Isolation of rodent airway epithelial cell proteins facilitates in vivo proteomics studies of lung toxicity
| Autor: | Lu Zhang, Sharron G. Penn, Alan R. Buckpitt, Charles G. Plopper, Mai-Uyen T. Tran, Dexter Morin, Åsa M. Wheelock |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Male
Proteomics Pulmonary and Respiratory Medicine Pathology medicine.medical_specialty Physiology Immunoblotting Cell Respiratory Mucosa Biology Bronchoalveolar Lavage Rats Sprague-Dawley Metabolomics Cytochrome P-450 Enzyme System In vivo Physiology (medical) medicine Animals Electrophoresis Gel Two-Dimensional Two-dimensional gel electrophoresis Epithelial Cells Cell Biology Actins Epithelium Rats Cell biology Gene expression profiling medicine.anatomical_structure Microscopy Electron Scanning Methacrylates Respiratory epithelium |
| Zdroj: | American Journal of Physiology-Lung Cellular and Molecular Physiology. 286:L399-L410 |
| ISSN: | 1522-1504 1040-0605 |
| DOI: | 10.1152/ajplung.00072.2003 |
| Popis: | 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 that facilitates in vivo proteomics studies of two target-cell pheno-types 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 fivefold higher concentrations and reduces the yield of nonepithelial cell proteins 13-fold compared with homogenates from microdissected airways. In addition, 36% more protein spots were detectable by two-dimensional gel electrophoresis. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|