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Muc5b is the major polymeric mucin in mucus from thoroughbred horses with and without airway mucus accumulation.

Rousseau K, Cardwell JM, Humphrey E, Newton R, Knight D, Clegg P, Thornton DJ - PLoS ONE (2011)

Bottom Line: Immunohistochemical staining of normal equine trachea showed that Muc5ac and Muc5b are produced by cells in the submucosal glands, as well as surface epithelial goblet cells.The change in amount of Muc5b and Muc5ac indicates that Muc5b remains the most abundant mucin in mucus.In summary, we have developed mucin specific polyclonal antibodies, which have allowed us to show that there is a significant increase in Muc5b and Muc5ac in mucus accumulated in equine airways and these increases correlated with the numbers of bacteria.

View Article: PubMed Central - PubMed

Affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom.

ABSTRACT
Mucus accumulation is a feature of inflammatory airway disease in the horse and has been associated with reduced performance in racehorses. In this study, we have analysed the two major airways gel-forming mucins Muc5b and Muc5ac in respect of their site of synthesis, their biochemical properties, and their amounts in mucus from healthy horses and from horses with signs of airway mucus accumulation. Polyclonal antisera directed against equine Muc5b and Muc5ac were raised and characterised. Immunohistochemical staining of normal equine trachea showed that Muc5ac and Muc5b are produced by cells in the submucosal glands, as well as surface epithelial goblet cells. Western blotting after agarose gel electrophoresis of airway mucus from healthy horses, and horses with mucus accumulation, was used to determine the amounts of these two mucins in tracheal wash samples. The results showed that in healthy horses Muc5b was the predominant mucin with small amounts of Muc5ac. The amounts of Muc5b and Muc5ac were both dramatically increased in samples collected from horses with high mucus scores as determined visually at the time of endoscopy and that this increase also correlated with increase number of bacteria present in the sample. The change in amount of Muc5b and Muc5ac indicates that Muc5b remains the most abundant mucin in mucus. In summary, we have developed mucin specific polyclonal antibodies, which have allowed us to show that there is a significant increase in Muc5b and Muc5ac in mucus accumulated in equine airways and these increases correlated with the numbers of bacteria.

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Related in: MedlinePlus

Determination of the sensitivity of the MANeq5b-I and MANeq5ac-I antisera.Reduced and carboxymethylated mucin standard solutions, prepared from saliva (Muc5b 10-0.02 ng) and from equine stomach mucus (Muc5ac, 1480-2.9 ng), were electrophoresed on 0.7% agarose gels. Western blots of the gels were probed with MANeq5b-I (inset in panel A) and MANeq5ac-I (inset in panel B), the intensity of the staining was measured by reflectance densitometry and is plotted against the amount of mucin.
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pone-0019678-g009: Determination of the sensitivity of the MANeq5b-I and MANeq5ac-I antisera.Reduced and carboxymethylated mucin standard solutions, prepared from saliva (Muc5b 10-0.02 ng) and from equine stomach mucus (Muc5ac, 1480-2.9 ng), were electrophoresed on 0.7% agarose gels. Western blots of the gels were probed with MANeq5b-I (inset in panel A) and MANeq5ac-I (inset in panel B), the intensity of the staining was measured by reflectance densitometry and is plotted against the amount of mucin.

Mentions: A range of amounts of the two-mucin standard solutions was electrophoresed and Western blots of the gels were probed with either MANeq5ac-I or MANeq5b-I. The response curves generated show that eqMuc5ac-I recognises substantially lower levels of mucin than the eqMuc5b-I (Figure 9). MANeq5b-I antibody detected between 0.1 to 10 µg of Muc5b while MANeq5b-I detected between 10 to 1000 ng of Muc5ac (Figure 9). A retrospective analysis of the data generated above showed that Muc5b is the major mucin in the airway mucus, and that although the amounts of both mucins could be increased by more than 10-fold in tracheal wash samples from horses with mucus accumulation, Muc5b remained the most abundant mucin in the vast majority of samples analysed.


Muc5b is the major polymeric mucin in mucus from thoroughbred horses with and without airway mucus accumulation.

Rousseau K, Cardwell JM, Humphrey E, Newton R, Knight D, Clegg P, Thornton DJ - PLoS ONE (2011)

Determination of the sensitivity of the MANeq5b-I and MANeq5ac-I antisera.Reduced and carboxymethylated mucin standard solutions, prepared from saliva (Muc5b 10-0.02 ng) and from equine stomach mucus (Muc5ac, 1480-2.9 ng), were electrophoresed on 0.7% agarose gels. Western blots of the gels were probed with MANeq5b-I (inset in panel A) and MANeq5ac-I (inset in panel B), the intensity of the staining was measured by reflectance densitometry and is plotted against the amount of mucin.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3094342&req=5

pone-0019678-g009: Determination of the sensitivity of the MANeq5b-I and MANeq5ac-I antisera.Reduced and carboxymethylated mucin standard solutions, prepared from saliva (Muc5b 10-0.02 ng) and from equine stomach mucus (Muc5ac, 1480-2.9 ng), were electrophoresed on 0.7% agarose gels. Western blots of the gels were probed with MANeq5b-I (inset in panel A) and MANeq5ac-I (inset in panel B), the intensity of the staining was measured by reflectance densitometry and is plotted against the amount of mucin.
Mentions: A range of amounts of the two-mucin standard solutions was electrophoresed and Western blots of the gels were probed with either MANeq5ac-I or MANeq5b-I. The response curves generated show that eqMuc5ac-I recognises substantially lower levels of mucin than the eqMuc5b-I (Figure 9). MANeq5b-I antibody detected between 0.1 to 10 µg of Muc5b while MANeq5b-I detected between 10 to 1000 ng of Muc5ac (Figure 9). A retrospective analysis of the data generated above showed that Muc5b is the major mucin in the airway mucus, and that although the amounts of both mucins could be increased by more than 10-fold in tracheal wash samples from horses with mucus accumulation, Muc5b remained the most abundant mucin in the vast majority of samples analysed.

Bottom Line: Immunohistochemical staining of normal equine trachea showed that Muc5ac and Muc5b are produced by cells in the submucosal glands, as well as surface epithelial goblet cells.The change in amount of Muc5b and Muc5ac indicates that Muc5b remains the most abundant mucin in mucus.In summary, we have developed mucin specific polyclonal antibodies, which have allowed us to show that there is a significant increase in Muc5b and Muc5ac in mucus accumulated in equine airways and these increases correlated with the numbers of bacteria.

View Article: PubMed Central - PubMed

Affiliation: Wellcome Trust Centre for Cell-Matrix Research, Faculty of Life Sciences, University of Manchester, Manchester, United Kingdom.

ABSTRACT
Mucus accumulation is a feature of inflammatory airway disease in the horse and has been associated with reduced performance in racehorses. In this study, we have analysed the two major airways gel-forming mucins Muc5b and Muc5ac in respect of their site of synthesis, their biochemical properties, and their amounts in mucus from healthy horses and from horses with signs of airway mucus accumulation. Polyclonal antisera directed against equine Muc5b and Muc5ac were raised and characterised. Immunohistochemical staining of normal equine trachea showed that Muc5ac and Muc5b are produced by cells in the submucosal glands, as well as surface epithelial goblet cells. Western blotting after agarose gel electrophoresis of airway mucus from healthy horses, and horses with mucus accumulation, was used to determine the amounts of these two mucins in tracheal wash samples. The results showed that in healthy horses Muc5b was the predominant mucin with small amounts of Muc5ac. The amounts of Muc5b and Muc5ac were both dramatically increased in samples collected from horses with high mucus scores as determined visually at the time of endoscopy and that this increase also correlated with increase number of bacteria present in the sample. The change in amount of Muc5b and Muc5ac indicates that Muc5b remains the most abundant mucin in mucus. In summary, we have developed mucin specific polyclonal antibodies, which have allowed us to show that there is a significant increase in Muc5b and Muc5ac in mucus accumulated in equine airways and these increases correlated with the numbers of bacteria.

Show MeSH
Related in: MedlinePlus