An economical and highly adaptable optogenetics system for individual and population-level manipulation of Caenorhabditis elegans.
An economical and highly adaptable optogenetics system for individual and population-level manipulation of Caenorhabditis elegans.
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DOI：
10.1186/s12915-021-01085-2
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发表时间：
2021-08-24
期刊：
影响因子：
5.4
通讯作者：
Nollen EAA
中科院分区：
文献类型：
--
作者：
Koopman M;Janssen L;Nollen EAA
光遗传学允许通过光刺激对可兴奋细胞进行实验操作，而不需要技术上具有挑战性和侵入性的程序。光刺激可以实现高度的空间、时间和强度控制，结合光敏离子通道的细胞类型特异性表达，能够高度特异性和精确地刺激可兴奋细胞。因此，光遗传学工具彻底改变了许多模型中神经元回路的研究，包括秀丽隐杆线虫。尽管存在几个光遗传学系统，允许空间和时间的光敏致动器在C。elegans的高成本和低灵活性限制了光遗传学的广泛应用。在这里，我们开发了一种廉价，易于构建，模块化和可调节的光遗传学设备，用于不同的显微镜和蠕虫追踪器，我们称之为OptoArm。OptoArm允许单蠕虫和多蠕虫照明，并在光强度、照明轮廓和光颜色方面具有自适应性。我们在一项基于人群的多参数研究中展示了OptoArm的力量，该研究涉及运动回路细胞对年龄相关运动性下降的贡献。我们发现，神经肌肉系统的各个组成部分显示出不同的年龄依赖性退化率。胆碱能神经元的功能衰退反映了运动功能的衰退，而GABA能神经元和肌肉细胞相对具有年龄弹性，这表明存在限速细胞并决定神经元回路的老化。我们已经组装了一个经济，可靠，适应性强的光遗传学系统，可以用于解决各种生物学问题。我们提供了一个详细的描述，以及我们的设置的技术和生物学验证的建设。重要的是，OptoArm的使用不仅限于C。elegans，并可能有利于在多个模式生物的研究，使光遗传学更广泛的研究社区。在线版本包含补充材料，可通过10.1186/s12915-021-01085-2获取。
Optogenetics allows the experimental manipulation of excitable cells by a light stimulus without the need for technically challenging and invasive procedures. The high degree of spatial, temporal, and intensity control that can be achieved with a light stimulus, combined with cell type-specific expression of light-sensitive ion channels, enables highly specific and precise stimulation of excitable cells. Optogenetic tools have therefore revolutionized the study of neuronal circuits in a number of models, including Caenorhabditis elegans. Despite the existence of several optogenetic systems that allow spatial and temporal photoactivation of light-sensitive actuators in C. elegans, their high costs and low flexibility have limited wide access to optogenetics. Here, we developed an inexpensive, easy-to-build, modular, and adjustable optogenetics device for use on different microscopes and worm trackers, which we called the OptoArm. The OptoArm allows for single- and multiple-worm illumination and is adaptable in terms of light intensity, lighting profiles, and light color. We demonstrate OptoArm’s power in a population-based multi-parameter study on the contributions of motor circuit cells to age-related motility decline. We found that individual components of the neuromuscular system display different rates of age-dependent deterioration. The functional decline of cholinergic neurons mirrors motor decline, while GABAergic neurons and muscle cells are relatively age-resilient, suggesting that rate-limiting cells exist and determine neuronal circuit ageing. We have assembled an economical, reliable, and highly adaptable optogenetics system which can be deployed to address diverse biological questions. We provide a detailed description of the construction as well as technical and biological validation of our set-up. Importantly, use of the OptoArm is not limited to C. elegans and may benefit studies in multiple model organisms, making optogenetics more accessible to the broader research community. The online version contains supplementary material available at 10.1186/s12915-021-01085-2.
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影响因子：
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作者：
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通讯作者：
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