Published ahead of print on August 28, 2003, doi:10.1165/rcmb.2003-0214OC Am. J. Respir. Cell Mol. Biol., Volume 30, Number 3, March 2004, 296-310 A more recent version of this article appeared on March 1, 2004
Submitted on June 10, 2003 Gene Expression Analysis in Response to Lung Toxicants: I. Sequencing and Microarray DevelopmentMichael A Shultz1*,1 Global Research, Amersham Biosciences (SV) Corp., Sunnyvale, CA, USA; Veterinary Medicine: Molecular Biosciences, University of California, Davis, Davis, CA, USA, 2 Global Research, Amersham Biosciences (SV) Corp., Sunnyvale, CA, USA, 3 Veterinary Medicine: Anatomy, Physiology, and Cell Biology, University of California, Davis, Davis, CA, USA, 4 Veterinary Medicine: Molecular Biosciences, University of California, Davis, Davis, CA, USA * To whom correspondence should be addressed. E-mail: mashultz{at}ucdavis.edu.
A key challenge in measuring gene expression changes in the lung in response to site-selective toxicants is differentiating between target and nontarget areas. The toxicity for the cytotoxicant 1-nitronaphthalene is highly localized in the airway epithelium. Target cells comprise but a fraction of the total lung cell mass; measurements from whole lung homogenates are not likely to reflect what occurs at the target site. Additionally, the use of generic microarrays to measure expression in airway epithelium may not provide a good representation of transcripts present at the site of toxic action. cDNA libraries from airway and alveolar subcompartments of rat lung were sequenced for the development of a custom microarray representative of these lung regions. We identified 7,460 non-redundant rat lung sequences. Nearly 30% of the sequences on this array are not present on the Affymetrix Rat GeneChip 230. A 20,000-element microarray was developed which delineates differences in gene expression between subcompartments. This is the first in a series of papers employing this microarray for detecting gene expression changes during acute injury produced by 1-nitronaphthalene and subsequent repair.
|
|||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
