Heterogeneity of late endosome/lysosomes shown by multiplexed DNA-PAINT imaging.

IF 7.4 1区 生物学 Q1 CELL BIOLOGY
Journal of Cell Biology Pub Date : 2025-01-06 Epub Date: 2024-11-01 DOI:10.1083/jcb.202403116
Charles Bond, Siewert Hugelier, Jiazheng Xing, Elena M Sorokina, Melike Lakadamyali
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引用次数: 0

Abstract

Late endosomes/lysosomes (LELs) are crucial for numerous physiological processes and their dysfunction is linked to many diseases. Proteomic analyses have identified hundreds of LEL proteins; however, whether these proteins are uniformly present on each LEL, or if there are cell-type-dependent LEL subpopulations with unique protein compositions is unclear. We employed quantitative, multiplexed DNA-PAINT super-resolution imaging to examine the distribution of seven key LEL proteins (LAMP1, LAMP2, CD63, Cathepsin D, TMEM192, NPC1, and LAMTOR4). While LAMP1, LAMP2, and Cathepsin D were abundant across LELs, marking a common population, most analyzed proteins were associated with specific LEL subpopulations. Our multiplexed imaging approach identified up to eight different LEL subpopulations based on their unique membrane protein composition. Additionally, our analysis of the spatial relationships between these subpopulations and mitochondria revealed a cell-type-specific tendency for NPC1-positive LELs to be closely positioned to mitochondria. Our approach will be broadly applicable to determining organelle heterogeneity with single organelle resolution in many biological contexts.

多重 DNA-PAINT 成像显示晚期内膜体/溶酶体的异质性。
晚期内体/溶酶体(LELs)对许多生理过程至关重要,其功能障碍与许多疾病有关。蛋白质组学分析已鉴定出数百种 LEL 蛋白;然而,这些蛋白质是否均匀地存在于每个 LEL 上,或者是否存在具有独特蛋白质组成的细胞类型依赖性 LEL 亚群,目前还不清楚。我们采用定量、多重 DNA-PAINT 超分辨率成像技术研究了七种关键 LEL 蛋白(LAMP1、LAMP2、CD63、Cathepsin D、TMEM192、NPC1 和 LAMTOR4)的分布情况。虽然 LAMP1、LAMP2 和 Cathepsin D 在 LEL 中含量丰富,标志着一个共同的群体,但大多数分析的蛋白质都与特定的 LEL 亚群相关。我们的多重成像方法根据 LEL 独特的膜蛋白组成确定了多达八个不同的 LEL 亚群。此外,我们对这些亚群与线粒体之间空间关系的分析表明,NPC1 阳性的 LEL 与线粒体的位置密切相关,这是一种细胞类型特异性趋势。我们的方法将广泛适用于在许多生物环境中以单个细胞器分辨率确定细胞器的异质性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Journal of Cell Biology
Journal of Cell Biology 生物-细胞生物学
CiteScore
12.60
自引率
2.60%
发文量
213
审稿时长
1 months
期刊介绍: The Journal of Cell Biology (JCB) is a comprehensive journal dedicated to publishing original discoveries across all realms of cell biology. We invite papers presenting novel cellular or molecular advancements in various domains of basic cell biology, along with applied cell biology research in diverse systems such as immunology, neurobiology, metabolism, virology, developmental biology, and plant biology. We enthusiastically welcome submissions showcasing significant findings of interest to cell biologists, irrespective of the experimental approach.
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