Experimental modulation of occluding junctions in a cultured transporting epithelium

doi:10.1083/jcb.87.3.736

This Article

	Full Text (PDF, 1161K)
	Alert me when this article is cited
	Citation Map

Services

	Email this article
	Similar articles in this journal
	Similar articles in PubMed
	Alert me to new content in the JCB
	Download to citation manager

Citing Articles

	Citing Articles via HighWire
	Citing Articles via CrossRef
	Citing Articles via Google Scholar

Google Scholar

	Articles by Martinez-Palomo, A.
	Articles by Cereijido, M.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Martinez-Palomo, A.
	Articles by Cereijido, M.


Social Bookmarking

	What's this?

ARTICLES

Experimental modulation of occluding junctions in a cultured transporting epithelium

A Martinez-Palomo, I Meza, G Beaty and M Cereijido

The experimental opening and resealing of occluding junctions in monolayers of cultured MDCK cells (epithelioid of renal origin) was explored by measuring changes in the electrical resistance across the monolayer and by freeze-fracture electron microscopy. As in natural epithelia, the function of occluding junctions as permeability barriers specifically depends on extracellular Ca++ concentration and fails if this ion is replaced by Mg++ or Ba++. The removal of Ca++ and the addition of EGTA to the bathing medium opened the junctions and reduced the transepithelial resistance. Resealing was achieved within 10-15 min by restoring Ca++. Quantitative freeze-fracture electron microscopy showed that junctional opening, caused by lack of Ca++, was accompanied by simplification of the pattern of the membrane strands of the occluding junction without disassembly or displacement of the junctional components. Resealing of the cellular contacts involved the gradual return to a normal junctional pattern estimated as the average number of strands constituting the junction. The occluding junctions were also opened by the addition of the ionophore A23187, suggesting that the sealing of the contacts requires high Ca++ on the extracellular side and low Ca++ concentration of the cytoplasmic compartment. The opening process could be blocked by low temperature (7.5 degrees C). Resealing did not depend on serum factors and did not require protein synthesis; therefore, it seems to be caused by reassembly of preexisting membrane junctional components. The restoration of the junctions occurred simultaneously with the establishment of ion-selective channels; the Na+/Cl- and the cation/cation selectivity were recovered with the same time-course as the electrical resistance. The role of the cytoskeleton in the process of junctional reassembly is reported in the companion article.
Add to CiteULike CiteULike Add to Complore Complore Add to Connotea Connotea Add to Del.icio.us Del.icio.us Add to Digg Digg Facebook Add to Reddit Reddit Add to Technorati Technorati Twitter What's this?

This article has been cited by other articles:

Wolburg, H., Noell, S., Wolburg-Buchholz, K., Mack, A., Fallier-Becker, P. (2009). Agrin, Aquaporin-4, and Astrocyte Polarity as an Important Feature of the Blood-Brain Barrier. Neuroscientist 15: 180-193 [Abstract]
Flynn, A. N., Itani, O. A., Moninger, T. O., Welsh, M. J. (2009). Acute regulation of tight junction ion selectivity in human airway epithelia. Proc. Natl. Acad. Sci. USA 106: 3591-3596 [Abstract] [Full Text]
Carrozzino, F., Pugnale, P., Feraille, E., Montesano, R. (2009). Inhibition of basal p38 or JNK activity enhances epithelial barrier function through differential modulation of claudin expression. Am. J. Physiol. Cell Physiol. 297: C775-C787 [Abstract] [Full Text]
Krautkramer, E., Zeier, M. (2008). Hantavirus Causing Hemorrhagic Fever with Renal Syndrome Enters from the Apical Surface and Requires Decay-Accelerating Factor (DAF/CD55). J. Virol. 82: 4257-4264 [Abstract] [Full Text]
Bolivar, J. J., Tapia, D., Arenas, G., Castanon-Arreola, M., Torres, H., Galarraga, E. (2008). A hyperpolarization-activated, cyclic nucleotide-gated, (Ih-like) cationic current and HCN gene expression in renal inner medullary collecting duct cells. Am. J. Physiol. Cell Physiol. 294: C893-C906 [Abstract] [Full Text]
Herman, R. E., Makienko, E. G., Prieve, M. G., Fuller, M., Houston, M. E. Jr, Johnson, P. H. (2007). Phage Display Screening of Epithelial Cell Monolayers Treated with EGTA: Identification of Peptide FDFWITP that Modulates Tight Junction Activity. J Biomol Screen 12: 1092-1101 [Abstract]
Carrozzino, F., Soulie, P., Huber, D., Mensi, N., Orci, L., Cano, A., Feraille, E., Montesano, R. (2005). Inducible expression of Snail selectively increases paracellular ion permeability and differentially modulates tight junction proteins. Am. J. Physiol. Cell Physiol. 289: C1002-C1014 [Abstract] [Full Text]
Nava, P., Lopez, S., Arias, C. F., Islas, S., Gonzalez-Mariscal, L. (2004). The rotavirus surface protein VP8 modulates the gate and fence function of tight junctions in epithelial cells. J. Cell Sci. 117: 5509-5519 [Abstract] [Full Text]
Cereijido, M., Contreras, R. G., Shoshani, L. (2004). Cell Adhesion, Polarity, and Epithelia in the Dawn of Metazoans. Physiol. Rev. 84: 1229-1262 [Abstract] [Full Text]
Lee, D. B. N., Jamgotchian, N., Allen, S. G., Kan, F. W. K., Hale, I. L. (2004). Annexin A2 heterotetramer: role in tight junction assembly. Am. J. Physiol. Renal Physiol. 287: F481-F491 [Abstract] [Full Text]
Bazzoni, G., Dejana, E. (2004). Endothelial Cell-to-Cell Junctions: Molecular Organization and Role in Vascular Homeostasis. Physiol. Rev. 84: 869-901 [Abstract] [Full Text]
Tobey, N. A., Argote, C. M., Hosseini, S. S., Orlando, R. C. (2004). Calcium-switch technique and junctional permeability in native rabbit esophageal epithelium. Am. J. Physiol. Gastrointest. Liver Physiol. 286: G1042-G1049 [Abstract] [Full Text]
Muza-Moons, M. M., Koutsouris, A., Hecht, G. (2003). Disruption of Cell Polarity by Enteropathogenic Escherichia coli Enables Basolateral Membrane Proteins To Migrate Apically and To Potentiate Physiological Consequences. Infect. Immun. 71: 7069-7078 [Abstract] [Full Text]
Watson, C. J., Rowland, M., Warhurst, G. (2001). Functional modeling of tight junctions in intestinal cell monolayers using polyethylene glycol oligomers. Am. J. Physiol. Cell Physiol. 281: C388-C397 [Abstract] [Full Text]
Hamm-Alvarez, S. F., Chang, A., Wang, Y., Jerdeva, G., Lin, H. H., Kim, K.-J., Ann, D. K. (2001). Etk/Bmx activation modulates barrier function in epithelial cells. Am. J. Physiol. Cell Physiol. 280: C1657-C1668 [Abstract] [Full Text]
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. JCB 153: 263-272 [Abstract] [Full Text]
Yang, X.-Y., Zhao, H., Zhang, Z., Rodland, K. D., Roullet, J.-B., Cohen, D. M. (2001). Urea signaling to ERK phosphorylation in renal medullary cells requires extracellular calcium but not calcium entry. Am. J. Physiol. Renal Physiol. 280: F162-F171 [Abstract] [Full Text]
Goto, Y., Uchida, Y., Nomura, A., Sakamoto, T., Ishii, Y., Morishima, Y., Masuyama, K., Sekizawa, K. (2000). Dislocation of E-Cadherin in the Airway Epithelium during an Antigen-Induced Asthmatic Response. Am. J. Respir. Cell Mol. Bio. 23: 712-718 [Abstract] [Full Text]
Cereijido, M., Shoshani, L., Contreras, R. G. (2000). Molecular Physiology and Pathophysiology of Tight Junctions I. Biogenesis of tight junctions and epithelial polarity. Am. J. Physiol. Gastrointest. Liver Physiol. 279: G477-G482 [Abstract] [Full Text]
Hileman, S. M., Tornoe, J., Flier, J. S., Bjorbak, C. (2000). Transcellular Transport of Leptin by the Short Leptin Receptor Isoform ObRa in Madin-Darby Canine Kidney Cells. Endocrinology 141: 1955-1961 [Abstract] [Full Text]
Renold, A., Cescato, R., Beuret, N., Vogel, L. K., Wahlberg, J. M., Brown, J. L., Fiedler, K., Spiess, M. (2000). Basolateral Sorting Signals Differ in Their Ability to Redirect Apical Proteins to the Basolateral Cell Surface. J. Biol. Chem. 275: 9290-9295 [Abstract] [Full Text]
Clarke, H, Soler, A., Mullin, J. (2000). Protein kinase C activation leads to dephosphorylation of occludin and tight junction permeability increase in LLC-PK1 epithelial cell sheets. J. Cell Sci. 113: 3187-3196 [Abstract]
Liu, Y, Nusrat, A, Schnell, F., Reaves, T., Walsh, S, Pochet, M, Parkos, C. (2000). Human junction adhesion molecule regulates tight junction resealing in epithelia. J. Cell Sci. 113: 2363-2374 [Abstract]
Walters, R. W., Grunst, T., Bergelson, J. M., Finberg, R. W., Welsh, M. J., Zabner, J. (1999). Basolateral Localization of Fiber Receptors Limits Adenovirus Infection from the Apical Surface of Airway Epithelia. J. Biol. Chem. 274: 10219-10226 [Abstract] [Full Text]
Grima, J., Wong, C. C.�S., Zhu, L.-j., Zong, S.-d., Cheng, C. Y. (1998). Testin Secreted by Sertoli Cells Is Associated with the Cell Surface, and Its Expression Correlates with the Disruption of Sertoli-Germ Cell Junctions but Not the Inter-Sertoli Tight Junction. J. Biol. Chem. 273: 21040-21053 [Abstract] [Full Text]
Kloub, M. A., Heck, G. L., Desimone, J. A. (1998). Self-Inhibition in Ca2+-Evoked Taste Responses: A Novel Tool for Functional Dissection of Salt Taste Transduction Mechanisms. J. Neurophysiol. 79: 911-921 [Abstract] [Full Text]
Lacaz-Vieira, F. (1997). Calcium Site Specificity: Early Ca2+-related Tight Junction Events. JGP 110: 727-740 [Abstract] [Full Text]
Marquis, R. E., Hudspeth, A. J. (1997). Effects of extracellular Ca2+ concentration on hair-bundle stiffness and gating-spring integrity in hair�cells. Proc. Natl. Acad. Sci. USA 94: 11923-11928 [Abstract] [Full Text]
Lumpkin, E. A., Marquis, R. E., Hudspeth, A. J. (1997). The selectivity of the hair cell's mechanoelectrical-transduction channel promotes Ca2+ flux at low Ca2+ concentrations. Proc. Natl. Acad. Sci. USA 94: 10997-11002 [Abstract] [Full Text]
Ingemansson, R., Bolys, R., Budrikis, A., Lindgren, A., Sjoberg, T., Steen, S. (1997). Addition of Calcium to Euro-Collins Solution Is Essential for 24-Hour Preservation of the Vasculature. Ann. Thorac. Surg. 63: 408-413 [Abstract] [Full Text]
Staddon, J., Herrenknecht, K, Smales, C, Rubin, L. (1995). Evidence that tyrosine phosphorylation may increase tight junction permeability. J. Cell Sci. 108: 609-619 [Abstract]
Stevenson, B., Begg, D. (1994). Concentration-dependent effects of cytochalasin D on tight junctions and actin filaments in MDCK epithelial cells. J. Cell Sci. 107: 367-375 [Abstract]
Mazariegos, M.R., Hand, A.R. (1984). Basic Biological Sciences Regulation of Tight Junctional Permeability in the Rat Parotid Gland by Autonomic Agonists. JDR 63: 1102-1107 [Abstract]