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Frustrated Phagocytosis on Micro-Patterned Immune Complexes to Characterize Lysosome Movements in Live Macrophages
Lysosome mobilization is a key cellular process in phagocytes for bactericidal activities and trans-matrix migration. The molecular mechanisms that regulate lysosome mobilization are still poorly known. Lysosomes are hard to track as they move toward phagosomes throughout the cell volume. In order t...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Research Foundation
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3341964/ https://www.ncbi.nlm.nih.gov/pubmed/22566841 http://dx.doi.org/10.3389/fimmu.2011.00051 |
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author | Labrousse, Arnaud M. Meunier, Etienne Record, Julien Labernadie, Anna Beduer, Amélie Vieu, Christophe Ben Safta, Thouraya Maridonneau-Parini, Isabelle |
author_facet | Labrousse, Arnaud M. Meunier, Etienne Record, Julien Labernadie, Anna Beduer, Amélie Vieu, Christophe Ben Safta, Thouraya Maridonneau-Parini, Isabelle |
author_sort | Labrousse, Arnaud M. |
collection | PubMed |
description | Lysosome mobilization is a key cellular process in phagocytes for bactericidal activities and trans-matrix migration. The molecular mechanisms that regulate lysosome mobilization are still poorly known. Lysosomes are hard to track as they move toward phagosomes throughout the cell volume. In order to anticipate cell regions where lysosomes are recruited to, human and RAW264.7 macrophages were seeded on surfaces that were micro-patterned with immune complexes (ICs) as 4 μm-side squares. Distances between IC patterns were adapted to optimize cell spreading in order to constrain lysosome movements mostly in two dimensions. FcΓ receptors triggered local frustrated phagocytosis, frustrated phagosomes appeared as rings of F-actin dots around the IC patterns as early as 5 min after cells made contact with the substratum. Frustrated phagosomes recruited actin-associated proteins (vinculin, paxillin, and gelsolin). The fusion of lysosomes with frustrated phagosomes was shown by the release of beta-hexosaminidase and the recruitment of Lamp1 to frustrated phagosomes. Lysosomes of RAW264.7 macrophages were labeled with cathepsin-D-mCherry to visualize their movements toward frustrated phagosomes. Lysosomes saltatory movements were markedly slowed down compared to cells layered on non-opsonized patterns. In addition, the linearity of the trajectories and the frequency and duration of contacts of lysosomes with frustrated phagosomes were measured. Our experimental set-up is the first step toward deciphering molecular mechanisms which are involved in lysosome movements in the cytoplasm (speed, directionality, and interaction with phagosomes), and opens the door to approaches such as RNA interference, pharmacological inhibition, or mutant expression. |
format | Online Article Text |
id | pubmed-3341964 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Frontiers Research Foundation |
record_format | MEDLINE/PubMed |
spelling | pubmed-33419642012-05-07 Frustrated Phagocytosis on Micro-Patterned Immune Complexes to Characterize Lysosome Movements in Live Macrophages Labrousse, Arnaud M. Meunier, Etienne Record, Julien Labernadie, Anna Beduer, Amélie Vieu, Christophe Ben Safta, Thouraya Maridonneau-Parini, Isabelle Front Immunol Immunology Lysosome mobilization is a key cellular process in phagocytes for bactericidal activities and trans-matrix migration. The molecular mechanisms that regulate lysosome mobilization are still poorly known. Lysosomes are hard to track as they move toward phagosomes throughout the cell volume. In order to anticipate cell regions where lysosomes are recruited to, human and RAW264.7 macrophages were seeded on surfaces that were micro-patterned with immune complexes (ICs) as 4 μm-side squares. Distances between IC patterns were adapted to optimize cell spreading in order to constrain lysosome movements mostly in two dimensions. FcΓ receptors triggered local frustrated phagocytosis, frustrated phagosomes appeared as rings of F-actin dots around the IC patterns as early as 5 min after cells made contact with the substratum. Frustrated phagosomes recruited actin-associated proteins (vinculin, paxillin, and gelsolin). The fusion of lysosomes with frustrated phagosomes was shown by the release of beta-hexosaminidase and the recruitment of Lamp1 to frustrated phagosomes. Lysosomes of RAW264.7 macrophages were labeled with cathepsin-D-mCherry to visualize their movements toward frustrated phagosomes. Lysosomes saltatory movements were markedly slowed down compared to cells layered on non-opsonized patterns. In addition, the linearity of the trajectories and the frequency and duration of contacts of lysosomes with frustrated phagosomes were measured. Our experimental set-up is the first step toward deciphering molecular mechanisms which are involved in lysosome movements in the cytoplasm (speed, directionality, and interaction with phagosomes), and opens the door to approaches such as RNA interference, pharmacological inhibition, or mutant expression. Frontiers Research Foundation 2011-10-12 /pmc/articles/PMC3341964/ /pubmed/22566841 http://dx.doi.org/10.3389/fimmu.2011.00051 Text en Copyright © 2011 Labrousse, Meunier, Record, Labernadie, Beduer, Vieu, Ben Safta and Maridonneau-Parini. http://www.frontiersin.org/licenseagreement This is an open-access article subject to a non-exclusive license between the authors and Frontiers Media SA, which permits use, distribution and reproduction in other forums, provided the original authors and source are credited and other Frontiers conditions are complied with. |
spellingShingle | Immunology Labrousse, Arnaud M. Meunier, Etienne Record, Julien Labernadie, Anna Beduer, Amélie Vieu, Christophe Ben Safta, Thouraya Maridonneau-Parini, Isabelle Frustrated Phagocytosis on Micro-Patterned Immune Complexes to Characterize Lysosome Movements in Live Macrophages |
title | Frustrated Phagocytosis on Micro-Patterned Immune Complexes to Characterize Lysosome Movements in Live Macrophages |
title_full | Frustrated Phagocytosis on Micro-Patterned Immune Complexes to Characterize Lysosome Movements in Live Macrophages |
title_fullStr | Frustrated Phagocytosis on Micro-Patterned Immune Complexes to Characterize Lysosome Movements in Live Macrophages |
title_full_unstemmed | Frustrated Phagocytosis on Micro-Patterned Immune Complexes to Characterize Lysosome Movements in Live Macrophages |
title_short | Frustrated Phagocytosis on Micro-Patterned Immune Complexes to Characterize Lysosome Movements in Live Macrophages |
title_sort | frustrated phagocytosis on micro-patterned immune complexes to characterize lysosome movements in live macrophages |
topic | Immunology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3341964/ https://www.ncbi.nlm.nih.gov/pubmed/22566841 http://dx.doi.org/10.3389/fimmu.2011.00051 |
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