The Journal of Membrane Biology

, Volume 202, Issue 1, pp 39–49 | Cite as

Lanthanum Effect on the Dynamics of Tight Junction Opening and Closing

  • F. Lacaz-Vieira
  • M. M. Marques


We present a comparative study in frog urinary bladders (FUB) and A6 cell monolayers (A6CM) on the effect of La3+ on tight junction (TJ) dynamics. These tissues react similarly to changes of basolateral Ca2+ (Ca2+ bl), while responding differently to the action of La3+ bl. In FUB, La3+ bl shows a Ca2+-antagonistic effect that promotes TJ opening in the presence of a normal Ca2+ bl concentration. In A6CM, in contrast, La3+ bl always shows a clear Ca2+-agonistic effect. The fact that a concentration of La3+ bl one fifth of the normal Ca2+ bl leads in FUB to TJ opening and in A6CM to a complete recovery of the TJ seal indicates a high affinity of La3+ for the Ca2+-binding sites in both tissues. In FUB, apical La3+ (La3+ ap) exhibits, differently from its basolateral effect, an evident Ca2+-agonistic effect, suggesting a dual effect of La3+, depending on which side of the bladder La3+ is applied. In A6CM La3+ ap has a Ca2+-agonistic effect similar to La3+ bl. The effects of La3+ bl in FUB and in A6CM are consistent, according to our previous publications, with La3+ acting antagonistically or agonistically, respectively, on the Ca2+ binding sites of zonula adhaerens. Despite the fact that the effect of La3+ ap is clear in both tissues, its site of action is yet to be determined. Protonation of the Ca2+-binding sites causes a decrease of its agonistic effect on A6CM, consistent with a negatively charged binding site. In A6CM La3+ apparently replaces Ca2+, mimicking the effect of Ca2+ triggering the cascade of events leading to TJ closure. In FUB, La3+ interacts with the binding sites, dislodging Ca2+, with a high affinity, but this interaction is inadequate to initiate or sustain the process of junction closing. Possibly, the difference between the two preparations resides in subtle conformation differences of the outer segment of E-cadherin molecules.


Calcium Lanthanum Tight junction E-cadherin A6 cells Frog urinary bladders 



Supported by grants from Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP-99/03077-5) and Conselho Nacional de Desenvolvimento Científíco e Tecnológico (CNPq-300500/2003-3).


  1. Alvarado, R.H., Dietz, T.H., Mullen, T.L. 1975Chloride transport across isolated skin of Rana pipiens Am. J. Physiol.229896876PubMedGoogle Scholar
  2. Anderson, J.M., Itallie, C.M. 1999Tight junctions: closing in on the sealCurr. Biol.9R922R924CrossRefPubMedGoogle Scholar
  3. Armstrong, C.M., Cota, G. 1990Modification of sodium channel gating by lanthanum. Some effects that cannot be explained by surface charge theoryJ. Gen. Physiol.9611291140CrossRefPubMedGoogle Scholar
  4. Balda, M.S., Fallon, M.B., Itallie, C.M., Anderson, J.M. 1992Structure, regulation, and pathophysiology of tight junctions in the gastrointestinal tractYale J. Biol. Med.65725735PubMedGoogle Scholar
  5. Balda, M.S., Gonzalez-Mariscal, L., Contreras, R.G., Macias-Silva, M., Torres-Marquez, M.E., García Sáinz, J.A., Cereijido, M. 1991Assembly and sealing of tight junctions: Possible participation of G-proteins, phospholipase C, protein kinase C and calmodulinJ. Membrane Biol.122193202Google Scholar
  6. Balda, M.S., González-Mariscal, L., Matter, K., Cereijido, M., Anderson, J.M. 1993Assembly of the tight junction: The role of diacylglycerolJ. Cell Biol.123293302CrossRefPubMedGoogle Scholar
  7. Boller, K., Vestweber, D., Kemler, R. 1985Cell-adhesion molecule uvomorulin is localized in the intermediate junction of adult intestinal epithelial cellsJ. Cell. Biol.100327332CrossRefPubMedGoogle Scholar
  8. Brown, R.C., Davis, T.P. 2002Calcium modulation of adherens and tight junction function: a potential mechanism for blood-brain barrier disruption after strokeStroke3317061711CrossRefPubMedGoogle Scholar
  9. Castro, J.A., Sesso, A., Lacaz-Vieira, F. 1993Deposition of BaSO4 in the tight junctions of amphibian epithelia causes their opening; apical Ca2+ reverses this effectJ. Membrane Biol.1341529CrossRefGoogle Scholar
  10. Cereijido, M., Ehrenfeld, J., Meza, I., Martinez-Palomo, A. 1980Structural and functional membrane polarity in cultured monolayers of MDCK cellsJ. Membrane Biol.52147159Google Scholar
  11. Cereijido, M., González-Mariscal, L., Avila, G., Contreras, R.G. 1988Tight junctionsCRC Crit. Rev. Anat. Sci.1171192Google Scholar
  12. Cereijido, M., Meza, I., Martinez-Palomo, A. 1981Occluding junctions in cultured epithelial monolayersAm. J. Physiol.240C96C102PubMedGoogle Scholar
  13. Cereijido, M., Ponce, A., González-Mariscal, L. 1989Tight junctions and apical/basolateral polarityJ. Membrane Biol.11019Google Scholar
  14. Cereijido, M., Valdés, J., Shoshani, L., Contreras, R.G. 1998Role of tight junctions in establishing and maintaining cell polarityAnnu. Rev. Physiol.60161177CrossRefPubMedGoogle Scholar
  15. Clarke, B.L., Moore, D.R., Blalock, J.E. 1994Adrenocorticotropic hormone stimulates a transient calcium uptake in rat lymphocytesEndocrinology13517801786CrossRefPubMedGoogle Scholar
  16. Galli, P., Camilli, B.P., Meldolesi, J. 1976Extracellular calcium and the organization of tight junctions in pancreatic acinar cellsExp. Cell Res.99178183CrossRefPubMedGoogle Scholar
  17. García-Colunga, J., Miledi, R. 1997Opposite effects of lanthanum on different types of nicotinic acetylcholine receptorsNeuroReport832933296PubMedGoogle Scholar
  18. Gleeson, R.A., McDowell, L.M., Aldrich, H.C., Hammar, K., Smith, P.J. 2000Sustaining olfaction at low salinities: evidence for a paracellular route of ion movement from the hemolymph to the sensillar lymph in the olfactory sensilla of the blue crab Callinectes sapidusCell Tissue Res.301423431CrossRefPubMedGoogle Scholar
  19. Gonzalez-Mariscal, L., Betanzos, A., Nava, P., Jaramillo, B.E. 2003Tight junction proteinsProg. Biophys. Mol. Biol.81144CrossRefPubMedGoogle Scholar
  20. González-Mariscal, L., ChávezRamirez, B., Cereijido, M. 1985Tight junction formation in cultured epithelial cells (MDCK)J. Membrane Biol.86113125Google Scholar
  21. Gottardi, C.J., Wong, E., Gumbiner, B.M. 2001E-cadherin suppresses cellular transformation by inhibiting beta-catenin signaling in an adhesion-independent mannerJ. Cell Biol.15310491060CrossRefPubMedGoogle Scholar
  22. Gumbiner, B., Stevenson, B.R., Grimaldi, A. 1988The role of the cell adhesion molecule uvomorulin in the formation and maintenance of epithelial junctional complexJ. Cell Biol.10715751587CrossRefPubMedGoogle Scholar
  23. Hays, R.M., Singer, B., Malamed, S. 1965The effect of calcium withdrawal on the structure and function of the toad bladderJ. Cell. Biol.25195208CrossRefPubMedGoogle Scholar
  24. Hille, B., Woodhull, A.M., Shapiro, B.I. 1975Negative surface charge near sodium channels of nerve: divalent ions, monovalent ions and pHPhilos. Trans. R. Soc. Lond. B.270301318Google Scholar
  25. Jan, C.R., Ho, C.M., Wu, S.N., Tseng, C.J. 1999Mechanism of rise and decay of thapsigargin-evoked calcium signals in MDCK cellsLife Sci.64259267CrossRefPubMedGoogle Scholar
  26. Kassab, F.,Jr., Marques, R.P., Lacaz-Vieira, F. 2002Modeling tight junction dynamics and oscillationsJ. Gen. Physiol.120237247CrossRefPubMedGoogle Scholar
  27. Kemler, R. 1992Classical cadherinsSemin. Cell Biol.3149155PubMedGoogle Scholar
  28. Koch, A.W., Pokutta, S., Lustig, A., Engel, J. 1997Calcium binding and homoassociation of E-cadherin domainsBiochemistry3676977705CrossRefPubMedGoogle Scholar
  29. Lacaz-Vieira, F. 1986Sodium flux in the apical membrane of the toad skin: aspects of its regulation and the importance of the ionic strength of the outer solution upon the reversibility of amiloride inhibitionJ. Membrane Biol.922736Google Scholar
  30. Lacaz-Vieira, F. 1997Calcium site specificity - Early Ca2+-related tight junction eventsJ. G. Physiol.110727740CrossRefGoogle Scholar
  31. Lacaz-Vieira, F. 2000Tight junction dynamics: oscillations and the role of protein kinase CJ. Membrane Biol.178151161CrossRefGoogle Scholar
  32. Lacaz-Vieira, F., Jaeger, M.M. 2001Protein kinase inhibitors and the dynamics of tight junction opening and closing in A6 cell monolayersJ. Membrane Biol.184185196CrossRefGoogle Scholar
  33. Lacaz-Vieira, F., Kachar, B. 1996Tight junction dynamics in the frog urinary bladderCell Adhes. Commun.45368PubMedGoogle Scholar
  34. Lacaz-Vieira, F., Jaeger, M., Farshori, P., Kachar, B. 1999Small synthetic peptides homologous to segments of the first external loop of occludin impair tight junction resealingJ. Membrane Biol.168289297CrossRefGoogle Scholar
  35. Lippoldt, A., Kniesel, U., Liebner, S., Kalbacher, H., Kirsch, T., Wolburg, H., Haller, H. 2000Structural alterations of tight junctions are associated with loss of polarity in stroke-prone spontaneously hypertensive rat blood-brain barrier endothelial cellsBrain Res.885251261CrossRefPubMedGoogle Scholar
  36. Lora, L., Mazzon, E., Martines, D., Fries, W., Muraca, M., Martin, A., d’Odorico, A., Naccarato, R., Citi, S. 1997Hepatocyte tight-junctional permeability is increased in rat experimental colitisGastroenterology11313471354PubMedGoogle Scholar
  37. Machen, T.E., Erlij, D., Wooding, F.B.P. 1972Permeability junctional complexes. The movement of lanthanum across rabbit gallbladder and intestineJ. Cell Biol.54302312CrossRefPubMedGoogle Scholar
  38. Martinez-Palomo, A., Meza, I., Beaty, G., Cereijido, M. 1980Experimental modulation of occluding junctions in a cultured transporting monolayerJ. Cell Biol.87736745CrossRefPubMedGoogle Scholar
  39. Mazzon, E., Sturniolo, G.C., Puzzolo, D., Frisina, N., Fries, W. 2002Effect of stress on the paracellular barrier in the rat ileumGut51507513CrossRefPubMedGoogle Scholar
  40. Meldolesi, J., Castiglioni, G., Parma, R., Nassivera, N., Camilli, P. 1978Ca2+-dependent disassembly and reassembly of occluding junctions in guinea pig pancreatic acinar cellsJ. Cell Biol.79156172CrossRefPubMedGoogle Scholar
  41. Nagar, B., Overduin, M., Ikura, M., Rini, J.M. 1996Structural basis of calcium-induced E-cadherin rigidification and dimerizationNature380360364CrossRefPubMedGoogle Scholar
  42. Nelson, M.T. 1987Effects of permeant ions and blockers on properties of single calcium channels from brainsHille, B.Frambrough, D.M. eds. Proteins of Excitable MembranesSoc. Gen. Physiol. and Wiley-lnterscienceNew York215230Google Scholar
  43. Neter, J., Wasserman, W. 1974Applied Linear Statistical Models: Regression Analysis of Variance and Experimental DesignsRichard D. Irwin, Inc.Homewood, IllinoisGoogle Scholar
  44. Palant, C.E., Duffey, M.E., Mookerjee, B.K., Ho, S., Bentzel, C.J. 1983Ca2+ regulation of tight junction permeability and structure in Necturus gallbladderAm. J. Physiol.245C203C212PubMedGoogle Scholar
  45. Pitelka, D.R., Taggart, B.N., Hamamoto, S.T. 1983Effects of extracellular calcium depletion on membrane topography and occluding junctions of mammary epithelial cells in cultureJ. Cell Biol.96613624CrossRefPubMedGoogle Scholar
  46. Poncet, V., Merot, J., Poujeol, P. 1992A calcium-permeable channel in the apical membrane of primary cultures of the rabbit distal bright convoluted tubulePfluegers Archiv.422112119CrossRefPubMedGoogle Scholar
  47. Powis, D.A., Clark, C.L., O’Brien, K.J. 1994Lanthanum can be transported by the sodium-calcium exchange pathway and directly triggers catecholamine release from bovine chromaffin cellsCell Calcium16377390CrossRefPubMedGoogle Scholar
  48. Ramsey, C.R., Berndt, T.J., Knox, F.G. 1998Effect of volume expansion on the paracellular flux of lanthanum in the proximal tubuleJ. Am. Soc. Nephml.911471152Google Scholar
  49. Ramsey, C.R., Berndt, T.J., Knox, F.G. 2002Indomethacin blocks enhanced paracellular backflux in proximal tubulesJ. Am. Soc. Nephrol.1314491454CrossRefPubMedGoogle Scholar
  50. Ranscht, B. 1994Cadherins and catenins: interactions and functions in embryonic developmentCurr. Opin. Cell Biol.6740746CrossRefPubMedGoogle Scholar
  51. Ringwald, M., Schuh, R., Vestweber, D., Eistetter, H., Lottspeich, F., Engel, J., Dölz, R., Jähnig, F., Epplen, J., Mayer, S., Miiller, C., Kemler, R. 1987The structure of cell adhesion molecule uvomorulin. Insights into the molecular mechanism of Ca2+ dependent cell adhesionEMBO J.636473653PubMedGoogle Scholar
  52. Schneeberger, E., Lynch, R.D. 2004The tight junction: a multifunctional complexAm. J. Physiol.286C1213C1228CrossRefGoogle Scholar
  53. Sedar, A.W., Forte, J.G. 1964Effects of calcium depletion on the junctional complex between oxyntic cells of gastric glandsJ. Cell Biol.22173188CrossRefPubMedGoogle Scholar
  54. Shapiro, L., Fannon, A.M., Kwong, P.D., Thompson, A., Lehmann, M.S., Grubel, G., Legrand, J.F., Als-Nielsen, J., Colman, D.R., Hendrickson, W.A. 1995Structural basis of cell-cell adhesion by cadherinsNature374327337CrossRefPubMedGoogle Scholar
  55. Snyder, E., Buoscio, B.W., Falke, J.J. 1990Calcium(II) site specificity: Effect of size and charge on metal ion binding to an EF-hand-like siteBiochemistry2939373943CrossRefPubMedGoogle Scholar
  56. Suzuki, K., Kokai, Y., Sawada, N., Takakuwa, R., Kuwahara, K., Isogai, E., Isogai, H., Mori, M. 2002SS1 Helicobacter pylori disrupts the paracellular barrier of the gastric mucosa and leads to neutrophilic gastritis in miceVirchows Arch.440318324CrossRefPubMedGoogle Scholar
  57. Takata, M., Pickard, W.F., Lettvin, J.Y., Moore, J.W. 1966Ionic conductance changes in lobster axon membranes when lanthanum is substituted for calciumJ. Gen. Physiol.50461471CrossRefPubMedGoogle Scholar
  58. Takeichi, M. 1990Cadherins: A molecular family important in selective cell-cell adhesionAnnu. Rev. Biochem.59237252CrossRefPubMedGoogle Scholar
  59. Tomschy, A., Fauser, C., Landwehr, R., Engel, J. 1996Homophilic adhesion of E-cadherin occurs by a co-operative two- step interaction of N-terminal domainsEMBO J.1535073514PubMedGoogle Scholar
  60. Uehara, K., Miyoshi, M. 1999Tight junction of sinus endothelial cells of the rat spleenTissue Cell31555560CrossRefPubMedGoogle Scholar
  61. Vogel, W. 1974Calcium and lanthanum effects at the nodal membranePfluegers Archiv.3502539CrossRefPubMedGoogle Scholar
  62. Watkins, C.S., Mathie, A. 1994Modulation of the gating of the transient outward potassium current of rat isolated cerebellar granule neurons by lanthanumPfluegers Archiv.428209216CrossRefPubMedGoogle Scholar
  63. Weiss, D.J., Beckett, T., Bonneau, L., Young, J., Kolls, J.K., Wang, G. 2003Transient increase in lung epithelial tight junction permeability: an additional mechanism for enhancement of lung transgene expression by perfluorochemical liquidsMol. Ther.8927935CrossRefPubMedGoogle Scholar
  64. Weiss, G.B. 1974Cellular Pharmacology of LanthanumAnnu. Rev. Pharmacol.14343354CrossRefGoogle Scholar
  65. Yap, A.S., Brieher, W.M., Pruschy, M., Gumbiner, B.M. 1997Lateral clustering of the adhesive ectodomain: a fundamental determinant of cadherin functionCurr. Biol.7308315CrossRefPubMedGoogle Scholar
  66. Zheng, K., Makagiansar, I.T., Wang, M., Urbauer, J.L., Kuczera, K., Siahaan, T.J. 2004Expression, purification, and structural study of the EC4 domain of E-cadherinProtein Expr. Purif.337279CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  1. 1.Department of Physiology and Biophysics, Institute of Biomedical SciencesUniversity of São PauloSão PauloBrazil
  2. 2.Department of Operative Dentistry, School of DentistryUniversity of São PauloSão PauloBrazil

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