Abstract
Tight junctions are essential for the maintenance of epithelial cell polarity. We have previously established a system for the primary culture of salivary parotid acinar cells that retain their ability to generate new secretory granules and to secrete proteins in a signal-dependent manner. Because cell polarity and cell-cell adhesion are prerequisites for the formation of epithelial tissues, we have investigated the structure of the tight junctions in these cultures. We have found two types of cellular organization in the culture: monolayers and semi-spherical clusters. Electron microscopy has revealed tight junctions near the apical region of the lateral membranes between cells in the monolayers and cells at the surface of the clusters. The cells in the interior of the clusters also have tight junctions and are organized around a central lumen. These interior cells retain more secretory granules than the surface or monolayer cells, suggesting that they maintain their original character as acinar cells. The synthesis of claudin-4 increases during culture, although it is not detectable in the cells immediately after isolation from the glands. Immunofluorescence microscopy has shown that claudin-4 is synthesized in the monolayers and at the surface of the clusters, but not inside the clusters. Only claudin-3, which is present in the original acinar cells following isolation and in the intact gland, has been detected inside the clusters. These results suggest that differences in claudin expression are related to the three-dimensional structures of the cell cultures and reflect their ability to function as acinar cells.
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References
Bilder D (2004) Epithelial polarity and proliferation control: links from the Drosophila neoplastic tumor suppressors. Genes Dev 18:1909–1925
Blanchard AA, Watson PH, Shiu RP, Leygue E, Nistor A, Wong P, Myal Y (2006) Differential expression of claudin 1, 3, and 4 during normal mammary gland development in the mouse. DNA Cell Biol 25:79–86
Brizuela BJ, Wessely O, De Robertis EM (2001) Overexpression of the Xenopus tight-junction protein claudin causes randomization of the left-right body axis. Dev Biol 230:217–229
Chen SP, Zhou B, Willis BC, Sandoval AJ, Liebler JM, Kim KJ, Ann DK, Crandall ED, Borok Z (2005) Effects of transdifferentiation and EGF on claudin isoform expression in alveolar epithelial cells. J Appl Physiol 98:322–328
Daugherty BL, Mateescu M, Patel AS, Wade K, Kimura S, Gonzales LW, Guttentag S, Ballard PL, Koval M (2004) Developmental regulation of claudin localization by fetal alveolar epithelial cells. Am J Physiol 287:L1266–L1273
Fujita-Yoshigaki J, Dohke Y, Hara-Yokoyama M, Kamata Y, Kozaki S, Furuyama S, Sugiya H (1996) Vesicle-associated membrane protein 2 is essential for cAMP-regulated exocytosis in rat parotid acinar cells: the inhibition of cAMP-dependent amylase release by botulinum neurotoxin B. J Biol Chem 271:13130–13134
Fujita-Yoshigaki J, Tagashira A, Yoshigaki T, Furuyama S, Sugiya H (2005) A primary culture of parotid acinar cells retaining capacity for agonists-induced amylase secretion and generation of new secretory granules. Cell Tissue Res 320:455–464
Furuse M, Tsukita S (2006) Claudins in occluding junctions of humans and flies. Trends Cell Biol 16:181–188
Furuse M, Sasaki H, Tsukita S (1999) Manner of interaction of heterogeneous claudin species within and between tight junction strands. J Cell Biol 147:891–903
Furuse M, Furuse K, Sasaki H, Tsukita S (2001) Conversion of zonulae occludentes from tight to leaky strand type by introducing claudin-2 into Madin-Darby canine kidney I cells. J Cell Biol 153:263–272
Gonzalez-Mariscal L, Betanzos A, Nava P, Jaramillo BE (2003) Tight junction proteins. Prog Biophys Mol Biol 81:1–44
Haouzi D, Baghdiguian S, Granier G, Travo P, Mangeat P, Hibner U (2005) Three-dimensional polarization sensitizes hepatocytes to Fas/CD95 apoptotic signalling. J Cell Sci 118:2763–2773
Hashizume A, Ueno T, Furuse M, Tsukita S, Nakanishi Y, Hieda Y (2004) Expression patterns of claudin family of tight junction membrane proteins in developing mouse submandibular gland. Dev Dyn 231:425–431
Heiskala M, Peterson PA, Yang Y (2001) The roles of claudin superfamily proteins in paracellular transport. Traffic 2:93–98
Hoffman MP, Kibbey MC, Letterio JJ, Kleinman HK (1996) Role of laminin-1 and TGF-beta 3 in acinar differentiation of a human submandibular gland cell line (HSG). J Cell Sci 109:2013–2021
Kibbey MC, Royce LS, Dym M, Baum BJ, Kleinman HK (1992) Glandular-like morphogenesis of the human submandibular tumor cell line A253 on basement membrane components. Exp Cell Res 198:343–351
Kobayashi K, Utsumi H, Okada M, Sakairi T, Ikeda I, Kusakabe M, Takagi S (2003) One-step RT-PCR without initial RNA isolation step for laser-microdissected tissue sample. J Vet Med Sci 65:917–919
Kojima S, Rahner C, Peng S, Rizzolo LJ (2002) Claudin 5 is transiently expressed during the development of the retinal pigment epithelium. J Membr Biol 186:81–88
Lam K, Zhang L, Bewick M, Lafrenie RM (2004) HSG cells differentiated by culture on extracellular matrix involves induction of S-adenosylmethione decarboxylase and ornithine decarboxylase. J Cell Physiol 203:353–361
Lui WY, Lee WM, Cheng CY (2001) Transforming growth factor-beta3 perturbs the inter-Sertoli tight junction permeability barrier in vitro possibly mediated via its effects on occludin, zonula occludens-1, and claudin-11. Endocrinology 142:1865–1877
Matter K, Aijaz S, Tsapara A, Balda MS (2005) Mammalian tight junctions in the regulation of epithelial differentiation and proliferation. Curr Opin Cell Biol 17:453–458
Morita K, Furuse M, Fujimoto K, Tsukita S (1999) Claudin multigene family encoding four-transmembrane domain protein components of tight junction strands. Proc Natl Acad Sci USA 96:511–516
Ojakian GK, Ratcliffe DR, Schwimmer R (2001) Integrin regulation of cell-cell adhesion during epithelial tubule formation. J Cell Sci 114:941–952
Peppi M, Ghabriel MN (2004) Tissue-specific expression of the tight junction proteins claudins and occludin in the rat salivary glands. J Anat 205:257–266
Saitou M, Fujimoto K, Doi Y, Itoh M, Fujimoto T, Furuse M, Takano H, Noda T, Tsukita S (1998) Occludin-deficient embryonic stem cells can differentiate into polarized epithelial cells bearing tight junctions. J Cell Biol 141:397–408
Schneeberger EE, Lynch RD (1992) Structure, function, and regulation of cellular tight junctions. Am J Physiol 262:L647–L661
Schneeberger EE, Lynch RD (2004) The tight junction: a multifunctional complex. Am J Physiol 286:C1213–C1228
Tsukita S, Furuse M (2002) Claudin-based barrier in simple and stratified cellular sheets. Curr Opin Cell Biol 14:531–536
Tsukita S, Furuse M, Itoh M (2001) Multifunctional strands in tight junctions. Nat Rev Mol Cell Biol 2:285–293
Turksen K, Troy TC (2004) Barriers built on claudins. J Cell Sci 117:2435–2447
Umeda K, Ikenouchi J, Katahira-Tayama S, Furuse K, Sasaki H, Nakayama M, Matsui T, Tsukita S, Furuse M, Tsukita S (2006) ZO-1 and ZO-2 independently determine where claudins are polymerized in tight-junction strand formation. Cell 126:741–754
Van Itallie CM, Anderson JM (2004) The role of claudins in determining paracellular charge selectivity. Proc Am Thorac Soc 1:38–41
Van Itallie C, Rahner C, Anderson JM (2001) Regulated expression of claudin-4 decreases paracellular conductance through a selective decrease in sodium permeability. J Clin Invest 107:1319–1327
Zahraoui A, Louvard D, Galli T (2000) Tight junction, a platform for trafficking and signaling protein complexes. J Cell Biol 151:F31–F36
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This work was supported by grants-in-aid for scientific research from the Ministry of Education, Science, Culture, Sports, and Technology of Japan (16591868, 16791135), by a Suzuki Memorial Grant of the Nihon University School of Dentistry at Matsudo (Joint Research Grant for 2003), by a Nihon University Multidisciplinary Research Grant for 2005 and 2006, and by a Grant-in-Aid for a 2003 Multidisciplinary Research Project from MEXT.
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Qi, B., Fujita-Yoshigaki, J., Michikawa, H. et al. Differences in claudin synthesis in primary cultures of acinar cells from rat salivary gland are correlated with the specific three-dimensional organization of the cells. Cell Tissue Res 329, 59–70 (2007). https://doi.org/10.1007/s00441-007-0389-3
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DOI: https://doi.org/10.1007/s00441-007-0389-3