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Assessment of Intracellular Mucin Content In Vivo

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Book cover Mucins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 842))

Abstract

Airway mucus presents a first line of defense against inhaled materials. It also, however, is a significant pathological contributor to chronic lung diseases, such as asthma, cystic fibrosis, and chronic obstructive pulmonary disease. Thus, gaining a better understanding of the mechanisms of mucus production and secretion is an important goal for improving respiratory health. Mucins, the chief glycoprotein components of airway mucus, are very large polymeric glycoproteins, and measuring their production and secretion in experimental animals presents significant technical challenges. Over the past several years, we have developed assays for accurately quantifying mucin production and secretion using histological and biochemical assays. These methods are described here.

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References

  1. Davis, C. W. and Dickey, B. F. (2008) Regulated airway goblet cell mucin secretion. Annu. Rev. Physiol. 70, 487–512.

    Google Scholar 

  2. Evans, C. M. and Koo, J. S. (2009) Airway mucus: the good, the bad, the sticky. Pharmacol. Ther. 121, 332–348.

    Google Scholar 

  3. Fahy, J. V. and Dickey, B. F. (2010) Airway mucus function and dysfunction N. Engl. J. Med. 363, 2233–2247.

    Google Scholar 

  4. Young, H. W., Williams, O. W., Chandra, D., Bellinghausen, L. K., Perez, G., Suarez, A., Tuvim, M. J., Roy, M. G., Alexander, S. N., Moghaddam, S. J., Adachi, R., Blackburn, M. R., Dickey, B. F., and Evans, C. M. (2007) Central role of Muc5ac expression in mucous metaplasia and its regulation by conserved 5’ elements Am J Respir Cell Mol Biol. 37, 273–290.

    Google Scholar 

  5. Chen, Y., Zhao, Y. H., and Wu, R. (2001) In Silico Cloning of Mouse Muc5b Gene and Upregulation of Its Expression in Mouse Asthma Model. Am. J Respir. Crit Care Med 164, 1059–1066.

    Google Scholar 

  6. Zhu, Y., Ehre, C., Abdullah, L. H., Sheehan, J. K., Roy, M., Evans, C. M., Dickey, B. F., and Davis, C. W. Munc13-2−/− baseline secretion defect reveals source of oligomeric mucins in mouse airways. (2008) J Physiol. 586, 1977–1992.

    Google Scholar 

  7. Nguyen, L. P., Omoluabi, O., Parra, S., Frieske, J. M., Clement, C., mmar-Aouchiche, Z., Ho, S. B., Ehre, C., Kesimer, M., Knoll, B. J., Tuvim, M. J., Dickey, B. F., and Bond, R. A. (2008) Chronic exposure to beta-blockers attenuates inflammation and mucin content in a murine asthma model. Am J Respir Cell Mol. Biol. 38, 256–262.

    Google Scholar 

  8. Zudhi Alimam, M., Piazza, F. M., Selby, D. M., Letwin, N., Huang, L., and Rose, M. C. (2000) Muc-5/5ac mucin messenger RNA and protein expression is a marker of goblet cell metaplasia in murine airways. Am. J. Respir. Cell Mol. Biol. 22, 253–260.

    Google Scholar 

  9. Evans, C. M., Williams, O. W., Tuvim, M. J., Nigam, R., Mixides, G. P., Blackburn, M. R., DeMayo, F. J., Burns, A. R., Smith, C., Reynolds, S. D., Stripp, B. R., and Dickey, B. F. (2004) Mucin is produced by clara cells in the proximal airways of antigen-challenged mice. Am J. Respir. Cell Mol. Biol. 31, 382–394.

    Google Scholar 

  10. Davis, C. W. and Lazarowski, E. (2008) Coupling of airway ciliary activity and mucin secretion to mechanical stresses by purinergic signaling. Respir. Physiol Neurobiol. 163, 208–213.

    Google Scholar 

  11. Singer, M., Martin, L. D., Vargaftig, B. B., Park, J., Gruber, A. D., Li, Y., and Adler, K. B. (2004) A MARCKS-related peptide blocks mucus hypersecretion in a mouse model of asthma. Nat. Med. 10, 193–196.

    Google Scholar 

  12. Tuvim, M. J., Mospan, A. R., Burns, K. A., Chua, M., Mohler, P. J., Melicoff, E., Adachi, R., mmar-Aouchiche, Z., Davis, C. W., and Dickey, B. F. (2009) Synaptotagmin 2 couples mucin granule exocytosis to Ca2+ signaling from endoplasmic reticulum. J Biol. Chem. 284, 9781–9787.

    Google Scholar 

  13. Grunig, G., Warnock, M., Wakil, A. E., Venkayya, R., Brombacher, F., Rennick, D. M., Sheppard, D., Mohrs, M., Donaldson, D. D., Locksley, R. M., and Corry, D. B. (1998) Requirement for IL-13 independently of IL-4 in experimental asthma. Science 282, 2261–2263.

    Google Scholar 

  14. Lappalainen, U., Whitsett, J. A., Wert, S. E., Tichelaar, J. W., and Bry, K. (2005) Interleukin-1beta causes pulmonary inflammation, emphysema, and airway remodeling in the adult murine lung. Am J Respir Cell Mol. Biol. 32, 311–318.

    Google Scholar 

  15. Tomkinson, A., Cieslewicz, G., Duez, C., Larson, K. A., Lee, J. J., and Gelfand, E. W. (2001) Temporal association between airway hyperresponsiveness and airway eosinophilia in ovalbumin-sensitized mice. Am J Respir Crit Care Med. 163, 721–730.

    Google Scholar 

  16. Trifilieff, A., Ahmed, E., and Bertrand, C. (2000). Time course of inflammatory and remodeling events in a murine model of asthma: effect of steroid treatment. Am. J. Physiol Lung Cell Mol. Physiol 279, L1120–L1128.

    Google Scholar 

  17. Walter, M. J., Morton, J. D., Kajiwara, N., Agapov, E., and Holtzman, M. J. (2002) Viral induction of a chronic asthma phenotype and genetic segregation from the acute response. J. Clin. Invest 110, 165–175.

    Google Scholar 

  18. Wills-Karp, M., Luyimbazi, J., Xu, X., Schofield, B., Neben, T. Y., Karp, C. L., and Donaldson, D. D. (1998) Interleukin-13: central mediator of allergic asthma. Science 282, 2258–2261.

    Google Scholar 

  19. Zheng, T., Zhu, Z., Wang, Z., Homer, R. J., Ma, B., Riese, R. J., Chapman, H. A., Shapiro, S. D., and Elias, J. A. (2000) Inducible targeting of IL-13 to the adult lung causes matrix metalloproteinase- and cathepsin-dependent emphysema. J. Clin. Invest 106, 1081–1093.

    Google Scholar 

  20. Carlstedt, I., Lindgren, H., Sheehan, J. K., Ulmsten, U., and Wingerup, L. (1983) Isolation and characterization of human cervical-mucus glycoproteins. Biochem. J. 211, 13–22.

    Google Scholar 

  21. Aksoy, N., Thornton, D. J., Corfield, A., Paraskeva, C., and Sheehan, J. K. (1999) A study of the intracellular and secreted forms of the MUC2 mucin from the PC/AA intestinal cell line. Glycobiology 9, 739–746.

    Google Scholar 

  22. Thornton, D. J., Rousseau, K., and McGuckin, M. A. (2007) Structure and function of the polymeric mucins in airways mucus. Annu. Rev Physiol. 70, 459–86.

    Google Scholar 

  23. Reader, J. R., Tepper, J. S., Schelegle, E. S., Aldrich, M. C., Putney, L. F., Pfeiffer, J. W., and Hyde, D. M. (2003) Pathogenesis of mucous cell metaplasia in a murine asthma model. Am. J. Pathol. 162, 2069–2078.

    Google Scholar 

  24. Weibel, E. R. (1979) Sterological Methods Academic Press Inc. Ltd., London.

    Google Scholar 

  25. Harkema, J. R., Plopper, C. G., Hyde, D. M., and St George, J. A. (1987) Regional differences in quantities of histochemically detectable mucosubstances in nasal, paranasal, and nasopharyngeal epithelium of the bonnet monkey. J. Histochem. Cytochem. 35, 279–286.

    Google Scholar 

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Acknowledgments

This work was supported by NIH grants HL080396 (C.M. Evans), HL094848 (B.F. Dickey), and HL097000 (B.F. Dickey), American Heart Association Grant 10GRNT4200070 (C.M. Evans), and Cystic Fibrosis Foundation grant 08GO (B.F. Dickey). The authors thank C. William Davis for instruction in the performance of vacuum blotting of mucins.

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Authors and Affiliations

  1. Department of Pulmonary Medicine, The University of Texas M.D. Anderson Cancer Center, Houston, TX, USA

    Lucia Piccotti, Burton F. Dickey & Christopher M. Evans

Authors
  1. Lucia Piccotti
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  2. Burton F. Dickey
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  3. Christopher M. Evans
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Corresponding author

Correspondence to Christopher M. Evans .

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Editors and Affiliations

  1. Mucosal Diseases Program, Mater Medical Research Institute, Aubigny Place, South Brisbane, 4101, Queensland, Australia

    Michael A. McGuckin

  2. Fac. Life Sciences, Wellcome Trust Centre for, University of Manchester, Oxford Road, Manchester, M13 9PT, United Kingdom

    David J. Thornton

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Piccotti, L., Dickey, B.F., Evans, C.M. (2012). Assessment of Intracellular Mucin Content In Vivo. In: McGuckin, M., Thornton, D. (eds) Mucins. Methods in Molecular Biology, vol 842. Humana Press. https://doi.org/10.1007/978-1-61779-513-8_17

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  • DOI: https://doi.org/10.1007/978-1-61779-513-8_17

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-512-1

  • Online ISBN: 978-1-61779-513-8

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