Abstract
The ocular surface, which is among the most accessible and vulnerable tissues in mammals, is protected by a complex tear film composed of lipid, aqueous and mucin layers. In spite of its importance, the molecular nature of the mucin contribution remains uncertain. Since membrane mucins have been implicated in the protection of other epithelia, we have analysed rat corneal and conjunctival tissues for sialomucin complex (SMC), a membrane mucin found at the apical epithelial cell surfaces in the airway and uterus. Using Northern and Western blot analyses, SMC expression was found in both ocular tissues, being particularly abundant in the cornea. In contrast with the other known membrane mucin, MUC1, SMC was localized more heavily towards the apical surface of the epithelial cells. SMC in ocular surface epithelia was produced in both soluble and membrane forms, the latter being found predominantly in the most superficial cells and at apical surfaces. The soluble form was found loosely adsorbed to apical cell surfaces, particularly of the cornea, as indicated by a mild rinsing protocol. Finally, the tear fluid contained substantial amounts of SMC. From these results, we propose a new model for tear mucin components in which SMC is expressed at the apical ocular surface in both membrane-bound and adsorbed soluble forms to provide a direct protective barrier. SMC secreted into the tear fluid may also participate in maintaining the stability of the preocular tear film by acting with other secreted mucins to determine the physical properties and protective behaviour of the tear film.
Full Text
The Full Text of this article is available as a PDF (301.6 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bobek L. A., Tsai H., Biesbrock A. R., Levine M. J. Molecular cloning, sequence, and specificity of expression of the gene encoding the low molecular weight human salivary mucin (MUC7). J Biol Chem. 1993 Sep 25;268(27):20563–20569. [PubMed] [Google Scholar]
- Carlstedt I., Sheehan J. K., Corfield A. P., Gallagher J. T. Mucous glycoproteins: a gel of a problem. Essays Biochem. 1985;20:40–76. [PubMed] [Google Scholar]
- Carraway K. L., Carraway C. A., Carraway K. L., 3rd Roles of ErbB-3 and ErbB-4 in the physiology and pathology of the mammary gland. J Mammary Gland Biol Neoplasia. 1997 Apr;2(2):187–198. doi: 10.1023/a:1026360032602. [DOI] [PubMed] [Google Scholar]
- Carraway K. L., Doss R. C., Huggins J. W., Chesnut R. W., Carraway C. A. Effects of cytoskeletal perturbant drugs on ecto 5'-nucleotidase, a concanavalin A receptor. J Cell Biol. 1979 Dec;83(3):529–543. doi: 10.1083/jcb.83.3.529. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chen H. B., Yamabayashi S., Ou B., Tanaka Y., Ohno S., Tsukahara S. Structure and composition of rat precorneal tear film. A study by an in vivo cryofixation. Invest Ophthalmol Vis Sci. 1997 Feb;38(2):381–387. [PubMed] [Google Scholar]
- Dilly P. N. Contribution of the epithelium to the stability of the tear film. Trans Ophthalmol Soc U K. 1985;104(Pt 4):381–389. [PubMed] [Google Scholar]
- Dilly P. N. On the nature and the role of the subsurface vesicles in the outer epithelial cells of the conjunctiva. Br J Ophthalmol. 1985 Jul;69(7):477–481. doi: 10.1136/bjo.69.7.477. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gendler S. J., Spicer A. P. Epithelial mucin genes. Annu Rev Physiol. 1995;57:607–634. doi: 10.1146/annurev.ph.57.030195.003135. [DOI] [PubMed] [Google Scholar]
- Gipson I. K., Yankauckas M., Spurr-Michaud S. J., Tisdale A. S., Rinehart W. Characteristics of a glycoprotein in the ocular surface glycocalyx. Invest Ophthalmol Vis Sci. 1992 Jan;33(1):218–227. [PubMed] [Google Scholar]
- Greiner J. V., Weidman T. A., Korb D. R., Allansmith M. R. Histochemical analysis of secretory vesicles in nongoblet conjunctival epithelial cells. Acta Ophthalmol (Copenh) 1985 Feb;63(1):89–92. doi: 10.1111/j.1755-3768.1985.tb05222.x. [DOI] [PubMed] [Google Scholar]
- Hicks S. J., Carrington S. D., Kaswan R. L., Adam S., Bara J., Corfield A. P. Demonstration of discrete secreted and membrane-bound ocular mucins in the dog. Exp Eye Res. 1997 Apr;64(4):597–607. doi: 10.1006/exer.1996.0246. [DOI] [PubMed] [Google Scholar]
- Holly F. J. Formation and rupture of the tear film. Exp Eye Res. 1973 May 10;15(5):515–525. doi: 10.1016/0014-4835(73)90064-x. [DOI] [PubMed] [Google Scholar]
- Huang A. J., Tseng S. C. Development of monoclonal antibodies to rabbit ocular mucin. Invest Ophthalmol Vis Sci. 1987 Sep;28(9):1483–1491. [PubMed] [Google Scholar]
- Hull S. R., Sheng Z., Vanderpuye O., David C., Carraway K. L. Isolation and partial characterization of ascites sialoglycoprotein-2 of the cell surface sialomucin complex of 13762 rat mammary adenocarcinoma cells. Biochem J. 1990 Jan 1;265(1):121–129. doi: 10.1042/bj2650121. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Inatomi T., Spurr-Michaud S., Tisdale A. S., Gipson I. K. Human corneal and conjunctival epithelia express MUC1 mucin. Invest Ophthalmol Vis Sci. 1995 Aug;36(9):1818–1827. [PubMed] [Google Scholar]
- Inatomi T., Spurr-Michaud S., Tisdale A. S., Zhan Q., Feldman S. T., Gipson I. K. Expression of secretory mucin genes by human conjunctival epithelia. Invest Ophthalmol Vis Sci. 1996 Jul;37(8):1684–1692. [PubMed] [Google Scholar]
- Komatsu M., Carraway C. A., Fregien N. L., Carraway K. L. Reversible disruption of cell-matrix and cell-cell interactions by overexpression of sialomucin complex. J Biol Chem. 1997 Dec 26;272(52):33245–33254. doi: 10.1074/jbc.272.52.33245. [DOI] [PubMed] [Google Scholar]
- Ligtenberg M. J., Kruijshaar L., Buijs F., van Meijer M., Litvinov S. V., Hilkens J. Cell-associated episialin is a complex containing two proteins derived from a common precursor. J Biol Chem. 1992 Mar 25;267(9):6171–6177. [PubMed] [Google Scholar]
- McNeer R. R., Carraway C. A., Fregien N. L., Carraway K. L. Characterization of the expression and steroid hormone control of sialomucin complex in the rat uterus: implications for uterine receptivity. J Cell Physiol. 1998 Jul;176(1):110–119. doi: 10.1002/(SICI)1097-4652(199807)176:1<110::AID-JCP13>3.0.CO;2-B. [DOI] [PubMed] [Google Scholar]
- McNeer R. R., Huang D., Fregien N. L., Carraway K. L. Sialomucin complex in the rat respiratory tract: a model for its role in epithelial protection. Biochem J. 1998 Mar 1;330(Pt 2):737–744. doi: 10.1042/bj3300737. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nichols B. A., Chiappino M. L., Dawson C. R. Demonstration of the mucous layer of the tear film by electron microscopy. Invest Ophthalmol Vis Sci. 1985 Apr;26(4):464–473. [PubMed] [Google Scholar]
- Pemberton L., Taylor-Papadimitriou J., Gendler S. J. Antibodies to the cytoplasmic domain of the MUC1 mucin show conservation throughout mammals. Biochem Biophys Res Commun. 1992 May 29;185(1):167–175. doi: 10.1016/s0006-291x(05)80971-4. [DOI] [PubMed] [Google Scholar]
- Pflugfelder S. C. Tear fluid influence on the ocular surface. Adv Exp Med Biol. 1998;438:611–617. doi: 10.1007/978-1-4615-5359-5_86. [DOI] [PubMed] [Google Scholar]
- Prydal J. I., Artal P., Woon H., Campbell F. W. Study of human precorneal tear film thickness and structure using laser interferometry. Invest Ophthalmol Vis Sci. 1992 May;33(6):2006–2011. [PubMed] [Google Scholar]
- Prydal J. I., Campbell F. W. Study of precorneal tear film thickness and structure by interferometry and confocal microscopy. Invest Ophthalmol Vis Sci. 1992 May;33(6):1996–2005. [PubMed] [Google Scholar]
- Rossi E. A., McNeer R. R., Price-Schiavi S. A., Van den Brande J. M., Komatsu M., Thompson J. F., Carraway C. A., Fregien N. L., Carraway K. L. Sialomucin complex, a heterodimeric glycoprotein complex. Expression as a soluble, secretable form in lactating mammary gland and colon. J Biol Chem. 1996 Dec 27;271(52):33476–33485. doi: 10.1074/jbc.271.52.33476. [DOI] [PubMed] [Google Scholar]
- Sheng Z. Q., Hull S. R., Carraway K. L. Biosynthesis of the cell surface sialomucin complex of ascites 13762 rat mammary adenocarcinoma cells from a high molecular weight precursor. J Biol Chem. 1990 May 25;265(15):8505–8510. [PubMed] [Google Scholar]
- Sheng Z., Wu K., Carraway K. L., Fregien N. Molecular cloning of the transmembrane component of the 13762 mammary adenocarcinoma sialomucin complex. A new member of the epidermal growth factor superfamily. J Biol Chem. 1992 Aug 15;267(23):16341–16346. [PubMed] [Google Scholar]
- Sherblom A. P., Buck R. L., Carraway K. L. Purification of the major sialoglycoproteins of 13762 MAT-B1 and MAT-C1 rat ascites mammary adenocarcinoma cells by density gradient centrifugation in cesium chloride and guanidine hydrochloride. J Biol Chem. 1980 Jan 25;255(2):783–790. [PubMed] [Google Scholar]
- Sherblom A. P., Carraway K. L. A complex of two cell surface glycoproteins from ascites mammary adenocarcinoma cells. J Biol Chem. 1980 Dec 25;255(24):12051–12059. [PubMed] [Google Scholar]
- Sherblom A. P., Huggins J. W., Chesnut R. W., Buck R. L., Ownby C. L., Dermer G. B., Carraway K. L. Cell surface properties of ascites sublines of the 13762 rat mammary adenocarcinoma. Relationship of the major sialoglycoprotein to xenotransplantability. Exp Cell Res. 1980 Apr;126(2):417–426. doi: 10.1016/0014-4827(80)90281-5. [DOI] [PubMed] [Google Scholar]
- Strous G. J., Dekker J. Mucin-type glycoproteins. Crit Rev Biochem Mol Biol. 1992;27(1-2):57–92. doi: 10.3109/10409239209082559. [DOI] [PubMed] [Google Scholar]
- Watanabe H., Fabricant M., Tisdale A. S., Spurr-Michaud S. J., Lindberg K., Gipson I. K. Human corneal and conjunctival epithelia produce a mucin-like glycoprotein for the apical surface. Invest Ophthalmol Vis Sci. 1995 Feb;36(2):337–344. [PubMed] [Google Scholar]
- Williams C. J., Wreschner D. H., Tanaka A., Tsarfaty I., Keydar I., Dion A. S. Multiple protein forms of the human breast tumor-associated epithelial membrane antigen (EMA) are generated by alternative splicing and induced by hormonal stimulation. Biochem Biophys Res Commun. 1990 Aug 16;170(3):1331–1338. doi: 10.1016/0006-291x(90)90540-4. [DOI] [PubMed] [Google Scholar]
- Wu K., Fregien N., Carraway K. L. Molecular cloning and sequencing of the mucin subunit of a heterodimeric, bifunctional cell surface glycoprotein complex of ascites rat mammary adenocarcinoma cells. J Biol Chem. 1994 Apr 22;269(16):11950–11955. [PubMed] [Google Scholar]
