Multifluorescence labeling techniques and confocal laser scanning microscopy on lung tissue.
Lung tissue consists of more than 40 individual cell types that might interact to produce adverse pathologies. After injury, a number of signaling proteins expressed in various epithelial and other cell types have been linked to the advent of apoptosis, compensatory proliferation, and adaptation to stress. We describe here the use of immunochemistry and multifluorescence approaches using confocal laser scanning microscopy to define the signaling pathways (protein kinases C and mitogen-activated protein kinases) activated by asbestos fibers after inhalation. Using these approaches, we are able to localize signaling events in distinct cell types of the lung and determine their status in the cell cycle (resting or nonresting). Moreover, we are able to determine whether various signaling proteins colocalize in cells and the sites affected by asbestos fibers.
Stern
Maria
M
Department of Pathology, University of Vermont, Burlington, VT, USA.
Taatjes
Douglas J
DJ
Mossman
Brooke T
BT
D000818:Animals; D001194:Asbestos / Q000008:administration & dosage / Q000378:metabolism; D004789:Enzyme Activation; D005456:Fluorescent Dyes / Q000378:metabolism*; D007150:Immunohistochemistry / Q000379:methods; D008168:Lung / Q000166:cytology* / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods*; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D051745:Protein Kinase C-delta / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*
D005456:Fluorescent Dyes; D001194:Asbestos; D051745:Protein Kinase C-delta; D020928:Mitogen-Activated Protein Kinases