Abstract
The synthesis of the proteins of the apical tight junctions (TJs) depends on a continuous supply of short-chain fatty acids (SCFAs) in colonic epithelium. No studies have evaluated the tissue contents of the TJs proteins in colon segments devoid of a fecal stream. To evaluate the contents of claudin-3 and occludin in the glands of colonic mucosa devoid of a fecal stream. Forty-five rats underwent a diversion of the fecal stream via a left side colostomy and distal mucous fistula. Three groups of 15 animals each were sacrificed at 6, 12 or 18 weeks after surgery. The presence and severity of colitis were defined by histology and inflammation grading scales, respectively. The expression of claudin-3 and occludin were evaluated by immunohistochemistry, and their contents were evaluated by computer-assisted image analysis. Mann–Whitney and Kruskal–Wallis tests were used to evaluate the results at a significance level of 5 % (p < 0.05). The colonic epithelium without a fecal stream had a higher degree of inflammation. Colonic glands without a fecal stream showed a reduction in claudin-3 content independent of the time and reduction in occludin content after 12 weeks of intestinal exclusion. The content of claudin-3 and occludin were mainly reduced at the apical surfaces of the colon glands, whereas segments retaining the fecal stream were maintained. The content of claudin-3 was not reduced with time, although the levels of occludin were reduced after 6 weeks and did not vary thereafter. Deficiencies in SCFAs decreased the content of claudin-3 and occludin in colonic glands with the areas of worst inflammation, confirming the importance of an adequate supply of SCFAs in maintaining the integrity of TJ proteins.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Abdallah DM, Ismael NR (2011) Resveratrol abrogates adhesion molecules and protects against TNBS-induced ulcerative colitis in rats. Can J Physiol Pharmacol 89:811–818. doi:10.1139/Y11-080
Balda MS, Whitney JA, Flores C, Gonzales M, Cereijido M, Matter K (1996) Functional dissociation of paracellular permeability and TER and disruption of the apical–basolateral intramembrane diffusion barrier by expression of a mutant junction membrane protein. J Cell Biol 134:1031–1049
Caltabiano C, Máximo FR, Spadari AP et al (2011) 5-Aminosalicylic acid (5-ASA) can reduce levels of oxidative DNA damage in cells of colonic mucosa with and without fecal stream. Dig Dis Sci 56:1037–1046. doi:10.1007/s10620-010-1378-z
Ciccocioppo R, Finamore A, Ara C, Di Sabatino A, Mengheri E, Corazza GR (2006) Altered expression, localization, and phosphorylation of epithelial junctional proteins in celiac disease. Am J Clin Pathol 125:502–511. doi:10.1309/DTYRA91G8R0KTM8M
Clayburgh DR, Shen L, Turner JR (2004) A porous defense: the leaky epithelial barrier in the intestinal disease. Lab Invest 84:282–291. doi:10.1038/labinvest.3700050
Cunha FL, Silva CMG, Almeida MG et al (2011) Reduction in oxidative stress levels in the colonic mucosa without fecal stream after the application of enemas containing aqueous Ilex paraguariensis extract. Acta Cir Bras 26:289–296. doi:10.1590/S0102-86502011000400008
Dhawan P, Singh AB, Deane NG et al (2005) Claudin-1 regulates cellular transformation and metastatic behavior in colon cancer. J Clin Invest 115:1765–1776. doi:10.1172/JCI24543
Ding L, Lu Z, Lu Q, Chen YH (2013) The claudin family of proteins in human malignancy: a clinical perspective. Cancer Manag Res 5:367–375. doi:10.2147/CMAR.S38294
Edelblum KL, Turner JR (2009) The tight junction in inflammatory disease: communication breakdown. Curr Opin Pharmacol 9:715–720. doi:10.1016/j.coph.2009.06.022
Fries W, Mazzon E, Squarzoni S, Martin A, Martines D, Micali A, Sturniolo GC, Citi S, Longo G (1999) Experimental colitis increases small intestine permeability in the rat. Lab Investig 79:49–57
Furuse M, Hirase T, Itoh M, Nagafuchi A, Yonemura S, Tsukita S (1993) Occludin: a novel integral membrane protein localizing at tight junctions. J Cell Biol 123:1777–1788
Furuse M, Fujita K, Hiiraji T, Fujimoto K, Tsukita S (1998) Claudin-1 and -2: novel integral membrane protein localizing and tight junctions. J Cell Biol 141:1539–1550. doi:10.1083/jcb.141.7.1539
Gassler N, Rohr C, Schneider A et al (2001) Inflammatory bowel disease is associated with changes of enterocytic junctions. Am J Physiol Gastrointest Liver Physiol 281:G216–G228
Gaudier E, Forestier L, Gouyer V, Huet G, Julien R, Hoebler C (2004) Butyrate regulation of glycosylation-related gene expression: evidence for galectin-1 upregulation in human intestinal epithelial goblet cells. Biochem Biophys Res Commun 325:1044–1051. doi:10.1016/j.bbrc.2004.10.141
Gaudier E, Rival M, Buisine MP, Robineau I, Hoebler C (2009) Butyrate enemas upregulate Muc genes expression but decrease adherent mucus thickness in mice colon. Physiol Res 58:111–119
Glotzer DJ, Glick ME, Goldman H (1981) Proctitis following diversion of fecal stream. Gastroenterology 80:438–441
Gupta RB, Harpaz N, Itzkowitz S et al (2007) Histologic inflammation is a risk factor for progression to colorectal neoplasia in ulcerative colitis: a cohort study. Gastroenterology 133:1099–1105. doi:10.1053/j.gastro.2007.08.001
Hirase T, Staddon JM, Saitou M et al (1997) Occludin as a possible determinant of tight junction permeability in endothelial cells. J Cell Sci 110:1603–1613
Hollander D, Vadheim CM, Brettholz E, Petersen GM, Delahunty T, Rotter JI (1986) Increased intestinal permeability in patients with Crohn’s disease and their relatives. A possible etiologic factor. Ann Intern Med 105:883–885. doi:10.7326/0003-4819-105-6-883
John LJ, Fromm M, Schulzke JD (2011) Epithelial barriers in intestinal inflammation. Antioxid Redox Signal 15:1255–1270. doi:10.1089/ars.2011.3892
Kadri CJ, Pereira JA, da Silva CM et al (2013) E-cadherin expression in colonic mucosa with and without fecal stream. J Invest Surg 26:72–79. doi:10.3109/08941939.2012.693334
Kobayashi S, Fujimori T, Mitomi H et al (2014) Immunohistochemical assessment of a unique basal pattern of p53 expression in ulcerative-colitis-associated neoplasia using computer-assisted cytometry. Diagn Pathol 9:99. doi:10.1186/1746-1596-9-99
Kozlowski C, Jeet S, Beyer J et al (2013) An entirely automated method to score DSS-induced colitis in mice by digital image analysis of pathology slides. Dis Model Mech 6:855–865. doi:10.1242/dmm.011759
Kucharzik T, Walsh SV, Chen J, Parkos CA, Nusrat A (2001) Neutrophil transmigration in inflammatory bowel disease is associated with differential expression of epithelial intercellular junction proteins. Am J Pathol 159:2001–2009. doi:10.1016/S0002-9440(10)63051-9
Lal-Nag M, Morin PJ (2009) The claudins. Genome Biol 10:235. doi:10.1186/gb-2009-10-8-235
Lameiro TMM, Silva CMG, Marques LHS et al (2011) Efeitos do butirato nos níveis de peroxidação lipídica em células da mucosa cólica sem trânsito fecal: estudo experimental em ratos. Rev Bras Coloproctol 31:155–164. doi:10.1590/S0101-98802011000200007
Laukoetter MG, Nava P, Nusrat A (2008) Role of the intestinal barrier in inflammatory bowel disease. World J Gastroenterol 14:401–407. doi:10.3748/wjg.14.401
Li Q, Zhang Q, Zhang M, Wang C, Zhu Z, Li N, Li J (2008) Effect of n-3 polyunsaturated fatty acids on membrane microdomain localization of tight junction proteins in experimental colitis. FEBS J 275:411–420. doi:10.1111/j.1742-4658.2007.06210.x
Lu Z, Ding L, Lu Q, Chen YH (2013) Claudins in intestines: distribution and functional significance in health and diseases. Tissue Barriers 1:e24978. doi:10.4161/tisb.24978
Ma X, Fan PX, Li LS, Qiao SY, Zhang GL, Li DF (2012) Butyrate promotes the recovering of intestinal wound healing through its positive effect on the tight junctions. J Anim Sci 90(Suppl 4):266–268. doi:10.2527/jas.50965
Malekian V, Amirfattahi R, Sadri S, Mokhtari M, Aghaie A, Rezaeian M (2013) Computer aided measurement of sub-epithelial collagen band in colon biopsies for collagenous colitis diagnosis. Micron 45:59–67. doi:10.1016/j.micron.2012.10.015
Martinez CAR, Ribeiro ML, Gambero A, Miranda DDC, Pereira JA, Nadal SR (2010a) The importance of oxygen free radicals in the etiopathogenesis of diversion colitis in rats. Acta Cir Bras 25:387–395. doi:10.1590/S0102-86502010000500002
Martinez CAR, Nonose R, Spadari APP et al (2010b) Quantification by computerized morphometry of tissue levels of sulfomucins and sialomucins in diversion colitis in rats. Acta Cir Bras 25:231–240. doi:10.1590/S0102-86502010000300004
Martinez CAR, Fabris FM, Silva CMG, Rodrigues MR, Sato DT, Pereira JA (2012) Oxidative stress and changes in the content and pattern of tissue expression of β-catenin protein in diversion colitis. J Coloproctol (Rio J) 32:343–358. doi:10.1590/S2237-93632012000400001
Martinez CA, de Almeida MG, da Silva CM et al (2013) Enemas with N-acetylcysteine can reduce the level of oxidative damage in cells of the colonic mucosa diverted from the faecal stream. Dig Dis Sci 58(12):3452–3459. doi:10.1007/s10620-013-2768-9
Mees ST, Mennigen R, Spieker T et al (2009) Expression of tight and adherens junctions proteins in ulcerative colitis associated colorectal carcinomas: upregulation of claudin-1, claudin-3, claudin-4, and β-catenin. Int J Colorectal Dis 24:361–368. doi:10.1007/s00384-009-0653-y
Mitic LL, Anderson JM (1998) Molecular architecture of tight junctions. Annu Rev Physiol 60:121–142. doi:10.1146/annurev.physiol.60.1.121
Nassri CGG, Nassri AB, Favero E, Rotta CM, Martinez CAR, Margarido NF (2008) Influência da irrigação de soluções nutricionais no colo excluso de trânsito intestinal: estudo experimental em ratos. Rev Bras Coloproctol 28:306–314. doi:10.1590/S0101-98802008000300006
Nazli A, Wang A, Steen O et al (2006) Enterocyte cytoskeleton changes are crucial for enhanced translocation of nonpathogenic Escherichia coli across metabolically stressed gut epithelia. Infect Immun 74:192–201. doi:10.1128/IAI.74.1.192-201.2006
Nonose R, Spadari APP, Priolli DG, Máximo FR, Pereira JA, Martinez CAR (2009) Tissue quantification of neutral and acid mucins in the mucosa of the colon with and without fecal stream in rats. Acta Cir Bras 24:267–275. doi:10.1590/S0102-86502009000400005
Ozawa M, Ringwald M, Kemler R (1990) Uvomorulin–catenin complex formation is regulated by a specific domain in the cytoplasmic region of the cell adhesion molecule. Proc Natl Acad Sci USA 87:4246–4250
Pacheco RG, Esposito CC, Müller LC et al (2012) Use of butyrate or glutamine in enema solution reduces inflammation and fibrosis in experimental diversion colitis. World J Gastroenterol 28(18):4278–4287. doi:10.3748/wjg.v18.i32.4278
Peng L, Li ZR, Green RS, Holzman IR, Lin J (2009) Butyrate enhances the intestinal barrier by facilitating tight junction assembly via activation of AMP-activated protein kinase in Caco-2 cell monolayers. J Nutr 139:1619–1625. doi:10.3945/jn.109.104638
Pereira JA, Rodrigues MR, Sato DT et al (2013) Evaluation of sucralfate enema in experimental diversion colitis. J Coloproctol (Rio J) 33:182–190. doi:10.1016/j.jcol.2013.08.005
Plöger S, Stumpff F, Penner GB et al (2012) Microbial butyrate and its role for barrier function in the gastrointestinal tract. Ann NY Acad Sci 1258:52–59. doi:10.1111/j.1749-6632.2012.06553.x
Prasad S, Mingrino R, Kaukinen K et al (2005) Inflammatory processes have differential effects on claudins 2, 3 and 4 in colonic epithelial cells. Lab Invest 85:1139–1162. doi:10.1038/labinvest.3700316
Pravda J (2005) Radical induction theory of ulcerative colitis. World J Gastroenterol 11:2371–2384
Resnick MB, Konkin T, Routhier J (2005) Claudin-1 is a strong prognostic indicator in stage II colonic cancer: a tissue microarray study. Mod Pathol 18:511–518. doi:10.1038/modpathol.3800301
Saitou M, Furuse M, Sasaki H et al (2000) Complex phenotype of mice lacking occludin, a component of tight junction strands. Mol Biol Cell 11:4131–4142
Sauer J, Richter KK, Pool-Zobel BL (2007) Physiological concentrations of butyrate favorably modulate genes of oxidative and metabolic stress in primary human colon cells. J Nutr Biochem 18:736–745
Schmitz H, Barmeyer C, Fromm M et al (1999) Altered tight junction structure contributes to the impaired epithelial barrier function in ulcerative colitis. Gastroenterology 116:301–309
Schneeberger EE, Lynch RD (1984) Tight junctions. Their structure, composition, and function. Circ Res 55:723–733
Sousa MV, Priolli DG, Portes AV, Cardinalli IA, Pereira JA, Martinez CAR (2008) Evaluation by computerized morphometry of histopathological alterations of the colon wall in segments with and without intestinal transit in rats. Acta Cir Bras 23:417–424. doi:10.1590/S0102-86502008000500005
Su L, Shen L, Clayburgh DR et al (2009) Targeted epithelial tight junction dysfunction causes immune activation and contributes to development of experimental colitis. Gastroenterology 136:551–563. doi:10.1053/j.gastro.2008.10.081
Turner JR (2009) Intestinal mucosal barrier function in health and disease. Nat Rev Immunol 9:799–809. doi:10.1038/nri2653
Usami Y, Chiba H, Nakayama F et al (2006) Reduced expression of claudin-7 correlates with invasion and metastasis in squamous cell carcinoma of the esophagus. Hum Pathol 37:569–577. doi:10.1016/j.humpath.2005.12.018
Van Itallie CM, Anderson JM (2014) Architecture of tight junctions and principles of molecular composition. Semin Cell Dev Biol 36C:157–165. doi:10.1016/j.semcdb.2014.08.011
Van Itallie CM, Mitic LL, Anderson JM (2000) Claudin-2 forms homodimers and is a component of a high molecular weight protein complex. J Biol Chem 286:3442–3450. doi:10.1074/jbc.M110.195578
Van Itallie CM, Holmes J, Bridges A, Anderson JM (2009) Claudin-2-dependent changes in noncharged solute flux are mediated by the extracellular domains and require attachment to the PDZ-scaffold. Ann NY Acad Sci 1165:82–87. doi:10.1111/j.1749-6632.2009.04052.x
Vieira EL, Leonel AJ, Sad AP et al (2012) Oral administration of sodium butyrate attenuates inflammation and mucosal lesion in experimental acute ulcerative colitis. J Nutr Biochem 23:430–436. doi:10.1016/j.jnutbio.2011.01.007
Watson AJ, Chu S, Sieck L, Gerasimenko O, Bullen T, Campbell F, Ckenna M, Rose T, Montrose MH (2005) Epithelial barrier function in vivo sustained despite gaps in epithelial layers. Gastroenterol 129:902–912
Weber CR, Nalle SC, Tretiakova M, Rubin DT, Turner JR (2008) Claudin-1 and claudin-2 expression is elevated in inflammatory bowel disease and may contribuite to early neoplastic transformation. Lab Invest 88:1110–1120. doi:10.1038/labinvest.2008.78
Wongdee K, Teerapornpuntakit J, Siangpro C, Chaipai S, Charoenphandhu N (2013) Duodenal villous hypertrophy and upregulation of claudin-15 protein expression in lactating rats. J Mol Histol 44:103–109
Wu SJ, Don TM, Lin CW, Mi FL (2014) Delivery of berberine using chitosan/fucoidan–taurine conjugate nanoparticles for treatment of defective intestinal epithelial tight junction barrier. Mar Drugs 12:5677–5697. doi:10.3390/md12115677
Zeissig S, Burgel N, Gunzel D et al (2007) Changes in expression and distribution of claudin 2, 5 and 8 lead to discontinuous tight junctions and barrier dysfunction in active Crohn’s disease. Gut 56:61–72
Acknowledgments
Fundação de Amparo a Pesquisa do Estado de São Paulo—FAPESP: Project No. 2010/12.492-7.
Conflict of interest
The authors declare that they have no conflict of interest.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Martinez, C.A.R., de Campos, F.G.C.M., de Carvalho, V.R.J. et al. Claudin-3 and occludin tissue content in the glands of colonic mucosa with and without a fecal stream. J Mol Hist 46, 183–194 (2015). https://doi.org/10.1007/s10735-015-9610-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10735-015-9610-y