Abstract
Chronic otitis media (COM), e.g. “glue” ear is characterized by middle ear effusion and conductive hearing loss. Although mucous glycoproteins (mucins), which contribute to increased effusion viscosity, have been analyzed in ear tissue specimens, no studies have been reported that characterize the molecular identity of secreted mucin proteins present in actual middle ear fluid. For this study, effusions from children with COM undergoing myringotomy at Children's National Medical Center, Washington, DC were collected. These were solubilized and gel fractionated, and the protein content was identified using a liquid chromatography tandem mass spectrometry (LC-MS/MS) proteomics approach. Western blot analyses with mucin specific antibodies and densitometry were performed to validate the mass spectrometry findings. LC-MS/MS results identified mucin MUC5B by >26 unique peptides in six of six middle ear effusion samples, whereas mucin MUC5AC was only identified in one of six middle ear effusions. These findings were validated by Western blot performed on the same six and on an additional 11 separate samples where densitometry revealed on average a 6.4-fold increased signal in MUC5B when compared with MUC5AC (p = 0.0009). In summary, although both MUC5AC and MUC5B mucins are detected in middle ear effusions, MUC5B seems to be predominant mucin present in COM secretions.
Similar content being viewed by others
Log in or create a free account to read this content
Gain free access to this article, as well as selected content from this journal and more on nature.com
or
Abbreviations
- COM:
-
Chronic otitis media
- LC-MS/MS:
-
liquid chromatography tandem mass spectrometry
- MS:
-
mass spectrometry
- MS/MS:
-
tandem mass spectrometry
- MUC:
-
mucin
- OM:
-
otitis media
References
Cherry DK, Woodwell DA 2002 National Ambulatory Medical Care Survey: 2000 summary. Adv Data 328: 1–32
Rosenfeld RM, Casselbrant ML, Hannley MT 2001 Implications of the AHRQ evidence report on acute otitis media. Otolaryngol Head Neck Surg 125: 440–448; discussion 439
McIntosh ED, Conway P, Willingham J, Lloyd A 2003 The cost-burden of paediatric pneumococcal disease in the UK and the potential cost-effectiveness of prevention using 7-valent pneumococcal conjugate vaccine. Vaccine 21: 2564–2572
Chung MH, Choi JY, Lee WS, Kim HN, Yoon JH 2002 Compositional difference in middle ear effusion: mucous versus serous. Laryngoscope 112: 152–155
1969 Glue ear. BMJ 4: 578
Klein JO 2000 The burden of otitis media. Vaccine 19: S2–S8
Teele DW, Klein JO, Rosner BA 1984 Otitis media with effusion during the first three years of life and development of speech and language. Pediatrics 74: 282–287
Brown DT, Litt M, Potsic WP 1985 A study of mucus glycoproteins in secretory otitis media. Arch Otolaryngol 111: 688–695
Carrie S, Hutton DA, Birchall JP, Green GG, Pearson JP 1992 Otitis media with effusion: components which contribute to the viscous properties. Acta Otolaryngol 112: 504–511
Lous J, Burton MJ, Felding JU, Ovesen T, Rovers MM, Williamson I 2005 Grommets (ventilation tubes) for hearing loss associated with otitis media with effusion in children. Cochrane Database Syst Rev CD001801
Bluestone CD, Stool SE, Kenna MA 1996 Pediatric Otolaryngology. Saunders, Philadelphia
Kerschner JE 2007 Mucin gene expression in human middle ear epithelium. Laryngoscope 117: 1666–1676
Giebink GS, Le CT, Paparella MM 1982 Epidemiology of otitis media with effusion in children. Arch Otolaryngol 108: 563–566
Rose MC 1992 Mucins: structure, function, and role in pulmonary diseases. Am J Physiol 263: L413–L429
Ali MS, Pearson JP 2007 Upper airway mucin gene expression: a review. Laryngoscope 117: 932–938
Rose MC, Voynow JA 2006 Respiratory tract mucin genes and mucin glycoproteins in health and disease. Physiol Rev 86: 245–278
Gendler SJ, Spicer AP 1995 Epithelial mucin genes. Annu Rev Physiol 57: 607–634
Hovenberg HW, Davies JR, Herrmann A, Linden CJ, Carlstedt I 1996 MUC5AC, but not MUC2, is a prominent mucin in respiratory secretions. Glycoconj J 13: 839–847
Hathout Y 2007 Approaches to the study of the cell secretome. Expert Rev Proteomics 4: 239–248
Ke E, Patel BB, Liu T, Li XM, Haluszka O, Hoffman JP, Ehya H, Young NA, Watson JC, Weinberg DS, Nguyen MT, Cohen SJ, Meropol NJ, Litwin S, Tokar JL, Yeung AT 2009 Proteomic analyses of pancreatic cyst fluids. Pancreas 38: e33–e42
Rousseau K, Kirkham S, Johnson L, Fitzpatrick B, Howard M, Adams EJ, Rogers DF, Knight D, Clegg P, Thornton DJ 2008 Proteomic analysis of polymeric salivary mucins: no evidence for MUC19 in human saliva. Biochem J 413: 545–552
Rousseau K, Kirkham S, McKane S, Newton R, Clegg P, Thornton DJ 2007 Muc5b and Muc5ac are the major oligomeric mucins in equine airway mucus. Am J Physiol Lung Cell Mol Physiol 292: L1396–L1404
Andersch-Björkman Y, Thomsson KA, Holmén Larsson JM, Ekerhovd E, Hansson GC 2007 Large scale identification of proteins, mucins, and their O-glycosylation in the endocervical mucus during the menstrual cycle. Mol Cell Proteomics 6: 708–716
Walz A, Stuhler K, Wattenberg A, Hawranke E, Meyer HE, Schmalz G, Bluggel M, Ruhl S 2006 Proteome analysis of glandular parotid and submandibular-sublingual saliva in comparison to whole human saliva by two-dimensional gel electrophoresis. Proteomics 6: 1631–1639
Berger JT, Voynow JA, Peters KW, Rose MC 1999 Respiratory carcinoma cell lines. MUC genes and glycoconjugates. Am J Respir Cell Mol Biol 20: 500–510
Troxler RF, Offner GD, Zhang F, Iontcheva I, Oppenheim FG 1995 Molecular cloning of a novel high molecular weight mucin (MG1) from human sublingual gland. Biochem Biophys Res Commun 217: 1112–1119
Lin J, Tsuprun V, Kawano H, Paparella MM, Zhang Z, Anway R, Ho SB 2001 Characterization of mucins in human middle ear and Eustachian tube. Am J Physiol Lung Cell Mol Physiol 280: L1157–L1167
Lin J, Tsuboi Y, Rimell F, Liu G, Toyama K, Kawano H, Paparella MM, Ho SB 2003 Expression of mucins in mucoid otitis media. J Assoc Res Otolaryngol 4: 384–393
Choi JY, Kim CH, Lee WS, Kim HN, Song KS, Yoon JH 2002 Ciliary and secretory differentiation of normal human middle ear epithelial cells. Acta Otolaryngol 122: 270–275
Ubell ML, Kerschner JE, Wackym PA, Burrows A 2008 MUC2 expression in human middle ear epithelium of patients with otitis media. Arch Otolaryngol Head Neck Surg 134: 39–44
Takeuchi K, Yagawa M, Ishinaga H, Kishioka C, Harada T, Majima Y 2003 Mucin gene expression in the effusions of otitis media with effusion. Int J Pediatr Otorhinolaryngol 67: 53–58
Jansen RC, Nap JP, Mlynarova L 2002 Errors in genomics and proteomics. Nat Biotechnol 20: 19
Kussmann M, Raymond F, Affolter M 2006 OMICS-driven biomarker discovery in nutrition and health. J Biotechnol 124: 758–787
Schousboe LP, Rasmussen LM, Ovesen T 2001 Induction of mucin and adhesion molecules in middle ear mucosa. Acta Otolaryngol 121: 596–601
Elsheikh MN, Mahfouz ME 2006 Up-regulation of MUC5AC and MUC5B mucin genes in nasopharyngeal respiratory mucosa and selective up-regulation of MUC5B in middle ear in pediatric otitis media with effusion. Laryngoscope 116: 365–369
Reid CJ, Gould S, Harris A 1997 Developmental expression of mucin genes in the human respiratory tract. Am J Respir Cell Mol Biol 17: 592–598
Audie JP, Janin A, Porchet N, Copin MC, Gosselin B, Aubert JP 1993 Expression of human mucin genes in respiratory, digestive, and reproductive tracts ascertained by in situ hybridization. J Histochem Cytochem 41: 1479–1485
Preciado D, Lin J, Wuertz B, Rose M 2008 Cigarette smoke activates NF kappa B and induces MUC5B expression in mouse middle ear cells. Laryngoscope 118: 464–471
Desseyn JL, Guyonnet-Duperat V, Porchet N, Aubert JP, Laine A 1997 Human mucin gene MUC5B, the 10.7-kb large central exon encodes various alternate subdomains resulting in a super-repeat. Structural evidence for a 11p15.5 gene family. J Biol Chem 272: 3168–3178
Keates AC, Nunes DP, Afdhal NH, Troxler RF, Offner GD 1997 Molecular cloning of a major human gall bladder mucin: complete C-terminal sequence and genomic organization of MUC5B. Biochem J 324: 295–303
Sheehan JK, Howard M, Richardson PS, Longwill T, Thornton DJ 1999 Physical characterization of a low-charge glycoform of the MUC5B mucin comprising the gel-phase of an asthmatic respiratory mucous plug. Biochem J 338: 507–513
Groneberg DA, Eynott PR, Lim S, Oates T, Wu R, Carlstedt I, Roberts P, McCann B, Nicholson AG, Harrison BD, Chung KF 2002 Expression of respiratory mucins in fatal status asthmaticus and mild asthma. Histopathology 40: 367–373
Kawano H, Paparella MM, Ho SB, Schachern PA, Morizono N, Le CT, Lin J 2000 Identification of MUC5B mucin gene in human middle ear with chronic otitis media. Laryngoscope 110: 668–673
Author information
Authors and Affiliations
Corresponding author
Additional information
Supported by NIH National Heart, Lung and Blood Institute, K12 Genomics of Lung Grant, HL090020-01. Intellectual and Developmental Disability Research Centers Core Grants 1P30HD40677 and 5R24HD050846 to the Center for Genetic Medicine at Children's National Medical Center, and an Avery Scholar Award from Children's National Medical Center [D.P.].
Supplemental digital content is available for this article. Direct URL citations appear in the printed text and are provided in the HTML and PDF versions of this article on the journal's Web site (www.pedresearch.org).
Rights and permissions
About this article
Cite this article
Preciado, D., Goyal, S., Rahimi, M. et al. MUC5B Is the Predominant Mucin Glycoprotein in Chronic Otitis Media Fluid. Pediatr Res 68, 231–236 (2010). https://doi.org/10.1203/PDR.0b013e3181eb2ecc
Received:
Accepted:
Issue date:
DOI: https://doi.org/10.1203/PDR.0b013e3181eb2ecc
This article is cited by
-
Inhibition of lipopolysaccharide-induced inflammation by trophoblast-conditioned medium and trophoblast-derived extracellular vesicles in human middle ear epithelial cells
Scientific Reports (2023)
-
Effect of vascular endothelial growth factor and its receptors in adult otitis media with effusion
European Archives of Oto-Rhino-Laryngology (2019)
-
Loss of the homeostatic protein BPIFA1, leads to exacerbation of otitis media severity in the Junbo mouse model
Scientific Reports (2018)
-
Characterization of mucoid and serous middle ear effusions from patients with chronic otitis media: implication of different biological mechanisms?
Pediatric Research (2018)
-
In vivo imaging of the Muc5b gel-forming mucin
Scientific Reports (2017)
