Characterization of protocadherin-1 expression in primary bronchial epithelial cells: association with epithelial cell differentiation

• Published in: The FASEB journal Vol. 26; no. 1; p. 439
• Main Authors: Koning, Henk, Sayers, Ian, Stewart, Ceri E, de Jong, Debora, Ten Hacken, Nick H T, Postma, Dirkje S, van Oosterhout, Antoon J M, Nawijn, Martijn C, Koppelman, Gerard H
• Format: Journal Article
• Language: English
• Published: United States 01.01.2012
• Subjects: Alternative Splicing - physiology; Bronchi - cytology; Cadherins - chemistry; Cadherins - genetics; Cadherins - metabolism; Cell Differentiation - physiology; Epithelial Cells - cytology; Epithelial Cells - physiology; Gene Expression - physiology; Genetic Variation; Humans; Isomerism; Nucleic Acid Amplification Techniques; Primary Cell Culture; Respiratory Mucosa - cytology; Respiratory Mucosa - physiology; RNA, Messenger - metabolism
• ISSN: 1530-6860
• DOI: 10.1096/fj.11-185207

Protocadherin-1 (PCDH1) is a novel susceptibility gene for asthma that is expressed in airway epithelium. We aimed to characterize PCDH1 mRNA transcripts and protein expression in primary bronchial epithelial cells and to determine regulation of PCDH1 during mucociliary differentiation. Total RNA and protein were isolated from human primary bronchial epithelial cells. PCDH1 transcripts were characterized by rapid amplification of cDNA ends in bronchial epithelial cells of 4 subjects. PCDH1 expression was quantified by quantitative RT-PCR and Western blotting in bronchial epithelial cells directly ex vivo and after air liquid interface (ALI) or submerged culture. We identified 5 novel exons on the 5' end and 1 exon on the 3' end of PCDH1. Novel transcripts showed major variation in expression of intracellular conserved motifs. Expression levels of PCDH1 transcripts encoding exon 1-2 were 4-fold higher, and transcripts encoding exon 3-4 were 15-fold higher in freshly isolated bronchial epithelial cells than in submerged cultures. PCDH1 mRNA (3- to 8-fold) and protein levels (2- to 3-fold) were strongly up-regulated during mucociliary differentiation of primary bronchial epithelial cells in ALI cultures. In summary, PCDH1 transcripts display remarkable variability in expression of conserved intracellular signaling domains. Enhanced PCDH1 expression levels strongly correlate with differentiation of bronchial epithelial cells.