Endosome motility defects revealed at super-resolution in live cells using HIDE probes.,Nature Chemical Biology

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Endosome motility defects revealed at super-resolution in live cells using HIDE probes.
Nature Chemical Biology ( IF 12.9 ) Pub Date : 2020-02-24 , DOI: 10.1038/s41589-020-0479-z
Aarushi Gupta ₁ , Felix Rivera-Molina ₂ , Zhiqun Xi ₂ , Derek Toomre ₂ , Alanna Schepartz _{1,

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Affiliation

We report new lipid-based, high-density, environmentally sensitive (HIDE) probes that accurately and selectively image endo-lysosomes and their dynamics at super-resolution for extended times. Treatment of live cells with the small molecules DiIC16TCO or DiIC16'TCO followed by in situ tetrazine ligation reaction with the silicon-rhodamine dye SiR-Tz generates the HIDE probes DiIC16-SiR and DiIC16'-SiR in the endo-lysosomal membrane. These new probes support the acquisition of super-resolution videos of organelle dynamics in primary cells for more than 7 min with no detectable change in endosome structure or function. Using DiIC16-SiR and DiIC16'-SiR, we describe direct evidence of endosome motility defects in cells from patients with Niemann-Pick Type-C disease. In wild-type fibroblasts, the probes reveal distinct but rare inter-endosome kiss-and-run events that cannot be observed using confocal methods. Our results shed new light on the role of NPC1 in organelle motility and cholesterol trafficking.

中文翻译：

使用 HIDE 探针在活细胞中以超分辨率显示内体运动缺陷。

我们报告了新的基于脂质的、高密度的、环境敏感的 (HIDE) 探针，该探针可以在更长的时间内以超分辨率准确和选择性地对内溶酶体及其动力学进行成像。用小分子 DiIC16TCO 或 DiIC16'TCO 处理活细胞，然后与硅-罗丹明染料 SiR-Tz 进行原位四嗪连接反应，在内溶酶体膜中生成 HIDE 探针 DiIC16-SiR 和 DiIC16'-SiR。这些新探针支持在原代细胞中获得超过 7 分钟的细胞器动力学超分辨率视频，而内体结构或功能没有可检测到的变化。我们使用 DiIC16-SiR 和 DiIC16'-SiR，描述了 Niemann-Pick C 型疾病患者细胞内体运动缺陷的直接证据。在野生型成纤维细胞中，探针揭示了使用共聚焦方法无法观察到的独特但罕见的内体间亲吻和运行事件。我们的研究结果揭示了 NPC1 在细胞器运动和胆固醇运输中的作用。

更新日期：2020-02-24

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