Muscle contraction phenotypic analysis enabled by optogenetics reveals functional relationships of sarcomere components in Caenorhabditis elegans.
Muscle contraction phenotypic analysis enabled by optogenetics reveals functional relationships of sarcomere components in Caenorhabditis elegans.
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DOI：
10.1038/srep19900
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发表时间：
2016-01-29
影响因子：
4.6
通讯作者：
Lu H
中科院分区：
文献类型：
--
作者：
Hwang H;Barnes DE;Matsunaga Y;Benian GM;Ono S;Lu H
肌节是肌肉收缩的基本单位，是数百种蛋白质的高度有序复合体。尽管几十年的遗传学研究，这些肌节蛋白在动物行为中的功能关系和作用仍然不清楚。在本文中，我们证明了光遗传学激活的运动神经元，诱导肌肉收缩，可以促进定量研究的肌肉动力学在C。优雅为了增加研究的通量，我们将多个蠕虫并行捕获在微流体装置中，并照射通道视紫红质-2以诱导体壁肌肉收缩。使用图像处理，随着时间的推移量化身体尺寸的变化。从光遗传学测定中提取了总共五个参数，包括收缩和舒张的速率常数，作为肌节功能的描述符。为了潜在地将编码肌节蛋白质的基因在功能上联系起来，在这些参数的基础上进行了层次聚类分析。因为它评估的生理输出不同于传统的测定，这种方法提供了一个补充的表型分析的C。目前在许多实验室中进行的线虫肌肉突变体;这些簇可能提供新的见解，并推动许多肌节组分之间的功能关系的新假设。
The sarcomere, the fundamental unit of muscle contraction, is a highly-ordered complex of hundreds of proteins. Despite decades of genetics work, the functional relationships and the roles of those sarcomeric proteins in animal behaviors remain unclear. In this paper, we demonstrate that optogenetic activation of the motor neurons that induce muscle contraction can facilitate quantitative studies of muscle kinetics in C. elegans. To increase the throughput of the study, we trapped multiple worms in parallel in a microfluidic device and illuminated for photoactivation of channelrhodopsin-2 to induce contractions in body wall muscles. Using image processing, the change in body size was quantified over time. A total of five parameters including rate constants for contraction and relaxation were extracted from the optogenetic assay as descriptors of sarcomere functions. To potentially relate the genes encoding the sarcomeric proteins functionally, a hierarchical clustering analysis was conducted on the basis of those parameters. Because it assesses physiological output different from conventional assays, this method provides a complement to the phenotypic analysis of C. elegans muscle mutants currently performed in many labs; the clusters may provide new insights and drive new hypotheses for functional relationships among the many sarcomere components.