Investigating the correlation of muscle function tests and sarcomere organization in C. elegans.
Investigating the correlation of muscle function tests and sarcomere organization in C. elegans.
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DOI：
10.1186/s13395-021-00275-4
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发表时间：
2021-08-13
期刊：
影响因子：
4.9
通讯作者：
Vanapalli SA
中科院分区：
文献类型：
--
作者：
Lesanpezeshki L;Qadota H;Darabad MN;Kashyap K;Lacerda CMR;Szewczyk NJ;Benian GM;Vanapalli SA
秀丽隐杆线虫已被广泛用作研究肌肉结构和功能的模型。其体壁肌肉在功能和结构上与脊椎动物骨骼肌相似，具有保守的分子通路，有助于肌节结构和肌肉功能。然而，对肌力和肌节组织之间的关系缺乏系统的研究。本文通过研究各种肌节蛋白和膜附着组分对肌肉结构和功能的贡献，引入秀丽隐杆线虫作为模型生物，研究肌肉力量的遗传基础。我们采用两种最近发展的分析，包括肌肉力量的发挥，以研究肌肉功能与肌节组织的相关性。我们使用了一种名为NemaFlex的微流控柱平台，该平台可以量化最大可承受力，并进行挖洞试验，挑战动物在化学刺激下进行三维移动。我们选择了20个在肌节的不同亚结构中存在已知缺陷的突变体，并比较了产生和传递力所需的肌肉蛋白的生理功能。我们还使用免疫染色方法表征了肌节的破坏程度。我们发现，在细丝、粗丝和m系中具有遗传缺陷的突变体通常较弱，我们的检测方法成功地检测了每个肌节检测位置的功能变化。我们发现NemaFlex和挖洞试验在功能上是不同的，可以对肌肉生理学的不同方面提供信息。具体来说，挖洞法在肌肉突变表型分析中具有更大的带宽，因为它可以在NemaFlex中施加正常肌肉力的同时选择10个额外的受损突变体。这使我们能够结合他们的读数来开发一个综合肌肉功能评分，该评分被发现与肌肉结构紊乱的评分相关。我们的研究结果强调了NemaFlex和穴居试验对秀丽隐杆线虫肌肉生理学评价的适用性。利用这些方法，我们讨论了所研究的肌节蛋白对肌肉功能和结构的重要性。我们开发的评分方法增强了秀丽隐杆线虫作为研究肌肉功能的遗传模型的效用。在线版本包含补充材料，可在10.1186/s13395-021-00275-4获得。
Caenorhabditis elegans has been widely used as a model to study muscle structure and function. Its body wall muscle is functionally and structurally similar to vertebrate skeletal muscle with conserved molecular pathways contributing to sarcomere structure, and muscle function. However, a systematic investigation of the relationship between muscle force and sarcomere organization is lacking. Here, we investigate the contribution of various sarcomere proteins and membrane attachment components to muscle structure and function to introduce C. elegans as a model organism to study the genetic basis of muscle strength. We employ two recently developed assays that involve exertion of muscle forces to investigate the correlation of muscle function to sarcomere organization. We utilized a microfluidic pillar-based platform called NemaFlex that quantifies the maximum exertable force and a burrowing assay that challenges the animals to move in three dimensions under a chemical stimulus. We selected 20 mutants with known defects in various substructures of sarcomeres and compared the physiological function of muscle proteins required for force generation and transmission. We also characterized the degree of sarcomere disorganization using immunostaining approaches. We find that mutants with genetic defects in thin filaments, thick filaments, and M-lines are generally weaker, and our assays are successful in detecting the functional changes in response to each sarcomere location tested. We find that the NemaFlex and burrowing assays are functionally distinct informing on different aspects of muscle physiology. Specifically, the burrowing assay has a larger bandwidth in phenotyping muscle mutants, because it could pick ten additional mutants impaired while exerting normal muscle force in NemaFlex. This enabled us to combine their readouts to develop an integrated muscle function score that was found to correlate with the score for muscle structure disorganization. Our results highlight the suitability of NemaFlex and burrowing assays for evaluating muscle physiology of C. elegans. Using these approaches, we discuss the importance of the studied sarcomere proteins for muscle function and structure. The scoring methodology we have developed enhances the utility of C. elegans as a genetic model to study muscle function. The online version contains supplementary material available at 10.1186/s13395-021-00275-4.
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DOI：
10.1083/jcb.132.5.835
发表时间：
1996-03
期刊：
The Journal of cell biology
影响因子：
--
作者：
Benian GM;Tinley TL;Tang X;Borodovsky M
通讯作者：
Borodovsky M
DOI：
10.1083/jcb.101.4.1532
发表时间：
1985-10
期刊：
The Journal of cell biology
影响因子：
--
作者：
Francis GR;Waterston RH
通讯作者：
Waterston RH
影响因子：
3.3
作者：
Han, Hsiao-Fen;Beckerle, Mary C.
通讯作者：
Beckerle, Mary C.
影响因子：
5.2
作者：
Gaffney, Christopher J.;Pollard, Amelia;Szewczyk, Nathaniel J.
通讯作者：
Szewczyk, Nathaniel J.
影响因子：
64.8
作者：
Herndon, LA;Schmeissner, PJ;Driscoll, M
通讯作者：
Driscoll, M