Cytoskeletal reorganizations in human umbilical vein endothelial cells as a result of cytokine exposure.
Treatment of HUVECs in culture with several cytokines and phorbol esters caused reorganizations of the actin and microtubule networks, as well as a redistribution of focal contract proteins. However, expression of the cytoskeletal proteins which link cells, via integrins, to the substrate, was not significantly affected. Indirect immunofluorescence microscopy of endothelial cells after treatment with interleukin-1 alpha and beta, gamma-interferon, tumor necrosis factor (TNF), phorbol 12-myristate 13-acetate, and phorbol 12,13-dibutyrate allowed us to observe reductions in the areas of cell-cell contact, redistribution of the stress fiber network, and concomitant changes in focal contacts. Microtubule arrays in TNF-treated cells became bundled. Phorbol esters induced formation of microtubule organizing centers not seen in resting or TNF-treated HUVECs. Talin was distributed along stress fibers and not exclusively in focal contacts. Vitronectin receptor was observed in focal contacts, occasionally at cell-cell contacts, and in vesicular structures close to the lumenal surface, after both types of treatment. Although these morphological changes were easily observed by indirect immunofluorescence, no quantitative differences in specific cytoskeletal proteins were detected by immunoblots and [35S]cysteine metabolic labeling experiments.
Molony
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Department of Cell Biology, Glaxo Research Institute, Research Triangle Park, North Carolina 27709.
Armstrong
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D015815:Cell Adhesion Molecules / Q000378:metabolism; D002450:Cell Communication / Q000187:drug effects; D003545:Cysteine / Q000378:metabolism; D016207:Cytokines / Q000494:pharmacology*; D003598:Cytoskeletal Proteins / Q000378:metabolism; D003599:Cytoskeleton / Q000187:drug effects* / Q000378:metabolism / Q000648:ultrastructure; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D006801:Humans; D015151:Immunoblotting; D007371:Interferon-gamma / Q000494:pharmacology; D007375:Interleukin-1 / Q000494:pharmacology; D008870:Microtubules / Q000187:drug effects / Q000648:ultrastructure; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology; D011971:Receptors, Immunologic / Q000648:ultrastructure; D019030:Receptors, Vitronectin; D013462:Sulfur Radioisotopes; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D015815:Cell Adhesion Molecules; D016207:Cytokines; D003598:Cytoskeletal Proteins; D007375:Interleukin-1; D011971:Receptors, Immunologic; D019030:Receptors, Vitronectin; D013462:Sulfur Radioisotopes; D014409:Tumor Necrosis Factor-alpha; D015240:Phorbol 12,13-Dibutyrate; D007371:Interferon-gamma; D003545:Cysteine; D013755:Tetradecanoylphorbol Acetate