Transcriptional changes associated with recurrent airway obstruction in affected and unaffected horses

Changaram S. Venugopal Equine Health Studies Program, Department of Veterinary Clinical Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA 70803.

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Luiz Claudio N. Mendes Department of Clinics, Surgery and Animal Reproduction, Sao Paulo State University, UNESP, R Clovis Pestana 793, Aracatuba, SP 16050-680, Brazil.

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Juliana R. Peiró Department of Clinics, Surgery and Animal Reproduction, Sao Paulo State University, UNESP, R Clovis Pestana 793, Aracatuba, SP 16050-680, Brazil.

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Susan S. Laborde Equine Health Studies Program, Department of Veterinary Clinical Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA 70803.

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Ashley M. Stokes Equine Health Studies Program, Department of Veterinary Clinical Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA 70803.

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Rustin M. Moore Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, Columbus, OH 43210-1089.

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Abstract

Objective—To identify differentially expressed genes in pulmonary tissues of horses affected with summer pasture-associated obstructive pulmonary disease (SPAOPD), which is a form of recurrent airway obstruction (RAO), compared with those of unaffected horses.

Animals—6 horses with SPAOPD-RAO and 6 unaffected (healthy) horses.

Procedures—Horses were assigned to 2 groups on the basis of medical history, clinical score, and transpleural pressure. Total RNA from each of the 5 lung lobes of each of the 6 SPAOPD-RAO–affected horses was extracted and pooled. Similarly, total RNA from unaffected horses was pooled. Differential display (DD) PCR assay was performed, and differentially expressed bands were purified and cloned into a plasmid vector. Plasmids were extracted from recombinant colonies, and purified DNA was sequenced. Genes of interest for RAO pathogenesis were identified. Real-time PCR assay was performed to confirm findings for the DD PCR assay.

Results—18 differentially expressed genes (17 upregulated and 1 downregulated) were identified. Three genes of particular interest were found to be altered (2 upregulated and 1 downregulated) in horses with SPAOPD-RAO by use of real-time PCR assay, and these findings matched the differential expression found by use of the DD PCR assay.

Conclusions and Clinical Relevance—SPAOPD-RAO in horses is a multifactorial, complex disease involving several genes. Upregulated genes, particularly β2-microglobulin, and the downregulated secretoglobin gene can serve as marker genes that may help to identify SPAOPD-RAO at an early age.

Abstract

Objective—To identify differentially expressed genes in pulmonary tissues of horses affected with summer pasture-associated obstructive pulmonary disease (SPAOPD), which is a form of recurrent airway obstruction (RAO), compared with those of unaffected horses.

Animals—6 horses with SPAOPD-RAO and 6 unaffected (healthy) horses.

Procedures—Horses were assigned to 2 groups on the basis of medical history, clinical score, and transpleural pressure. Total RNA from each of the 5 lung lobes of each of the 6 SPAOPD-RAO–affected horses was extracted and pooled. Similarly, total RNA from unaffected horses was pooled. Differential display (DD) PCR assay was performed, and differentially expressed bands were purified and cloned into a plasmid vector. Plasmids were extracted from recombinant colonies, and purified DNA was sequenced. Genes of interest for RAO pathogenesis were identified. Real-time PCR assay was performed to confirm findings for the DD PCR assay.

Results—18 differentially expressed genes (17 upregulated and 1 downregulated) were identified. Three genes of particular interest were found to be altered (2 upregulated and 1 downregulated) in horses with SPAOPD-RAO by use of real-time PCR assay, and these findings matched the differential expression found by use of the DD PCR assay.

Conclusions and Clinical Relevance—SPAOPD-RAO in horses is a multifactorial, complex disease involving several genes. Upregulated genes, particularly β2-microglobulin, and the downregulated secretoglobin gene can serve as marker genes that may help to identify SPAOPD-RAO at an early age.

Contributor Notes

Dr. Stokes' present address is the Department of Human Nutrition, Food and Animal Sciences, College of Tropical Agriculture and Human Resources, University of Hawaii, Honolulu, HI 96822.

Supported by the Equine Health Studies Program, School of Veterinary Medicine, Louisiana State University.

Dr. Mendes was supported by a scholarship from CNPq – Brazil (201044/2005-6).

Presented in abstract form at the Experimental Biology Annual Meeting, Washington, DC, April–May 2007.

Address correspondence to Dr. Venugopal (cvenugopal@vetmed.lsu.edu).
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