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
Davis, C. W. and Dickey, B. F. (2008) Regulated airway goblet cell mucin secretion. Annu. Rev. Physiol. 70, 487–512.
Evans, C. M. and Koo, J. S. (2009) Airway mucus: the good, the bad, the sticky. Pharmacol. Ther. 121, 332–348.
Fahy, J. V. and Dickey, B. F. (2010) Airway mucus function and dysfunction N. Engl. J. Med. 363, 2233–2247.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Weibel, E. R. (1979) Sterological Methods Academic Press Inc. Ltd., London.
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.
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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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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