Developing Drosophila melanogaster Models for Imaging and Optogenetic Control of Cardiac Function.
Developing Drosophila melanogaster Models for Imaging and Optogenetic Control of Cardiac Function.
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DOI：
10.3791/63939
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发表时间：
2022-08-25
期刊：
影响因子：
--
通讯作者：
Zhou C
中科院分区：
文献类型：
--
作者：
Gracheva E;Wang F;Matt A;Liang H;Fishman M;Zhou C
利用果蝇(果蝇)作为模式生物，在生物科学的许多领域，从细胞组织、基因组研究到行为研究，都取得了重大进展。由于积累了科学知识，近年来，果蝇被带到了包括心脏病在内的人类疾病建模领域。目前的工作描述了一种实验系统，用于在整个活体的背景下使用红光(617 Nm)并且没有侵入性程序来监测和操纵心脏功能。对心脏的控制是使用光遗传工具实现的。光遗传学结合了光敏转基因opsins的表达和它们的光激活来调节感兴趣的生物组织。在这项工作中，定制的集成光学相干断层扫描(OCT)成像和光遗传刺激系统被用来可视化和调节3龄幼虫和早期蛹发育阶段的黑腹果蝇心脏的功能。利用UAS/GAL4双基因系统在果蝇心脏特异表达卤视紫红质(eNpHR2.0)和红移通道视紫红质(ReaChR)。提供了为活的OCT成像和光遗传起搏准备黑腹果蝇的细节。实验室开发的集成软件处理了成像数据，以创建果蝇心脏功能的视觉呈现和定量特征。结果表明，eNpHR2.0激活引发心脏骤停和心动过缓，ReaChR激活后进行心脏起搏是可行的。
Using Drosophila melanogaster (fruit fly) as a model organism has ensured significant progress in many areas of biological science, from cellular organization and genomic investigations to behavioral studies. Due to the accumulated scientific knowledge, in recent years, Drosophila was brought to the field of modeling human diseases, including heart disorders. The presented work describes the experimental system for monitoring and manipulating the heart function in the context of a whole live organism using red light (617 nm) and without invasive procedures. Control over the heart was achieved using optogenetic tools. Optogenetics combines the expression of light-sensitive transgenic opsins and their optical activation to regulate the biological tissue of interest. In this work, a custom integrated optical coherence tomography (OCT) imaging and optogenetic stimulation system was used to visualize and modulate the functioning D. melanogaster heart at the 3rd instar larval and early pupal developmental stages. The UAS/GAL4 dual genetic system was employed to express halorhodopsin (eNpHR2.0) and red-shifted channelrhodopsin (ReaChR), specifically in the fly heart. Details on preparing D. melanogaster for live OCT imaging and optogenetic pacing are provided. A lab-developed integration software processed the imaging data to create visual presentations and quantitative characteristics of Drosophila heart function. The results demonstrate the feasibility of initiating cardiac arrest and bradycardia caused by eNpHR2.0 activation and performing heart pacing upon ReaChR activation.
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