Site-specific Differences in Gene Expression of Secreted Proteins in the Mouse Lung: Comparison of Methods to Show Differences by Location

• Published in: The journal of histochemistry and cytochemistry 2010-12, Vol.58 (12), p.1107-1119
• Main Authors: Sutherland, Katherine M., Combs, Trenton J., Edwards, Patricia C., Laura, S. Van Winkle
• Format: Article
• Language: English
• Subjects: Animals; Calcitonin Gene-Related Peptide - biosynthesis; Calcitonin Gene-Related Peptide - genetics; Calcitonin Gene-Related Peptide - secretion; Gene Expression Profiling; Immunohistochemistry; Lung - metabolism; Male; Mice; Organ Specificity; Proteins - genetics; Proteins - metabolism; Proteins - secretion; Pulmonary Surfactant-Associated Protein A - biosynthesis; Pulmonary Surfactant-Associated Protein A - genetics; Pulmonary Surfactant-Associated Protein A - secretion; Pulmonary Surfactant-Associated Protein C - biosynthesis; Pulmonary Surfactant-Associated Protein C - genetics; Pulmonary Surfactant-Associated Protein C - secretion; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - biosynthesis; RNA, Messenger - genetics; Uteroglobin - biosynthesis; Uteroglobin - genetics; Uteroglobin - secretion
• ISSN: 0022-1554; 1551-5044
• DOI: 10.1369/jhc.2010.956052

Studies on the effects of pulmonary toxicants on the lung often overlook the fact that site-specific changes are likely to occur in response to chemical exposure. These changes can be highly focal and may be undetected by methods that do not examine specific lung regions. This problem is especially acute for studies of the conducting airways. In this study, differential gene expression of secreted proteins in the lung by different methods of collection (whole lung, gross airway microdissection, and laser capture microdissection) and by airway levels (whole lobe, whole airway tree, proximal airways, airway bifurcations, and terminal bronchioles) was examined. Site-specific sampling approaches were combined with methods to detect both gene and corresponding protein expression in different lung regions. Differential expression of mRNA by both airway level and lung region was determined for Clara cell secretory protein, calcitonin gene-related peptide, uteroglobin-related protein 2, surfactant protein A, and surfactant protein C. Therefore, for maximal enrichment of mRNA and maximal ability to identify changes in mRNA levels in the diseased state or in response to chemical exposure, it is critical to choose the appropriate airway region and sample collection method to enrich detection of the transcript(s) of interest. (J Histochem Cytochem 58:1107–1119, 2010)