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JHC exPRESS: First Published May 6, 2005. doi:10.1369/jhc.4A6599.2005
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A more recent version of this article appeared on August 1, 2005.
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ARTICLE

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

Friederike Anton 1, Ina Leverkoehne 1, Lars Mundhenk 1, Wallace B. Thoreson 1 and Achim D. Gruber 1*

1 Department of Pathology, School of Veterinary Medicine 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)

* To whom correspondence should be addressed. E-mail: E-mail: achim.gruber{at}tiho-hannover.de.

Submitted on December 14, 2004
Accepted on 27 March 2005


  Abstract
The human hCLCA1 and murine mCLCA3 (chloride channels, calcium-activated) have recently been identified as promising therapeutic targets in asthma. Recurrent airway obstruction (RAO) 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 (SNPs) were detected in the eCLCA1 coding region in fourteen 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.

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


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