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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

Sedimentation analysis of Muc5ac and Muc5b.Unreduced (panel A) and reduced mucins (panel B) were subjected to rate-zonal centrifugation on 6-8 M guanidinium chloride gradients. Fractions were analysed for glycoprotein content by PAS-staining (black triangles), and for reactivity with MANeq5b-I (black circles) and MANeq5ac-I (open circles). Fractions 1 and 24 represent the top and bottom of the gradient, respectively.
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pone-0019678-g004: Sedimentation analysis of Muc5ac and Muc5b.Unreduced (panel A) and reduced mucins (panel B) were subjected to rate-zonal centrifugation on 6-8 M guanidinium chloride gradients. Fractions were analysed for glycoprotein content by PAS-staining (black triangles), and for reactivity with MANeq5b-I (black circles) and MANeq5ac-I (open circles). Fractions 1 and 24 represent the top and bottom of the gradient, respectively.

Mentions: To analyse the size distributions of Muc5ac and Muc5b mucins, airway mucus (pooled from 4 horses with airway mucus accumulation) was subjected to rate-zonal centrifugation, before and after treatment with a reducing agent (10 mM DTT). The unreduced Muc5ac and Muc5b mucins were characterized by a broad range of sedimentation rates, characteristic of a polydisperse distribution of mucins (Figure 4A). After reduction, the mucins exhibited slower sedimentation rates. Interestingly, there was a separation of the PAS-reactivity from a slower sedimenting population of molecules that was highly reactive with the mucin probes (Figure 4B).


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)

Sedimentation analysis of Muc5ac and Muc5b.Unreduced (panel A) and reduced mucins (panel B) were subjected to rate-zonal centrifugation on 6-8 M guanidinium chloride gradients. Fractions were analysed for glycoprotein content by PAS-staining (black triangles), and for reactivity with MANeq5b-I (black circles) and MANeq5ac-I (open circles). Fractions 1 and 24 represent the top and bottom of the gradient, respectively.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0019678-g004: Sedimentation analysis of Muc5ac and Muc5b.Unreduced (panel A) and reduced mucins (panel B) were subjected to rate-zonal centrifugation on 6-8 M guanidinium chloride gradients. Fractions were analysed for glycoprotein content by PAS-staining (black triangles), and for reactivity with MANeq5b-I (black circles) and MANeq5ac-I (open circles). Fractions 1 and 24 represent the top and bottom of the gradient, respectively.
Mentions: To analyse the size distributions of Muc5ac and Muc5b mucins, airway mucus (pooled from 4 horses with airway mucus accumulation) was subjected to rate-zonal centrifugation, before and after treatment with a reducing agent (10 mM DTT). The unreduced Muc5ac and Muc5b mucins were characterized by a broad range of sedimentation rates, characteristic of a polydisperse distribution of mucins (Figure 4A). After reduction, the mucins exhibited slower sedimentation rates. Interestingly, there was a separation of the PAS-reactivity from a slower sedimenting population of molecules that was highly reactive with the mucin probes (Figure 4B).

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