Endosome motility defects revealed at super-resolution in live cells using HIDE probes.
Aarushi GuptaFelix Rivera-MolinaZhiqun XiDerek K ToomreAlanna SchepartzPublished in: Nature chemical biology (2020)
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.
Keyphrases
- induced apoptosis
- small molecule
- cell cycle arrest
- fluorescence imaging
- living cells
- high density
- single molecule
- endoplasmic reticulum stress
- wild type
- biofilm formation
- cell death
- single cell
- deep learning
- gene expression
- staphylococcus aureus
- smoking cessation
- combination therapy
- visible light
- low density lipoprotein