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Endosome motility defects revealed at super-resolution in live cells using HIDE probes.


ABSTRACT: 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.

SUBMITTER: Gupta A 

PROVIDER: S-EPMC7176048 | biostudies-literature | 2020 Apr

REPOSITORIES: biostudies-literature

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Endosome motility defects revealed at super-resolution in live cells using HIDE probes.

Gupta Aarushi A   Rivera-Molina Felix F   Xi Zhiqun Z   Toomre Derek D   Schepartz Alanna A  

Nature chemical biology 20200224 4


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<sub>16</sub>TCO or DiIC<sub>16'</sub>TCO followed by in situ tetrazine ligation reaction with the silicon-rhodamine dye SiR-Tz generates the HIDE probes DiIC<sub>16</sub>-SiR and DiIC<sub>16'</sub>-SiR in the endo-lysosomal membrane. These new probes su  ...[more]

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