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Published ahead of print on July 18, 2003, doi:10.1165/rcmb.2003-0134OC
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American Journal of Respiratory Cell and Molecular Biology. Vol. 30, pp. 145-154, 2004
© 2004 American Thoracic Society
DOI: 10.1165/rcmb.2003-0134OC

Expression of Transient Receptor Potential C6 and Related Transient Receptor Potential Family Members in Human Airway Smooth Muscle and Lung Tissue

Randolph L. Corteling, Su Li, June Giddings, John Westwick, Chris Poll and Ian P. Hall

Division of Therapeutics, University Hospital, Queens Medical Centre, Nottingham; and Novartis Respiratory Research Centre, Wimblehurst Road, Horsham, United Kingdom

Address correspondence to: Prof. Ian P. Hall, Division of Therapeutics and Molecular Medicine, South Block, D Floor, University Hospital, Queens Medical Centre, Nottingham NG7 2UH, UK. E-mail: ian.hall{at}nottingham.ac.uk

Elevation of the intracellular free Ca2+ concentration regulates many functional responses in airway smooth muscle, including contraction, proliferation, adhesion, and cell survival. This increase in calcium can be achieved by a release from internal stores (sarcoplasmic reticulum) and/or entry across the cell membrane from the extracellular environment. The molecular identity of this calcium influx pathway in human airway smooth muscle (HASM) remains unclear. Functional studies using Fluo 4-loaded HASM suggest the presence of a histamine H1 receptor-activated Ca2+ entry pathway with characteristics similar to those seen with transient receptor potential (TRP) family homologs. Using a range of molecular and cell biological approaches we defined the expression pattern of transient receptor potential classics (TRPC) homologs in airway cells and tissue. Here we show that HASM and human bronchial epithelial cells both express TRPC1, -4, and -6, with HASM also expressing TRPC3 at the mRNA level. Identification of TRPC6 protein by western blot and confocal microscopy indicated that the protein is localized in specific cell types, suggesting that it plays an important role in regulating key functions in airway cells. These data demonstrate the expression of a range of TRPC homologs in the airway and the presence of a functional Ca2+ entry pathway with characteristics typical of TRPC family members. TRPC homologs may provide an important novel target for the treatment of airway disease.

Abbreviations: airway smooth muscle, ASM • Dulbecco's modified Eagle's medium, DMEM • fetal calf serum, FCS • human airway smooth muscle, HASM • human bronchial epithelial cells, HBEC • phosphate-buffered saline, PBS • polymerase chain reaction, PCR • reverse transcription, RT • Tris-buffered saline, TBS • transient receptor potential, TRP




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