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

Details

• Alternative Title:
  Nat Chem Biol
• Personal Author:
  Gupta, Aarushi ; Rivera-Molina, Felix ; Xi, Zhiqun ; Toomre, Derek ; Schepartz, Alanna
• 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.
• Subjects:
  Biological Transport Carbocyanines Carrier Proteins Cholesterol Endosomes Fibroblasts Fluorescent Dyes HeLa Cells Humans Lysosomes Membrane Glycoproteins Microscopy, Fluorescence Protein Transport

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• Source:
  Nat Chem Biol. 16(4):408-414
• Pubmed ID:
  32094922
• Pubmed Central ID:
  PMC7176048
• Document Type:
  Journal Article
• Funding:
  S10 OD020142/CD/ODCDC CDC HHSUnited States/ ; S10 OD020142/OD/NIH HHSUnited States/ ; R01GM118486/U.S. Department of Health & Human Services ; National Institutes of Health (NIH)International/ ; UL1 TR001863/TR/NCATS NIH HHSUnited States/ ; R35 GM134963/GM/NIGMS NIH HHSUnited States/ ; 5T32GM06754/U.S. Department of Health & Human Services ; National Institutes of Health (NIH)International/ ; R01 GM118486/GM/NIGMS NIH HHSUnited States/ ; P30 DK045735/DK/NIDDK NIH HHSUnited States/ ; R01GM131372/U.S. Department of Health & Human Services ; NIH ; National Institute of General Medical Sciences (NIGMS)International/ ; WT_/Wellcome TrustUnited Kingdom/ ; R01 GM131372/GM/NIGMS NIH HHSUnited States/ ; 095927/A/11/Z/Wellcome Trust (Wellcome)International/
• Volume:
  16
• Issue:
  4
• Collection(s):
  CDC Public Access
• Main Document Checksum:
  urn:sha256:3e906d076e958c0cd3fe98b3d9762f209e1fd0698aeb24aa04385aeb20705226
• Download URL:
  https://stacks.cdc.gov/view/cdc/87158/cdc_87158_DS1.pdf
• File Type:
  [PDF - 1.83 MB ]