosophila neuroscience research. First, we describe the development of Drosophila from egg to adult. In fly neuroscience, temperature-controlled perturbation of neural activity, sometimes called "thermogenetics," has been an invaluable tool that predates the advent of optogenetics. After briefly introducing this perturbation technique, we describe the process of generating transgenic flies that express optogenetic probes in a specific group of cells. Transgenic techniques are crucial in the application of optogenetics in Drosophila neuroscience; here we introduce the transposon P-elements, ϕC31 integrase, and CRISPR-Cas9 methods. As for cell-specific gene expression techniques, the binary expression systems utilizing Gal4-UAS, LexA-lexAop, and Q-system are described. We also present a short and basic optogenetic experiment with Drosophila larvae as a practical example. Finally, we review a few recent studies in Drosophila neuroscience that made use of optogenetics. In this overview of fly development, transgenic methods, and applications of optogenetics, we present an introductory background to optogenetics in Drosophila.
果蝇（Drosophila melanogaster）是一种 4 毫米长的昆虫，已被广泛应用于生物学研究，包括神经科学。在本章中，我们将简要介绍光遗传学在果蝇神经科学研究中的应用。首先，我们描述了果蝇从卵到成虫的发育过程。在果蝇神经科学中，对神经活动进行温度控制的干扰，有时被称为“热遗传学”，是在光遗传学出现之前就已经非常有用的工具。在简要介绍了这种干扰技术之后，我们描述了生成在特定细胞群中表达光遗传学探针的转基因果蝇的过程。转基因技术是光遗传学在果蝇神经科学中的应用的关键；在这里，我们介绍转座子 P 元件、ϕC31 整合酶和 CRISPR-Cas9 方法。至于细胞特异性基因表达技术，我们描述了利用 Gal4-UAS、LexA-lexAop 和 Q 系统的二元表达系统。我们还提供了一个使用果蝇幼虫的简单基本光遗传学实验作为实际示例。最后，我们回顾了一些最近利用光遗传学进行的果蝇神经科学研究。通过对果蝇发育、转基因方法和光遗传学应用的概述，我们为光遗传学在果蝇中的应用提供了一个入门背景。
