Abstract
Progress in understanding how mucosal surfaces are protected is closely related to the development of morphologic techniques to study the structure and secretory function of the mucosal epithelia. Morphologic methods have allowed characterization of mucus-secreting cells of the epithelia of the eye, and the respiratory, gastrointestinal (GI), and reproductive tracts. Characteristics of the mucus-secreting cells of these tissues vary, and many questions remain regarding special characteristics of mucus present over the differing mucosal surfaces. Recent progress in cloning and characterization of mucin genes has facilitated the use of in situ hybridization (ISH) to begin to characterize the mucin gene repertoires and specific functions of mucins expressed by the various epithelia, either those covering mucosal surfaces or glandular epithelia contributing to the mucous layer on the surface of the tissue. ISH has been a particularly valuable method in this regard, since antibodies to specific mucin proteins are often difficult to use on tissues or secretions without heroic methods to deglycosylate in order to make protein epitopes available.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Carlstedt I., Sheehan J. K., Corfield A. P., and Gallagher J. T. (1995) Mucous glycoproteins: A gel of a problem. Essay Biochem. 20, 40–76.
Strous G. J. and Dekker J. (1992) Mucin-type glycoproteins. Crit. Rev. Biochem. Mol. Biol. 27, 57–92.
Gum J. R.,Jr. (1992) Mucin genes and the proteins they encode: Structure diversity and regulation. Am. J. Respir. Cell. Mol. Biol. 7, 557–564.
Gendler S. J. and Spicer A. P. (1995) Epithelial mucin genes. Annu. Rev. Physiol. 57, 607–634.
Gum J. R.,Jr. (1995) Mucins: Their structure and biology. Biochem. Soc. Trans. 23, 795–799.
Choo K. H. A., ed. (1994) In situ Hybridization Protocols, Humana, Inc., Totowa, NJ.
Inatomi T., Spurr-Michaud S., Tisdale A. S., and Gipson I. K. (1995) Human corneal and conjunctival epithelia express MUC1 mucin. Invest. Ophthalmol. Vis. Sci. 36, 1818–1827.
Inatomi T., Spurr-Michaud S., Tisdale A. S., Zhan Q., Feldman S. T., and Gipson I. K. (1996) Expression of secretory mucin genes by human conjunctival epithelia. Invest. Ophthalmol. Vis. Sci. 37, 1684–1692.
Gipson I. K. and Inatomi T. (1999) Cellular origin of mucins of the ocular surface. in Advances in Experimental Medicine and Biology (Sullivan D. A., ed.), Plenum, New York, pp. 221–228.
Gipson I. K., Ho S. B., Spurr-Michaud S. J., Tisdale A. S., Zhan Q., Torlakovic E., Pudney J., Anderson D. J., Toribara N. W., and Hill J. A.,III. (1997) Mucin genes expressed by human female reproductive tract epithelia. Biol. Reprod. 56, 999–1011.
Shekels L. L., Hunninghake D. A., Tisdale A. S., Gipson I. K., Kieliszewski M., Kozak C. A., and Ho S. B. (1998) Cloning and characterization of mouse intestinal MUC3 mucin: 3′ sequence contains EGF-like domains. Biochem. J. 300, 1301–1308.
Polak J. M. and McGee J. O. D., eds. (1990) In Situ Hybridization: Principles and Practice. Oxford University Press, New York.
Young W. S.,III (1990) In situ hybridization histochemistry, in Handbook of Chemical Neuroanatomy, (Björklund A., Hökfelt R., Wouierlood F. G., and van den Pol A. N., eds.), Elsevier Science Publishers, UK, pp. 481–512.
Ausubel F. M., Brent R., Kingston R. E., Moore D. D., Seidman J. G., Smith J. A., and Struhl K. (1995) In situ hybridization and immunohistochemistry. Curr. Protocols Mol. Biol. 2, 14.0.1–14.8.24.
Millen K. J. and Hui C.-C. (1996) Radioactive in situ hybridization of tissue sections, in A Laboratory Guide to RNA: Isolation, Analysis, and Synthesis (Krieg P. A., ed.), Wiley-Liss, New York, pp. 339–355.
Penschow J. D. and Coghlan J. P. (1994) Hybridization histochemistry using radiolabeled oligodeoxyribonucleotide probes, in In Situ Hybridization Protocols, (Andy Choo K. H., ed.), Humana, Totowa, NJ, pp. 277–292.
Beck F., Tucci J., and Senior P. V. (1994) Tissue distribution of gene expression in mammalian development, in In Situ Hybridization Protocols (Choo K. H. A., ed.), Humana, Totowa, NJ, pp. 313–344.
Gait M. J., ed. (1984) Oligonucleotide Synthesis: A Practical Approach, IRL, Oxford, UK.
Cowell I. G. (1997) Cloning sequence-specific DNA-binding factors from cDNA expression libraries using oligonucleotide binding site probes, in cDNA Library Protocols, (Cowell I. G. and Austin C. A., eds.), Humana, Totowa, NJ, pp. 161–170.
Chambers J. A., Hollingsworth M. A., Trezise A. E. O., and Harris A. (1994) Developmental expression of mucin genes MUC1 and MUC2. J. Cell. Sci. 107, 413–424.
Balagué C., Audié J.-P., Porchet N., and Real F. X. (1995) In situ hybridization shows distinct patterns of mucin gene expression in normal, benign, and malignant pancreas tissues. Gastroenterology 109, 953–964.
Jany B. H., Gallup M. W., Yan P.-S., Gum J. R., Kim Y. S., and Basbaum C. B. (1991) Human bronchus and intestine express the same mucin gene. J. Clin. Invest. 88, 77–82.
Li D., Wang D., Majumdar S., Jany B., Durham S. R., Cottrell J., Caplen N., Geddes D. M., Alton E. W., and Jeffery P. K. (1997) Localization and up-regulation of mucin (MUC2) gene expression in human nasal biopsies of patients with cystic fibrosis. J. Pathol. 181, 305–310.
Audie J. P., Janin A., Porchet N., Copin M. C., Gosselin B., and Aubert J. P. (1993) Expression of human mucin genes in respiratory, digestive, and reproductive tracts ascertained by in situ hybridization. J. Histochem. Cytochem. 41, 1479–1485.
Byrd J. C., Yan P., Sternberg L., Yunker C. K., Scheiman J. M., and Bresalier R. S. (1997) Aberrant expression of gland-type gastric mucin in the surface epithelium of Helicobacter pylori-infected patients. Gastroenterology 113, 455–464.
Keates A. C., Nunes D. P., Afdhal N. H., Troxler R. F., and Offner G. D. (1997) Molecular cloning of a major human gall bladder mucin: complete C-terminal sequence and genomic organization of MUC5B. Biochem. J. 324, 295–303.
Nielsen P. A., Bennett E. P., Wandall H. H., Therkildsen M. H., Hannibal J., and Clausen H. (1997) Identification of a major human high molecular weight salivary mucin (MG1) as tracheobronchial mucin MUC5B. Glycobiology 7, 413–419.
Reid C. J., Hyde K., Ho S. B., and Harris A. (1997) Cystic fibrosis of the pancreas: Involvement of MUC6 mucin in obstruction of pancreatic ducts. Mol. Med. 3, 403–411.
Bolos C. D., Garrido M., and Real F. X. (1995) MUC6 apomucin shows a distinct normal tissue distribution that corrrelates with Lewis antigen expression in the human stomach. Gastroenterology 109, 723–734.
Aust M. R., Madsen C. S., Jennings A., Kasperbauer J. L., and Gendler S. J. (1997) Mucin mRNA expression in normal and vasomotor inferior turbinates. Am. J. Rhinol. 11, 293–302.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2000 Humana Press Inc.
About this protocol
Cite this protocol
Gipson, I.K. (2000). In Situ Hybridization Techniques for Localizing Mucin mRNA. In: Corfield, A.P. (eds) Glycoprotein Methods and Protocols. Methods in Molecular Biology™, vol 125. Humana Press. https://doi.org/10.1385/1-59259-048-9:323
Download citation
DOI: https://doi.org/10.1385/1-59259-048-9:323
Publisher Name: Humana Press
Print ISBN: 978-0-89603-720-5
Online ISBN: 978-1-59259-048-3
eBook Packages: Springer Protocols