Sarasija Shaarika, Norman Kenneth R
Department of Regenerative and Cancer Cell Biology, Albany Medical College.
Department of Regenerative and Cancer Cell Biology, Albany Medical College;
J Vis Exp. 2019 Feb 23(144). doi: 10.3791/59277.
Optimal mitochondrial function is critical for healthy cellular activity, particularly in cells that have high energy demands like those in the nervous system and muscle. Consistent with this, mitochondrial dysfunction has been associated with a myriad of neurodegenerative diseases and aging in general. Caenorhabditis elegans have been a powerful model system for elucidating the many intricacies of mitochondrial function. Mitochondrial respiration is a strong indicator of mitochondrial function and recently developed respirometers offer a state-of-the-art platform to measure respiration in cells. In this protocol, we provide a technique to analyze live, intact C. elegans. This protocol spans a period of ~7 days and includes steps for (1) growing and synchronization of C. elegans, (2) preparation of compounds to be injected and hydration of probes, (3) drug loading and cartridge equilibration, (4) preparation of worm assay plate and assay run, and (5) post-experiment data analysis.
最佳线粒体功能对于健康的细胞活动至关重要,尤其是在像神经系统和肌肉中那些有高能量需求的细胞里。与此一致的是,线粒体功能障碍总体上与无数神经退行性疾病和衰老相关。秀丽隐杆线虫一直是阐明线粒体功能诸多复杂细节的强大模型系统。线粒体呼吸是线粒体功能的一个有力指标,最近开发的呼吸计提供了一个用于测量细胞呼吸的先进平台。在本方案中,我们提供一种分析活的、完整的秀丽隐杆线虫的技术。本方案跨度约7天,包括以下步骤:(1)秀丽隐杆线虫的培养与同步化,(2)待注射化合物的制备及探针的水化,(3)药物加载和小室平衡,(4)线虫检测板的制备及检测运行,以及(5)实验后数据分析。