Using Drosophila Two-Choice Assay to Study Optogenetics in Hands-On Neurobiology Laboratory Activities.

Zhuo Fu, Ainul Huda, Ian F Kimbrough, Lina Ni
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Abstract

Optogenetics has made a significant impact on neuroscience, allowing activation and inhibition of neural activity with exquisite spatiotemporal precision in response to light. In this lab session, we use fruit flies to help students understand the fundamentals of optogenetics through hands-on activities. The CsChrimson channelrhodopsin, a light-activated cation channel, is expressed in sweet and bitter sensory neurons. Sweet sensory neurons guide animals to identify nutrient-rich food and drive appetitive behaviors, while bitter sensory neurons direct animals to avoid potentially toxic substances and guide aversive behavior. Students use two-choice assays to explore the causality between the stimulation activation of these neurons and the appetitive and avoidance behaviors of the fruit flies. To quantify their observations, students calculate preference indices and use the Student's t-test to analyze their data. After this lab session, students are expected to have a basic understanding of optogenetics, fly genetics, sensory perception, and how these relate to sensory-guided behaviors. They will also learn to conduct, quantify, and analyze two-choice behavioral assays.

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在动手神经生物学实验室活动中使用果蝇双选法研究光遗传学。
光遗传学对神经科学产生了重大影响,它可以在光的作用下,以精妙的时空精度激活和抑制神经活动。在这节实验课中,我们利用果蝇,通过实践活动帮助学生了解光遗传学的基本原理。CsChrimson 通道闪烁蛋白是一种光激活阳离子通道,在甜味和苦味感觉神经元中均有表达。甜味感觉神经元引导动物识别营养丰富的食物并驱动食欲行为,而苦味感觉神经元则引导动物避开潜在的有毒物质并引导厌恶行为。学生们利用双项选择实验来探索这些神经元的刺激激活与果蝇的食欲和回避行为之间的因果关系。为了量化观察结果,学生会计算偏好指数,并使用学生 t 检验分析数据。完成本实验课后,学生将对光遗传学、果蝇遗传学、感官知觉以及它们与感官引导行为的关系有基本的了解。他们还将学会进行、量化和分析双选行为实验。
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