Kinfe TM (2000)
Publication Type: Journal article
Publication year: 2000
Book Volume: 68
Journal Issue: 4-5
DOI: 10.1159/000055922
Objective: Cell type-specific mRNA quantitation can be reliably performed after harvesting less than 20 cell profiles from haemalaun-stained cryosections by laser-assisted cell picking. Up to now it has been unclear to what extent these techniques can be used to analyze differential gene expression in complex tissues. Methods: Using a rat model of experimental endotoxin priming of the lung various pulmonary cell types were microdissected from isolated perfused and ventilated rat lungs after aerosol lipopolysaccharide/interferon-γ stimulation. Results: Porphobilinogen deaminase housekeeping gene (PBGD) and nitric oxide synthase II (NOSII) mRNA in arterial endothelial cells (AEC), bronchiolar epithelial cells (BEC), alveolar septum containing monocytes/macrophages (AS+), alveolar septum without monocytes/macrophages (AS–) and intraluminar alveolar macrophages (AM) could be quantified by real-time RT-PCR. The strongest upregulation of NOSII mRNA occurred in AM, while minimal NOSII expression was detected in BEC, AS+ and AS–. In AEC NOSII mRNA was not detectable. Conclusion: The combination of laser microdissection and real-time RT-PCR is a valuable tool for the quantitative in situ characterization of differential gene expression within complex tissues.
APA:
Kinfe, T.M. (2000). Cell type-specific mRNA quantitation in non-neoplastic tissues after laser-assisted cell picking. Pathobiology, 68(4-5). https://doi.org/10.1159/000055922
MLA:
Kinfe, Thomas Mehari. "Cell type-specific mRNA quantitation in non-neoplastic tissues after laser-assisted cell picking." Pathobiology 68.4-5 (2000).
BibTeX: Download