Kim Juri, Dutta Naibedya, Vega Matthew, Bong Andrew, Averbukh Maxim, Aviles Barahona Rebecca, Alcala Athena, Holmes Jacob T, Garcia Gilberto, Higuchi-Sanabria Ryo
Leonard Davis School of Gerontology, University of Southern California, Los Angeles, CA, USA.
Leonard Davis School of Gerontology, University of Southern California, Los Angeles, CA, USA
Life Sci Alliance. 2025 Jun 3;8(8). doi: 10.26508/lsa.202403189. Print 2025 Aug.
Measurements of mitochondrial morphology are a powerful proxy for assessing mitochondrial health, particularly during aging when organelle dynamics are disrupted. provides an ideal system for in vivo mitochondrial imaging, but widely used high-copy transgenic strains can induce artifacts that confound interpretation because of their impact on cellular and organismal health and physiology. Here, we present and validate a suite of strains expressing single-copy, matrix-localized GFP in the muscle, intestine, and hypodermis using the MosSCI technology. These strains enable robust, tissue-specific visualization of mitochondrial morphology without the caveats associated with multi-copy reporters. We benchmark their performance against existing models and demonstrate that our mitochondrial reporters are similarly capable of assessing age-associated mitochondrial morphology, while avoiding defects in cellular and physiological health associated with the multi-copy reporters. Furthermore, we assess how aging methods, bacterial diets, and inhibition of fusion and fission machinery impact mitochondrial morphology during aging. Our findings provide a standardized and physiologically relevant platform for studying mitochondrial dynamics during aging in .
线粒体形态测量是评估线粒体健康状况的有力指标，尤其是在衰老过程中细胞器动态被破坏时。秀丽隐杆线虫为体内线粒体成像提供了理想的系统，但广泛使用的高拷贝转基因品系会因对细胞和机体健康及生理的影响而诱导混淆解释的假象。在此，我们展示并验证了一组利用MosSCI技术在肌肉、肠道和皮下表达单拷贝、定位于线粒体基质的绿色荧光蛋白（GFP）的品系。这些品系能够实现线粒体形态的稳健、组织特异性可视化，而不存在与多拷贝报告基因相关的问题。我们将它们的性能与现有模型进行了对比，证明我们的线粒体报告基因同样能够评估与年龄相关的线粒体形态，同时避免了与多拷贝报告基因相关的细胞和生理健康缺陷。此外，我们评估了衰老方法、细菌饮食以及融合和裂变机制的抑制如何在衰老过程中影响线粒体形态。我们的研究结果为研究秀丽隐杆线虫衰老过程中的线粒体动态提供了一个标准化且与生理相关的平台。
