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Endosome motility defects revealed at super-resolution in live cells using HIDE probes
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4 2020
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Source: Nat Chem Biol. 16(4):408-414
[PDF-1.83 MB]
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Alternative Title:Nat Chem Biol
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Description: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|TCO or DiIC|TCO followed by in situ tetrazine ligation reaction with the silicon-rhodamine dye SiR-Tz generates the HIDE probes DiIC|-SiR and DiIC|-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|-SiR and DiIC|-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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Pubmed ID:32094922
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Pubmed Central ID:PMC7176048
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