Analysis of asbestos-induced gene expression changes in bronchiolar epithelial cells using laser capture microdissection and quantitative reverse transcriptase-polymerase chain reaction

Our plain-English summary of Analysis of asbestos-induced gene expression changes in bronchiolar epithelial cells using laser capture microdissection and quantitative reverse transcriptase-polymerase chain reaction, by C.B. Manning, B.T. Mossman, D.J. Taatjes (Methods in Molecular Biology, 2006).

The study

• Authors: C.B. Manning, B.T. Mossman, D.J. Taatjes
• Journal: Methods in Molecular Biology
• Published: 2006;319:231-6
• Cited by: 3 articles, as counted by NIH iCite
• Read it: PubMed · Publisher

Abstract

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.

In plain English

This book chapter was a laboratory methods protocol, not a study reporting new results. The authors described how to collect bronchiolar epithelial cells (cells lining the small airways) from frozen lung sections using laser capture microdissection, then extract RNA from them and convert it to cDNA for measuring gene activity by quantitative reverse transcriptase-polymerase chain reaction. Their stated reason was that the lung has over 40 cell types, so the responses of individual cell types are diluted in whole-lung extracts. The protocol covered preparing animal lungs, cutting 10-micrometer frozen sections, dehydrating them, capturing cells with an infrared laser that melts a polymer film onto the chosen cells, purifying the RNA, removing DNA, and reverse transcription with a "no reverse transcription" control. The species and number of animals were not stated. The authors said the approach had proven very useful in their lab for studying gene expression changes in bronchiolar epithelium after inhalation of crocidolite asbestos. However, this chapter gave no asbestos doses, measurements or findings; that work is cited to a separate publication.

We wrote this plain-English summary from the full article with the help of AI, and a second AI model checked each sentence against the article. It is not the authors' wording; the abstract and the article are their own account.

What the study did and found

• Design: Laboratory methods protocol (book chapter); no new experimental results reported
• Who was studied: Lung tissue from laboratory animals euthanized with sodium pentobarbital; species, number of animals, place and dates not stated
• Exposure: Inhalation of crocidolite asbestos, mentioned as the application of the method in the authors' lab; no doses or exposure details given in this chapter
• The authors' conclusion: The authors stated that combining laser capture microdissection with quantitative RT-PCR had proven very useful in their lab for determining in vivo gene expression changes in bronchiolar epithelium after crocidolite inhalation (no data shown here)

Topics

The National Library of Medicine indexes this study under subjects our site covers: Crocidolite.

Medical terms in this study, explained in our Lexicon: Epithelium, Crocidolite.

Related studies

• Laser capture microdissection reveals dose-response of gene expression in situ consequent to asbestos exposure — Q. Yin, A.R. Brody, et al. (2007)
• Induction of manganese superoxide dismutase gene expression in bronchoepithelial cells after rockwool exposure — J. Marks-Konczalik, A. Gillissen, et al. (1998)
• Changes in DNA 8-hydroxyguanine levels, 8-hydroxyguanine repair activity, and hOGG1 and hMTH1 mRNA expression in human lung alveolar epithelial cells induced by crocidolite asbestos — H.N. Kim, Y. Morimoto, et al. (2001)
• Asbestos induces nuclear factor kappa B (NF-kappa B) DNA-binding activity and NF-kappa B-dependent gene expression in tracheal epithelial cells — Y.M. Janssen, A. Barchowsky, et al. (1995)
• Indications for distinct pathogenic mechanisms of asbestos and silica through gene expression profiling of the response of lung epithelial cells — T.N. Perkins, P.M. Peeters, et al. (2015)
• Crocidolite activates NF-kappa B and MIP-2 gene expression in rat alveolar epithelial cells. Role of mitochondrial-derived oxidants — K.E. Driscoll, J.M. Carter, et al. (1998)

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