Department of Molecular & Cellular Biology, University of California, Berkeley, Berkeley, CA, USA.
Leonard Davis School of Gerontology, University of Southern California, Los Angeles, CA, USA.
Methods Mol Biol. 2022;2364:101-137. doi: 10.1007/978-1-0716-1661-1_5.
The actin cytoskeleton plays a fundamental role in the regulation of multiple cellular pathways, including trafficking and locomotion. The functional integrity of the cytoskeleton is important during aging, as the decline of cytoskeletal integrity contributes to the physiological consequence of aging. Moreover, improving cytoskeletal form and function throughout aging is sufficient to drive life span extension and promote organismal health in multiple model systems. For these reasons, optimized protocols for visualization of the actin cytoskeleton and its downstream consequences on health span and life span are critical for understanding the aging process. In C. elegans, the actin cytoskeleton shows diverse morphologies across tissues, potentially due to the significantly different functions of each cell type. This chapter describes an imaging platform utilizing LifeAct to visualize the actin cytoskeleton in live, whole nematodes throughout the aging process and methods to perform follow-up studies on the life span and health span of these organisms.
肌动蛋白细胞骨架在调节包括运输和运动在内的多种细胞途径中起着基本作用。在衰老过程中，细胞骨架的功能完整性很重要，因为细胞骨架完整性的下降导致衰老的生理后果。此外，在整个衰老过程中改善细胞骨架的形态和功能足以延长寿命并促进多个模型系统中的生物体健康。出于这些原因，优化肌动蛋白细胞骨架及其对健康寿命和寿命影响的可视化方案对于了解衰老过程至关重要。在秀丽隐杆线虫中，肌动蛋白细胞骨架在不同组织中呈现出不同的形态，这可能是由于每种细胞类型的功能显著不同。本章描述了一个利用 LifeAct 在整个衰老过程中对活体线虫进行肌动蛋白细胞骨架成像的成像平台，以及对这些生物体的寿命和健康寿命进行后续研究的方法。
