Evaluation of MUC5AC expression and upregulation in airway epithelial cells of horses

Karen L. Oslund, Grete Adamson, Reen Wu

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Objective - To isolate and culture primary equine airway epithelial cells in vitro and elucidate the major cytokines involved in expression of the gel-forming mucin gene MUC5AC in horses. Sample Population - 12 tracheas obtained within 5 hours after euthanasia from horses free from respiratory tract disease. Procedures - Tracheal rings were digested overnight in 0.2% protease, and dissociated airway epithelial cells were grown in a serum-free defined medium at an air-liquid interface until confluence was achieved. Differentiated airway epithelial cells were treated with a panel of recombinant equine cytokines followed by quantitative reverse transcriptase PCR assay for mRNA of equine MUC5AC and the control gene glyceraldehyde 3-phosphate dehydrogenase. Cultures were incubated in the presence of isohelenin, a nuclear factor κB-DNA-binding inhibitor, to investigate transcriptional regulation of MUC5AC. Results - Light and electron microscopy revealed a differentiated epithelium with ciliated cells, nonciliated mucous cells, and basal-like cells. Recombinant equine tumor necrosis factor-α was the major mediator in the cytokine panel that significantly increased MUC5AC mRNA by a factor of 5 in a dose- and time-dependent manner. This enhancement was attenuated by isohelenin. Conclusions and Clinical Relevance - Data suggested that a nuclear factor κB-based transcriptional mechanism is involved in induction of MUC5AC expression by tumor necrosis factor-α. Understanding the molecular mechanism of cytokine-enhanced MUC5AC expression in horses may lead to better treatment options and understanding of the pathogenesis of equine pulmonary diseases.

Original languageEnglish (US)
Pages (from-to)690-696
Number of pages7
JournalAmerican Journal of Veterinary Research
Volume71
Issue number6
DOIs
StatePublished - Jun 2010

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Horses
epithelial cells
Up-Regulation
Epithelial Cells
horses
cytokines
Cytokines
tumor necrosis factors
respiratory tract diseases
Tumor Necrosis Factor-alpha
Horse Diseases
Respiratory Tract Diseases
B-DNA
Messenger RNA
Glyceraldehyde-3-Phosphate Dehydrogenases
Euthanasia
Serum-Free Culture Media
trachea (vertebrates)
Mucins
glyceraldehyde-3-phosphate dehydrogenase

ASJC Scopus subject areas

  • veterinary(all)

Cite this

Evaluation of MUC5AC expression and upregulation in airway epithelial cells of horses. / Oslund, Karen L.; Adamson, Grete; Wu, Reen.

In: American Journal of Veterinary Research, Vol. 71, No. 6, 06.2010, p. 690-696.

Research output: Contribution to journalArticle

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