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Originally published as JHC exPRESS on May 6, 2005.
doi:10.1369/jhc.4A6599.2005
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Journal of Histochemistry and Cytochemistry
Volume 53 (8): 1011-1021, 2005
Copyright ©The Histochemical Society, Inc.

Overexpression of eCLCA1 in Small Airways of Horses with Recurrent Airway Obstruction

Friederike Anton, Ina Leverkoehne, Lars Mundhenk, Wallace B. Thoreson and Achim D. Gruber

Department of Pathology, School of Veterinary Medicine Hannover, Hannover, Germany (FA,IL,LM,ADG), and Departments of Ophthalmology & Visual Sciences and Pharmacology, University of Nebraska Medical Center, Durham Research Center, Omaha, Nebraska (WBT)

Correspondence to: Achim D. Gruber, Department of Veterinary Pathology, Free University Berlin, Robert-von-Ostertag-Str. 15, 14163 Berlin, Germany. E-mail: gruber.achim{at}vetmed.fu-berlin.de

The human hCLCA1 and murine mCLCA3 (chloride channels, calcium-activated) have recently been identified as promising therapeutic targets in asthma. Recurrent airway obstruction in horses is an important animal model of human asthma. Here, we have cloned and characterized the first equine CLCA family member, eCLCA1. The 913 amino acids eCLCA1 polypeptide forms a 120-kDa transmembrane glycoprotein that is processed to an 80-kDa protein in vivo. Three single nucleotide polymorphisms were detected in the eCLCA1 coding region in 14 horses, resulting in two amino acid changes (485H/R and 490V/L). However, no functional differences were recorded between the channel properties of the two variants in transfected HEK293 cells. The eCLCA1 protein was detected immunohistochemically in mucin-producing cells in the respiratory and intestinal tracts, cutaneous sweat glands, and renal mucous glands. Strong overexpression of eCLCA1 was observed in the airways of horses with recurrent airway obstruction using Northern blot hybridization, Western blotting, immunohistochemistry, and real-time quantitative RT-PCR. The results suggest that spontaneous or experimental recurrent airway obstruction in horses may serve as a model to study the role of CLCA homologs in chronic airway disease with overproduction of mucins. (J Histochem Cytochem 53:1011–1021, 2005)

Key Words: asthma • chronic obstructive pulmonary • disease • calcium-activated chloride • channels • goblet cells • mucus overproduction


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