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Phagosome-lysosome fusion. Characterization of intracellular membrane fusion in mouse macrophages

Several approaches have been used to study the determinants of phagosome-lysosome fusion in intact mouse macrophages. Lysosomes were labeled with the fluorescent vital dye acridine orange and the rate and extent of their fusion with yeast-containing phagosomes was monitored by fluorescence microscop...

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Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 1980
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2111457/
https://www.ncbi.nlm.nih.gov/pubmed/7391139
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description Several approaches have been used to study the determinants of phagosome-lysosome fusion in intact mouse macrophages. Lysosomes were labeled with the fluorescent vital dye acridine orange and the rate and extent of their fusion with yeast-containing phagosomes was monitored by fluorescence microscopy. Fusion was also assayed by electron microscopy, using horseradish peroxidase or thorium dioxide as a marker for secondary lysosomes. Good agreemen samples with an enzymatic marker, and thorium dioxide-labeled samples evaluated by stereology. The rate of usion as assayed by fluorescence was not affected by the number of particles ingested, serum concentration, or prior uptake of digestible or nondigestible substances. With this assay it was possible to observe the rate of fusion separate from and uninfluenced by the phagocytic rate. Both the rate and extent of fusion were dramatically increased after several days in culture and similar changes were found by use of the EM assays. Fusion was strongly affected by incubation temperature, having a Q10 of 2.5 No detectable fusion occurred below 15 degrees C, and this inhibition was rapidly reversed when cells were returned to 37 degrees C.
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spelling pubmed-21114572008-05-01 Phagosome-lysosome fusion. Characterization of intracellular membrane fusion in mouse macrophages J Cell Biol Articles Several approaches have been used to study the determinants of phagosome-lysosome fusion in intact mouse macrophages. Lysosomes were labeled with the fluorescent vital dye acridine orange and the rate and extent of their fusion with yeast-containing phagosomes was monitored by fluorescence microscopy. Fusion was also assayed by electron microscopy, using horseradish peroxidase or thorium dioxide as a marker for secondary lysosomes. Good agreemen samples with an enzymatic marker, and thorium dioxide-labeled samples evaluated by stereology. The rate of usion as assayed by fluorescence was not affected by the number of particles ingested, serum concentration, or prior uptake of digestible or nondigestible substances. With this assay it was possible to observe the rate of fusion separate from and uninfluenced by the phagocytic rate. Both the rate and extent of fusion were dramatically increased after several days in culture and similar changes were found by use of the EM assays. Fusion was strongly affected by incubation temperature, having a Q10 of 2.5 No detectable fusion occurred below 15 degrees C, and this inhibition was rapidly reversed when cells were returned to 37 degrees C. The Rockefeller University Press 1980-06-01 /pmc/articles/PMC2111457/ /pubmed/7391139 Text en This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Articles
Phagosome-lysosome fusion. Characterization of intracellular membrane fusion in mouse macrophages
title Phagosome-lysosome fusion. Characterization of intracellular membrane fusion in mouse macrophages
title_full Phagosome-lysosome fusion. Characterization of intracellular membrane fusion in mouse macrophages
title_fullStr Phagosome-lysosome fusion. Characterization of intracellular membrane fusion in mouse macrophages
title_full_unstemmed Phagosome-lysosome fusion. Characterization of intracellular membrane fusion in mouse macrophages
title_short Phagosome-lysosome fusion. Characterization of intracellular membrane fusion in mouse macrophages
title_sort phagosome-lysosome fusion. characterization of intracellular membrane fusion in mouse macrophages
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2111457/
https://www.ncbi.nlm.nih.gov/pubmed/7391139