The OptoGenBox - a device for long-term optogenetics in C. elegans.

IF 1.8 4区 医学 Q3 GENETICS & HEREDITY
Journal of neurogenetics Pub Date : 2020-09-01 Epub Date: 2020-06-16 DOI:10.1080/01677063.2020.1776709
Inka Busack, Florian Jordan, Peleg Sapir, Henrik Bringmann
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引用次数: 6

Abstract

Optogenetics controls neural activity and behavior in living organisms through genetically targetable actuators and light. This method has revolutionized biology and medicine as it allows controlling cells with high temporal and spatial precision. Optogenetics is typically applied only at short time scales, for instance to study specific behaviors. Optogenetically manipulating behavior also gives insights into physiology, as behavior controls systemic physiological processes. For example, arousal and sleep affect aging and health span. To study how behavior controls key physiological processes, behavioral manipulations need to occur at extended time scales. However, methods for long-term optogenetics are scarce and typically require expensive compound microscope setups. Optogenetic experiments can be conducted in many species. Small model animals such as the nematode C. elegans have been instrumental in solving the mechanistic basis of medically important biological processes. We developed the OptoGenBox, an affordable stand-alone and simple-to-use device for long-term optogenetic manipulation of C. elegans. The OptoGenBox provides a controlled environment and is programmable to allow the execution of complex optogenetic manipulations over long experimental times of many days to weeks. To test our device, we investigated how optogenetically increased arousal and optogenetic sleep deprivation affect survival of arrested first larval stage C. elegans. We optogenetically activated the nociceptive ASH sensory neurons using ReaChR, thus triggering an escape response and increase in arousal. In addition, we optogenetically inhibited the sleep neuron RIS using ArchT, a condition known to impair sleep. Both optogenetic manipulations reduced survival. Thus, the OptoGenBox presents an affordable system to study the long-term consequences of optogenetic manipulations of key biological processes in C. elegans and perhaps other small animals.

OptoGenBox -秀丽隐杆线虫的长期光遗传学设备。
光遗传学通过基因靶向致动器和光来控制生物体内的神经活动和行为。这种方法可以在时间和空间上精确地控制细胞,给生物学和医学带来了革命性的变化。光遗传学通常只应用于短时间尺度,例如研究特定的行为。光遗传学操纵行为也为生理学提供了见解,因为行为控制着系统的生理过程。例如,觉醒和睡眠影响衰老和健康跨度。为了研究行为如何控制关键的生理过程,行为操纵需要在更长的时间尺度上发生。然而,长期光遗传学的方法是稀缺的,通常需要昂贵的复合显微镜设置。光遗传学实验可以在许多物种中进行。小型模型动物,如线虫秀丽隐杆线虫,在解决医学上重要的生物学过程的机制基础方面发挥了重要作用。我们开发了OptoGenBox,这是一种经济实惠的独立且易于使用的设备,用于秀丽隐杆线虫的长期光遗传学操作。OptoGenBox提供了一个可控的环境,可编程,允许在许多天到几周的长实验时间内执行复杂的光遗传学操作。为了测试我们的装置,我们研究了光遗传学增加唤醒和光遗传学睡眠剥夺如何影响捕获的第一幼虫期秀丽隐杆线虫的存活。我们使用ReaChR光遗传学激活伤害性ASH感觉神经元,从而触发逃避反应和觉醒增加。此外,我们使用ArchT光遗传学抑制睡眠神经元RIS,这是一种已知的损害睡眠的情况。两种光遗传学操作都降低了存活率。因此,OptoGenBox提供了一个经济实惠的系统来研究秀丽隐杆线虫和其他小动物的关键生物过程的光遗传学操作的长期后果。
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来源期刊
Journal of neurogenetics
Journal of neurogenetics 医学-神经科学
CiteScore
4.40
自引率
0.00%
发文量
13
审稿时长
>12 weeks
期刊介绍: The Journal is appropriate for papers on behavioral, biochemical, or cellular aspects of neural function, plasticity, aging or disease. In addition to analyses in the traditional genetic-model organisms, C. elegans, Drosophila, mouse and the zebrafish, the Journal encourages submission of neurogenetic investigations performed in organisms not easily amenable to experimental genetics. Such investigations might, for instance, describe behavioral differences deriving from genetic variation within a species, or report human disease studies that provide exceptional insights into biological mechanisms
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