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Am J Physiol Lung Cell Mol Physiol (October 31, 2003). doi:10.1152/ajplung.00072.2003
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Submitted on March 13, 2003
Accepted on October 27, 2003

Isolation of rodent airway epithelial cell proteins facilitates in vivo proteomics studies of lung toxicity

Asa M. Wheelock1*, Lu Zhang2, Mai-Uyen Tran3, Dexter Morin1, Sharron Penn2, Alan R. Buckpitt1, and Charles G. Plopper3

1 Department of Molecular Biosciences, School of Veterinary Medicine, University of California, Davis, Davis, CA, USA
2 Advanced Research Team, Amersham Biosciences Corp., Sunnyvale, CA, USA
3 Department of Anatomy, Physiology and Cell Biology, School of Veterinary Medicine, University of California, Davis, Davis, CA, USA

* To whom correspondence should be addressed. E-mail: amkarlsson{at}ucdavis.edu.

Recent developments in genomics, proteomics, and metabolomics hold substantial promise for understanding cellular responses to toxicants. Gene expression profiling is now considered standard procedure, but numerous publications reporting a lack of correlation between mRNA and protein expression emphasize the importance of conducting parallel proteomics studies. The cellular complexity of the lung presents great challenges for in vivo proteomics and improved isolation methods for proteins from specific lung cell-phenotypes are required. To address this issue, we have developed a novel method for isolation of rodent airway epithelial cell proteins, which facilitates in vivo proteomics studies of two target-cell phenotypes of the lung, Clara cells and ciliated cells. The airway epithelial cell proteins are reproducibly solubilized, leaving the underlying basement membrane and smooth muscle intact as shown by histopathological analyses. The method yields epithelial cell-specific proteins in 5-fold higher concentrations and reduces the yield of non-epithelial cell proteins 13-fold in comparison to homogenates from microdissected airways. In addition, 36% more protein spots were detectable by 2-dimensional gel electrophoresis.




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