The OptoGenBox - a device for long-term optogenetics inC. elegans
The OptoGenBox - a device for long-term optogenetics inC. elegans
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DOI：
10.1080/01677063.2020.1776709
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发表时间：
2020-06-13
影响因子：
1.9
通讯作者：
Bringmann, Henrik
中科院分区：
文献类型：
--
作者：
Busack, Inka;Jordan, Florian;Bringmann, Henrik
光遗传学通过遗传靶向致动器和光控制生物体的神经活动和行为。这种方法彻底改变了生物学和医学，因为它允许以高时间和空间精度控制细胞。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给生理学带来了深刻的见解，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寿命。为了研究行为如何控制关键的生理过程，行为操纵需要在延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。小型模式动物如线虫C. elegans已经在解决医学上重要的生物过程的机械基础方面发挥了作用。我们开发了OptoGenBox，这是一种负担得起的独立和简单易用的设备，用于长期的光遗传学操作。优雅OptoGenBox提供了一个受控的环境，并且是可编程的，以允许在许多天到数周的长实验时间内执行复杂的光遗传学操作。为了测试我们的设备，我们研究了光遗传学增加的唤醒和光遗传学睡眠剥夺如何影响被捕的第一幼虫stagecc的存活。优雅我们使用ReaChR光遗传学激活伤害性ASH感觉神经元，从而触发逃避反应并增加唤醒。此外，我们使用ArchT光遗传学抑制睡眠神经元RIS，这是一种已知会损害睡眠的条件。两种光遗传学操作都降低了存活率。因此，OptoGenBox提供了一个负担得起的系统来研究C中关键生物过程的光遗传学操纵的长期后果。elegans也许and perhaps其他little animals动物.
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 nematodeC. eleganshave 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 ofC. 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 stageC. 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 inC. elegansand perhaps other small animals.