Release of cholesterol-rich particles from the macrophage plasma membrane during movement of filopodia and lamellipodia

Abstract

Cultured mouse peritoneal macrophages release large numbers of ~30-nm cholesterol-rich particles. Here, we show that those particles represent fragments of the plasma membrane that are pulled away and left behind during the projection and retraction of filopodia and lamellipodia. Consistent with this finding, the particles are enriched in proteins found in focal adhesions, which attach macrophages to the substrate. The release of particles is abolished by blocking cell movement (either by depolymerizing actin with latrunculin A or by inhibiting myosin II with blebbistatin). Confocal microscopy and NanoSIMS imaging studies revealed that the plasma membrane–derived particles are enriched in 'accessible cholesterol' (a mobile pool of cholesterol detectable with the modified cytolysin ALO-D4) but not in sphingolipid-sequestered cholesterol (a pool detectable with ostreolysin A (OlyA)]. The discovery that macrophages release cholesterol-rich particles during cellular locomotion is likely relevant to cholesterol efflux and could contribute to extracellular cholesterol deposition in atherosclerotic plaques.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files.

Article and author information

Author details

  1. Xuchen Hu

    Department of Medicine, University of California, Los Angeles, Los Angeles, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0944-624X
  2. Thomas A Weston

    Department of Medicine, University of California, Los Angeles, Los Angeles, United States
    Competing interests
    No competing interests declared.
  3. Cuiwen He

    Department of Medicine, University of California, Los Angeles, Los Angeles, United States
    Competing interests
    No competing interests declared.
  4. Rachel S Jung

    Department of Medicine, University of California, Los Angeles, Los Angeles, United States
    Competing interests
    No competing interests declared.
  5. Patrick J Heizer

    Department of Medicine, University of California, Los Angeles, Los Angeles, United States
    Competing interests
    No competing interests declared.
  6. Brian D Young

    Department of Biological Chemistry, University of California, Los Angeles, Los Angeles, United States
    Competing interests
    No competing interests declared.
  7. Yiping Tu

    Department of Medicine, University of California, Los Angeles, Los Angeles, United States
    Competing interests
    No competing interests declared.
  8. Peter Tontonoz

    Department of Pathology and Laboratory Medicine, University of California, Los Angeles, Los Angeles, United States
    Competing interests
    Peter Tontonoz, Reviewing editor, eLife.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1259-0477
  9. James A Wohlschlegel

    Department of Biological Chemistry, University of California, Los Angeles, Los Angeles, United States
    Competing interests
    No competing interests declared.
  10. Haibo Jiang

    Department of Medicine, University of California, Los Angeles, Los Angeles, United States
    For correspondence
    haibo.jiang@uwa.edu.au
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-2384-4826
  11. Stephen G Young

    Department of Medicine, University of California, Los Angeles, Los Angeles, United States
    For correspondence
    sgyoung@mednet.ucla.edu
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-7270-3176
  12. Loren G Fong

    Department of Medicine, University of California, Los Angeles, Los Angeles, United States
    For correspondence
    lfong@mednet.ucla.edu
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-4465-5290

Funding

National Heart, Lung, and Blood Institute (HL090553)

  • Stephen G Young

National Heart, Lung, and Blood Institute (HL087228)

  • Stephen G Young

National Heart, Lung, and Blood Institute (HL125335))

  • Stephen G Young

Fondation Leducq (12CVD04).)

  • Stephen G Young

Ruth L Kirschstein National Research Service Award (T32HL69766))

  • Xuchen Hu

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Fredric B Kraemer, Stanford University, United States

Ethics

Animal experimentation: Animal housing and experimental protocols were approved by UCLA's Animal Research Committee (ARC; 2004-125-51). The animals were housed in an AAALAC (Association for Assessment andAccreditation of Laboratory Animal Care International)-approved facility and cared for accordingto guidelines established by UCLA's Animal Research Committee.

Version history

  1. Received: July 16, 2019
  2. Accepted: September 4, 2019
  3. Accepted Manuscript published: September 5, 2019 (version 1)
  4. Version of Record published: September 18, 2019 (version 2)

Copyright

© 2019, Hu et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Xuchen Hu
  2. Thomas A Weston
  3. Cuiwen He
  4. Rachel S Jung
  5. Patrick J Heizer
  6. Brian D Young
  7. Yiping Tu
  8. Peter Tontonoz
  9. James A Wohlschlegel
  10. Haibo Jiang
  11. Stephen G Young
  12. Loren G Fong
(2019)
Release of cholesterol-rich particles from the macrophage plasma membrane during movement of filopodia and lamellipodia
eLife 8:e50231.
https://doi.org/10.7554/eLife.50231

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https://doi.org/10.7554/eLife.50231