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Published ahead of print on March 30, 2006, doi:10.1165/rcmb.2006-0003OC

Am. J. Respir. Cell Mol. Biol., Volume 35, Number 2, August 2006, 243-251

A more recent version of this article appeared on August 1, 2006
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Submitted on January 3, 2006
Revised on March 28, 2006

Role of TRPC3 in Tumor Necrosis Factor-alpha Enhanced Calcium Influx in Human Airway Myocytes

Thomas A White1, Ailing Xue2, Eduardo N Chini3, Michael Thompson3, Gary C Sieck4, and Mark E Wylam2*

1 Department of Allergy and Immunology, Mayo Clinic College of Medicine, Rochester, MN, USA, 2 Department of Medicine, Mayo Clinic College of Medicine, Rochester, MN, USA, 3 Department of Anesthesiology, Mayo Clinic College of Medicine, Rochester, MN, USA, 4 Department of Physiology and Biomedical Engineering, Mayo Clinic College of Medicine, Rochester, MN, USA

* To whom correspondence should be addressed. E-mail: wylam.mark{at}mayo.edu.

Previous studies have suggested that the pro-inflammatory cytokine, TNF-{alpha}, contributes to airway hyperresponsivness (AHR) by altering airway smooth muscle (ASM) Ca2+ responses to agonist stimulation. The present study examined the effects of TNF-{alpha} on Ca2+ influx pathways in cultured human ASM cells (HASMCs). Proteins encoded by the transient receptor potential (TRP) gene family function as channels through which receptor-operated (ROCE) and store-operated Ca2+ (SOCE) entry occur. In the present study, the presence of TRPC1, TRPC3, TRPC4, TRPC5 and TRPC6 mRNA and protein expression was confirmed in cultured human ASM cells using RT-PCR and Western blot analysis. TNF-{alpha} treatment significantly increased TRPC3 mRNA and protein levels in HASMCs as well as SOCE. TNF-{alpha} treatment also increased both the peak and plateau [Ca2+]i responses in HASMCs elicited by acetylcholine and bradykinin. The effects of TNF-{alpha} treatment on SOCE and agonist-induced [Ca2+]i responses were attenuated using small interfering RNA (siRNA) transfection which knocked down TRPC3 expression. Thus, in inflammatory airway diseases, TNF-{alpha} may result in increased myocyte activation due to altered Ca2+ influx pathways. These results suggest that TRPC3 may be an important therapeutic target in inflammatory airway diseases such as asthma and COPD.




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