Quantification of mucin gene expression in tracheobronchial epithelium of healthy dogs and dogs with chronic bronchitis.
OBJECTIVE
To develop a real-time PCR assay for the quantification of mucin gene expression in tracheobronchial brushing specimens from dogs and compare mucin gene expression in specimens from dogs with naturally occurring chronic bronchitis with that in specimens from healthy dogs.


METHODS
7 healthy dogs and 5 dogs with chronic bronchitis.


METHODS
Primers that were designed to span the predicted intron-exon boundaries of a canine MUC5AC-like gene were used to develop a real-time PCR assay for quantification of expression of that gene. Total mRNA was isolated from tracheobronchial brushing specimens obtained from dogs with and without bronchitis during anesthesia; MUC5AC-like gene expression in those samples was quantified by use of the real-time PCR assay.


RESULTS
The PCR assay was sensitive and specific for the target sequence, the predicted amino acid sequence of which had greatest homology with human, porcine, and rat MUC5AC. The assay was able to quantify the target over a wide dynamic range. Dogs with chronic bronchitis had a 3.0-fold increase in the quantity of MUC5AC-like mRNA, compared with healthy dogs.


CONCLUSIONS
The ability to measure mucin gene expression from tracheobronchial brushing specimens collected from client-owned dogs during routine bronchoscopy should prove to be a useful tool for the study of bronchitis in dogs and expand the usefulness of airway inflammation in dogs as a model for bronchitis in humans.
Hawkins
Eleanor C
EC
Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27606, USA.
Birkenheuer
Adam J
AJ
Marr
Henry S
HS
Rogala
Allison R
AR
Large
Edward E
EE
Adler
Kenneth B
KB
D000818:Animals; D029481:Bronchitis, Chronic / Q000378:metabolism / Q000662:veterinary*; D017931:DNA Primers; D004283:Dog Diseases / Q000378:metabolism*; D004285:Dogs; D005069:Evaluation Studies as Topic; D009077:Mucins / Q000378:metabolism*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012333:RNA, Messenger / Q000378:metabolism*
D017931:DNA Primers; D009077:Mucins; D012333:RNA, Messenger