Analysis of asbestos-induced gene expression changes in bronchiolar epithelial cells using laser capture microdissection and quantitative reverse transcriptase-polymerase chain reaction.
Laser capture microdissection (LCM) enables the removal of discrete microstructures or cell types from properly prepared histological sections. Extraction of RNA from microdissected tissue followed by quantitative reverse transcriptase-polymerase chain (QRT-PCR) reaction permits the analysis of cell-type or microstructure-specific gene expression changes that occur in response to various stimuli in the environment. In our lab, the combination of LCM and QRT-PCR has proven very useful in the determination of the in vivo gene expression changes that occur in bronchiolar epithelium in response to inhalation of crocidolite asbestos. A detailed description of the preparation of cDNA from bronchiolar epithelial cells obtained by LCM is described in this work.
Manning
Christopher B
CB
Department of Pathology, University of Vermont, Burlington, VT, USA.
Mossman
Brooke T
BT
Taatjes
Douglas J
DJ
D017638:Asbestos, Crocidolite / Q000494:pharmacology*; D001980:Bronchi / Q000166:cytology* / Q000502:physiology; D004847:Epithelial Cells / Q000166:cytology / Q000502:physiology*; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D007834:Lasers; D042282:Microdissection / Q000295:instrumentation / Q000379:methods; D008846:Micromanipulation / Q000295:instrumentation / Q000379:methods; D012313:RNA / Q000378:metabolism; D020545:Respiratory Mucosa / Q000166:cytology / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods
D017638:Asbestos, Crocidolite; D012313:RNA