期刊
NATURE CHEMICAL BIOLOGY
卷 16, 期 4, 页码 408-+出版社
NATURE PORTFOLIO
DOI: 10.1038/s41589-020-0479-z
关键词
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资金
- NIH [5T32GM06754 3-12, R01GM131372-01, R01GM118486, S10 OD020142]
- Wellcome Trust [095927/A/11/Z]
- Wellcome Trust [095927/A/11/Z] Funding Source: Wellcome Trust
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 DiIC(16)TCO or DiIC(16')TCO followed by in situ tetrazine ligation reaction with the silicon-rhodamine dye SiR-Tz generates the HIDE probes DiIC(16)-SiR and DiIC(16')-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 DiIC(16)-SiR and DiIC(16')-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.
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