Single Cell Analysis of Phagocytosis, Phagosome Maturation, Phagolysosomal Leakage, and Cell Death Following Exposure of Macrophages to Silica Particles.
Chronic inhalation of silica in various occupational settings results in the development of silicosis, a disease characterized by lung fibrosis. Uptake of silica particles by alveolar macrophages results in cell death and this is one of the contributing factors to the development of silicosis. We have characterized the uncoated or protein-coated (non-opsonized) and Fc receptor-mediated (antibody-opsonized) routes of silica phagocytosis and toxicity. Numerous microscopy techniques and fluorescent probes are outlined in this chapter to carefully measure particle uptake, by macrophages, phagosome maturation, phagosomal reactive oxygen species generation, phagolysosomal leakage, and cell death.
Department of Molecular and Cellular Biology, University of Connecticut, 91 North Eagleville Road, Storrs, CT, 06269, USA.|Department of Pharmaceutical Sciences, University of Connecticut, Storrs, CT, 06269, USA.
Gaurav N
GN
Joshi
Department of Molecular and Cellular Biology, University of Connecticut, 91 North Eagleville Road, Storrs, CT, 06269, USA.|Center for Academic Programs, University of Connecticut, Storrs, CT, 06269, USA.
Renée M
RM
Gilberti
Department of Molecular and Cellular Biology, University of Connecticut, 91 North Eagleville Road, Storrs, CT, 06269, USA. david.knecht@uconn.edu.
David A
DA
Knecht
D000199:Actins / Q000378:metabolism; D000818:Animals; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D005089:Exocytosis / Q000187:drug effects; D005455:Fluorescent Antibody Technique; D049452:Green Fluorescent Proteins / Q000378:metabolism; D021621:Imaging, Three-Dimensional; D008247:Lysosomes / Q000187:drug effects / Q000378:metabolism*; D008264:Macrophages / Q000187:drug effects / Q000473:pathology*; D008561:Membrane Fusion / Q000187:drug effects; D051379:Mice; D009895:Opsonin Proteins / Q000378:metabolism; D010587:Phagocytosis / Q000187:drug effects; D010588:Phagosomes / Q000187:drug effects / Q000378:metabolism*; D010718:Phosphatidylserines / Q000378:metabolism; D011419:Propidium / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D012822:Silicon Dioxide / Q000633:toxicity*; D059010:Single-Cell Analysis / Q000379:methods*; D013194:Staining and Labeling; D013997:Time Factors; D046509:Tissue Culture Techniques
D000199:Actins; D009895:Opsonin Proteins; D010718:Phosphatidylserines; D017382:Reactive Oxygen Species; D049452:Green Fluorescent Proteins; D011419:Propidium; D012822:Silicon Dioxide
Cell death; Endosome; Fluorescence; Lysosome; Microscopy; Phagocytosis; Phagolysosomal leakage; Phagosome; ROS; Silica