Key points are not available for this paper at this time.
A key feature of the ischemic epithelial cell phenotype is the disruption of tight junctions (TJ). In a Manin-Darby canine kidney cell model for ischemia-reperfusion/hypoxia-reoxygenation injury which employs inhibitors of glycolysis (2-deoxy-d-glucose) and oxidative phosphorylation (antimycin A), transepithelial electrical resistance, a measure of TJ integrity, dropped rapidly, correlating well with declining ATP levels. Although immunocytochemical studies revealed only subtle changes in the distribution of the TJ proteins, zonula occludens (ZO)-1, ZO-2, and cingulin, examination of the Triton X-100 solubilities of these proteins, an indicator of cytoskeletal association, revealed a striking shift of all three TJ proteins into the insoluble pool, consistent with increased cytoskeletal interaction during ATP depletion. In addition, rate-zonal centrifugation analysis of a detergent-soluble fraction showed an increase in the amount of ZO-1 and ZO-2 in high density fractions following ATP depletion, providing further evidence for association of TJ proteins into a large complex possibly involving the cytoskeleton. Analysis of immunoprecipitation data from 35Smethionine-labeled cells revealed that ATP depletion led to the association of a 240-kDa protein with the ZO-1-containing complex. Western blots of this protein immunoprecipitated with anti-ZO-1 antibodies confirmed its identity as fodrin, a protein believed to link membrane and other proteins to the actin-based cytoskeleton. Together, our data suggest that in the absence of major immunocytochemical changes, ATP depletion leads TJ proteins to form large insoluble complexes and associate with the cytoskeleton. We propose a model in which a key, potentially regulated, step in the generation of the ischemic epithelial cell phenotype is the interaction between TJ proteins and fodrin and/or other cytoskeletal proteins. A key feature of the ischemic epithelial cell phenotype is the disruption of tight junctions (TJ). In a Manin-Darby canine kidney cell model for ischemia-reperfusion/hypoxia-reoxygenation injury which employs inhibitors of glycolysis (2-deoxy-d-glucose) and oxidative phosphorylation (antimycin A), transepithelial electrical resistance, a measure of TJ integrity, dropped rapidly, correlating well with declining ATP levels. Although immunocytochemical studies revealed only subtle changes in the distribution of the TJ proteins, zonula occludens (ZO)-1, ZO-2, and cingulin, examination of the Triton X-100 solubilities of these proteins, an indicator of cytoskeletal association, revealed a striking shift of all three TJ proteins into the insoluble pool, consistent with increased cytoskeletal interaction during ATP depletion. In addition, rate-zonal centrifugation analysis of a detergent-soluble fraction showed an increase in the amount of ZO-1 and ZO-2 in high density fractions following ATP depletion, providing further evidence for association of TJ proteins into a large complex possibly involving the cytoskeleton. Analysis of immunoprecipitation data from 35Smethionine-labeled cells revealed that ATP depletion led to the association of a 240-kDa protein with the ZO-1-containing complex. Western blots of this protein immunoprecipitated with anti-ZO-1 antibodies confirmed its identity as fodrin, a protein believed to link membrane and other proteins to the actin-based cytoskeleton. Together, our data suggest that in the absence of major immunocytochemical changes, ATP depletion leads TJ proteins to form large insoluble complexes and associate with the cytoskeleton. We propose a model in which a key, potentially regulated, step in the generation of the ischemic epithelial cell phenotype is the interaction between TJ proteins and fodrin and/or other cytoskeletal proteins. The epithelial intercellular permeability barrier is maintained largely by the tight junction (TJ) 1The abbreviations used are: TJ, tight junction; ZO-1, zonula occludens 1; ZO-2, zonula occludens 2; MDCK, Madin-Darby canine kidney; TER, transepithelial electrical resistance; PBS, phosphate-buffered saline; TRITC, tetramethylrhodamine isothiocyanate; PAGE, polyacrylamide gel electrophoresis. 1The abbreviations used are: TJ, tight junction; ZO-1, zonula occludens 1; ZO-2, zonula occludens 2; MDCK, Madin-Darby canine kidney; TER, transepithelial electrical resistance; PBS, phosphate-buffered saline; TRITC, tetramethylrhodamine isothiocyanate; PAGE, polyacrylamide gel electrophoresis. (1Anderson J.M. Balda M.S. Fanning A.S. Curr. Opin. Cell Biol. 1993; 5: 772-778Crossref PubMed Scopus (181) Google Scholar). The TJ, the most apical of intercellular junctions, consists of a number of proteins, including ZO-1, ZO-2, occludin, cingulin, 7H6, p130, and potentially other proteins (2Citi S. J. Cell Biol. 1993; 121: 485-489Crossref PubMed Scopus (169) Google Scholar, 3Itoh M. Nagafuchi A. Yonemura S. Kitani-Yasuda T. Tsukita S. Tsukita S. J. Cell Biol. 1993; 121: 491-502Crossref PubMed Scopus (496) Google Scholar, 4Willott E. Balda M.S. Fanning A.S. Jameson B. Itallie C.V. Anderson J.M. Proc. Natl. Acad. Sci. U. S. A. 1993; 90: 7834-7838Crossref PubMed Scopus (422) Google Scholar, 5Jesaitis L.A. Goodenough D.A. J. Cell Biol. 1994; 124: 949-961Crossref PubMed Scopus (386) Google Scholar, 6Zhong Y. Saitoh T. Minase T. Sawada N. Enomoto K. Mori M. J. Cell Biol. 1993; 120: 477-483Crossref PubMed Scopus (244) Google Scholar, 7Citi S. Sabanay H. Jakes R. Geiger B. Kendrick-Jones J. Nature. 1988; 333: 272-276Crossref PubMed Scopus (401) Google Scholar, 8Stuart R.O. Nigam S.K. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 6072-6076Crossref PubMed Scopus (242) Google Scholar, 9Keon B.H. Schäfer S. Kuhn C. Grund C. Franke W.W. J. Cell Biol. 1996; 134: 1003-1018Crossref PubMed Scopus (267) Google Scholar). Considerable indirect evidence suggests that proteins of the TJ are intimately associated with the actin-based cytoskeleton (10Madara J.L. Am. J. Physiol. 1987; 253: C171-C175Crossref PubMed Google Scholar, 11Itoh M. Yonemura S. Nagafuchi A. Tsukita S. Tsukita S. J. Cell Biol. 1991; 115: 1449-1462Crossref PubMed Scopus (204) Google Scholar, 12Furuse M. Itoh M. Hirase T. Nagafuchi A. Yonemura S. Tsukita S. Tsukita S. J. Cell Biol. 1994; 127: 1617-1626Crossref PubMed Scopus (798) Google Scholar).Ischemia and subsequent reperfusion/reoxygenation causes a number of lesions in epithelial cells including mispolarization of at least some membrane proteins, perturbation of the actin cytoskeleton, and disruption of the permeability barrier (13Molitoris B.A. Nelson W.J. J. Clin. Invest. 1990; 85: 3-9Crossref PubMed Scopus (95) Google Scholar, 14Fish E.M. Molitoris B.A. N. Engl. J. Med. 1994; 330: 1580-1587Crossref PubMed Scopus (193) Google Scholar). These lesions have been reproduced in cell culture models for hypoxia-reoxygenation injury using agents that deplete cellular ATP, which has allowed for the analysis of molecular mechanisms underlying ischemic injury (15Canfield P.E. Geerdes A.M. Molitoris B.A. Am. J. Physiol. 1991; 261: F1038-F1045PubMed Google Scholar, 16Mandel L.J. Bacallao R. Zampighi G. Nature. 1993; 361: 552-555Crossref PubMed Scopus (233) Google Scholar). Although mechanistic insights into the disruption of the actin-based cytoskeleton are beginning to emerge, little is known about the biochemical basis of the disruption of the TJ after ischemic insult or how the TJ reassembles during recovery of epithelial cells from ischemic injury.The biochemical basis of the disassembly and reassembly of the TJ has, however, been studied in MDCK cells in a model in which extracellular calcium is manipulated: the “calcium switch” (8Stuart R.O. Nigam S.K. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 6072-6076Crossref PubMed Scopus (242) Google Scholar, 17Nigam S.K. Brenner B.M. Curr. Opin. Nephrol. Hypertension. 1992; 1: 187-191Crossref PubMed Scopus (5) Google Scholar, 18Stuart R.O. Sun A. Panichas M. Hebert S.C. Brenner B.M. Nigam S.K. J. Cell. Physiol. 1994; 159: 423-433Crossref PubMed Scopus (105) Google Scholar, 19Stuart R.O. Sun A. Bush K.T. Nigam S.K. J. Biol. Chem. 1996; 271: 13636-13641Abstract PubMed Scopus Google Scholar, B.M. C. S. Nigam S.K. J. Biol. Chem. 1996; 271: PubMed Scopus Google Scholar). the TJ in this model calcium transepithelial electrical is and TJ proteins or are the cell In this TJ proteins with a of with the cytoskeleton, and ZO-1, ZO-2, and are in a complex which other (8Stuart R.O. Nigam S.K. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 6072-6076Crossref PubMed Scopus (242) Google Scholar). extracellular calcium is reassembly of the TJ to by involving a calcium R.O. Sun A. Panichas M. Hebert S.C. Brenner B.M. Nigam S.K. J. Cell. Physiol. 1994; 159: 423-433Crossref PubMed Scopus (105) Google Scholar, 19Stuart R.O. Sun A. Bush K.T. Nigam S.K. J. Biol. Chem. 1996; 271: 13636-13641Abstract PubMed Scopus Google Scholar, B.M. C. S. Nigam S.K. J. Biol. Chem. 1996; 271: PubMed Scopus Google Scholar, S.K. E. Proc. Natl. Acad. Sci. U. S. A. 1992; PubMed Scopus Google and protein (8Stuart R.O. Nigam S.K. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 6072-6076Crossref PubMed Scopus (242) Google Scholar). TJ proteins to the of the and TJ proteins to (8Stuart R.O. Nigam S.K. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 6072-6076Crossref PubMed Scopus (242) Google Scholar, 18Stuart R.O. Sun A. Panichas M. Hebert S.C. Brenner B.M. Nigam S.K. J. Cell. Physiol. 1994; 159: 423-433Crossref PubMed Scopus (105) Google Scholar, 19Stuart R.O. Sun A. Bush K.T. Nigam S.K. J. Biol. Chem. 1996; 271: 13636-13641Abstract PubMed Scopus Google is known about the of the TJ in MDCK after ATP depletion and the disassembly of the TJ in calcium and its reassembly calcium is in cell culture models for these have been in the biochemical as with extracellular calcium and the calcium and biochemical have the of TJ proteins in a MDCK cell model for hypoxia-reoxygenation which employs A and (15Canfield P.E. Geerdes A.M. Molitoris B.A. Am. J. Physiol. 1991; 261: F1038-F1045PubMed Google Scholar, Bacallao R. L.J. Am. J. Physiol. 1994; PubMed Google and this model with the calcium We that the biochemical changes during TJ disassembly after ATP depletion are from cells are to calcium In this TJ disassembly is by the association of TJ proteins into large of at least three TJ proteins into an insoluble pool, and an increased association of the ZO-1-containing complex with the membrane cytoskeletal insights into the molecular of epithelial from ischemic studies of in and ATP have a disruption of the actin-based cytoskeleton E.M. Molitoris B.A. N. Engl. J. Med. 1994; 330: 1580-1587Crossref PubMed Scopus (193) Google Scholar, R. A. S. Zampighi G. L.J. J. Cell Sci. 1994; PubMed Google Scholar, A.M. Am. J. Physiol. 1994; Google Scholar, Nelson W.J. 1995; PubMed Scopus Google Scholar, S. K. M. A. B. J.M. Am. J. Physiol. 1996; Scholar). to of membrane proteins and and of the permeability barrier in epithelial cells (13Molitoris B.A. Nelson W.J. J. Clin. Invest. 1990; 85: 3-9Crossref PubMed Scopus (95) Google Scholar, 14Fish E.M. Molitoris B.A. N. Engl. J. Med. 1994; 330: 1580-1587Crossref PubMed Scopus (193) Google Scholar, 16Mandel L.J. Bacallao R. Zampighi G. Nature. 1993; 361: 552-555Crossref PubMed Scopus (233) Google Scholar). the TJ with the cytoskeleton, the of after ATP depletion by and for by cytoskeletal (1Anderson J.M. Balda M.S. Fanning A.S. Curr. Opin. Cell Biol. 1993; 5: 772-778Crossref PubMed Scopus (181) Google Scholar, 11Itoh M. Yonemura S. Nagafuchi A. Tsukita S. Tsukita S. J. Cell Biol. 1991; 115: 1449-1462Crossref PubMed Scopus (204) Google Scholar, 12Furuse M. Itoh M. Hirase T. Nagafuchi A. Yonemura S. Tsukita S. Tsukita S. J. Cell Biol. 1994; 127: 1617-1626Crossref PubMed Scopus (798) Google Scholar). the mechanisms of TJ disassembly underlying ATP depletion in epithelial an interaction between TJ proteins and cytoskeleton and In these have that after the in which with a in cellular ATP three TJ proteins ZO-2, and insoluble and and that are in large molecular complexes all three TJ proteins largely to the of the membrane ATP depletion leads the ZO-1-containing complex of TJ proteins to or with the cytoskeletal fodrin and of these changes to largely after of ATP depletion and suggest that of the actin-based cytoskeleton after ischemic insult and ATP depletion calcium changes S. K. M. A. B. J.M. Am. J. Physiol. 1996; Scholar, J.M. J. Clin. Invest. 1991; PubMed Scopus Google Scholar, R. J.M. J. Am. J. Physiol. 1993; Google Scholar, A. P.E. J. Clin. Invest. 1994; PubMed Scopus Google Scholar). is that these changes in calcium to the changes in biochemical of TJ proteins and cytoskeletal association which have after ATP depletion. a high calcium is to an association, at least in which the TJ is calcium is with or calcium in the absence of ATP depletion, is increase in the of TJ proteins changes in calcium an in TJ after TJ disassembly in the of ATP the a in the of these during reassembly after ATP is for a in the calcium changes in calcium have been at least to the of TJ proteins R.O. Sun A. Bush K.T. Nigam S.K. J. Biol. Chem. 1996; 271: 13636-13641Abstract PubMed Scopus Google Scholar). In this model for TJ disassembly and the reassembly of the TJ a protein B.M. C. S. Nigam S.K. J. Biol. Chem. 1996; 271: PubMed Scopus Google calcium R.O. Sun A. Panichas M. Hebert S.C. Brenner B.M. Nigam S.K. J. Cell. Physiol. 1994; 159: 423-433Crossref PubMed Scopus (105) Google Scholar, 19Stuart R.O. Sun A. Bush K.T. Nigam S.K. J. Biol. Chem. 1996; 271: 13636-13641Abstract PubMed Scopus Google and protein (8Stuart R.O. Nigam S.K. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 6072-6076Crossref PubMed Scopus (242) Google Scholar). to in studies the reassembly of the TJ after ATP is a to link a number of membrane proteins to the cytoskeleton W.J. J. Cell Biol. PubMed Scopus Google Scholar, W.J. E.M. J. Cell Biol. 1990; PubMed Scopus (244) Google Scholar, S. Tsukita S. Nagafuchi A. Yonemura S. Curr. Opin. Cell Biol. 1992; PubMed Scopus Google Scholar). as well as has been to with proteins in epithelial cells Physiol. 1990; Scopus Google Scholar, Nelson W.J. Am. J. Physiol. 1996; PubMed Google Scholar). data that after ATP depletion, the fraction of fodrin the TJ Although to fodrin with the ZO-1-containing this association is or our that the of between the TJ and cytoskeleton is in the of epithelial cell ischemic and propose that a key step in this is the association of TJ proteins with fodrin and/or other cytoskeletal proteins, an association that by mechanisms involving calcium and/or protein that with this association of in the of ischemic The epithelial intercellular permeability barrier is maintained largely by the tight junction (TJ) 1The abbreviations used are: TJ, tight junction; ZO-1, zonula occludens 1; ZO-2, zonula occludens 2; MDCK, Madin-Darby canine kidney; TER, transepithelial electrical resistance; PBS, phosphate-buffered saline; TRITC, tetramethylrhodamine isothiocyanate; PAGE, polyacrylamide gel electrophoresis. 1The abbreviations used are: TJ, tight junction; ZO-1, zonula occludens 1; ZO-2, zonula occludens 2; MDCK, Madin-Darby canine kidney; TER, transepithelial electrical resistance; PBS, phosphate-buffered saline; TRITC, tetramethylrhodamine isothiocyanate; PAGE, polyacrylamide gel electrophoresis. (1Anderson J.M. Balda M.S. Fanning A.S. Curr. Opin. Cell Biol. 1993; 5: 772-778Crossref PubMed Scopus (181) Google Scholar). The TJ, the most apical of intercellular junctions, consists of a number of proteins, including ZO-1, ZO-2, occludin, cingulin, 7H6, p130, and potentially other proteins (2Citi S. J. Cell Biol. 1993; 121: 485-489Crossref PubMed Scopus (169) Google Scholar, 3Itoh M. Nagafuchi A. Yonemura S. Kitani-Yasuda T. Tsukita S. Tsukita S. J. Cell Biol. 1993; 121: 491-502Crossref PubMed Scopus (496) Google Scholar, 4Willott E. Balda M.S. Fanning A.S. Jameson B. Itallie C.V. Anderson J.M. Proc. Natl. Acad. Sci. U. S. A. 1993; 90: 7834-7838Crossref PubMed Scopus (422) Google Scholar, 5Jesaitis L.A. Goodenough D.A. J. Cell Biol. 1994; 124: 949-961Crossref PubMed Scopus (386) Google Scholar, 6Zhong Y. Saitoh T. Minase T. Sawada N. Enomoto K. Mori M. J. Cell Biol. 1993; 120: 477-483Crossref PubMed Scopus (244) Google Scholar, 7Citi S. Sabanay H. Jakes R. Geiger B. Kendrick-Jones J. Nature. 1988; 333: 272-276Crossref PubMed Scopus (401) Google Scholar, 8Stuart R.O. Nigam S.K. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 6072-6076Crossref PubMed Scopus (242) Google Scholar, 9Keon B.H. Schäfer S. Kuhn C. Grund C. Franke W.W. J. Cell Biol. 1996; 134: 1003-1018Crossref PubMed Scopus (267) Google Scholar). Considerable indirect evidence suggests that proteins of the TJ are intimately associated with the actin-based cytoskeleton (10Madara J.L. Am. J. Physiol. 1987; 253: C171-C175Crossref PubMed Google Scholar, 11Itoh M. Yonemura S. Nagafuchi A. Tsukita S. Tsukita S. J. Cell Biol. 1991; 115: 1449-1462Crossref PubMed Scopus (204) Google Scholar, 12Furuse M. Itoh M. Hirase T. Nagafuchi A. Yonemura S. Tsukita S. Tsukita S. J. Cell Biol. 1994; 127: 1617-1626Crossref PubMed Scopus (798) Google Scholar). and subsequent reperfusion/reoxygenation causes a number of lesions in epithelial cells including mispolarization of at least some membrane proteins, perturbation of the actin cytoskeleton, and disruption of the permeability barrier (13Molitoris B.A. Nelson W.J. J. Clin. Invest. 1990; 85: 3-9Crossref PubMed Scopus (95) Google Scholar, 14Fish E.M. Molitoris B.A. N. Engl. J. Med. 1994; 330: 1580-1587Crossref PubMed Scopus (193) Google Scholar). These lesions have been reproduced in cell culture models for hypoxia-reoxygenation injury using agents that deplete cellular ATP, which has allowed for the analysis of molecular mechanisms underlying ischemic injury (15Canfield P.E. Geerdes A.M. Molitoris B.A. Am. J. Physiol. 1991; 261: F1038-F1045PubMed Google Scholar, 16Mandel L.J. Bacallao R. Zampighi G. Nature. 1993; 361: 552-555Crossref PubMed Scopus (233) Google Scholar). Although mechanistic insights into the disruption of the actin-based cytoskeleton are beginning to emerge, little is known about the biochemical basis of the disruption of the TJ after ischemic insult or how the TJ reassembles during recovery of epithelial cells from ischemic The biochemical basis of the disassembly and reassembly of the TJ has, however, been studied in MDCK cells in a model in which extracellular calcium is manipulated: the “calcium switch” (8Stuart R.O. Nigam S.K. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 6072-6076Crossref PubMed Scopus (242) Google Scholar, 17Nigam S.K. Brenner B.M. Curr. Opin. Nephrol. Hypertension. 1992; 1: 187-191Crossref PubMed Scopus (5) Google Scholar, 18Stuart R.O. Sun A. Panichas M. Hebert S.C. Brenner B.M. Nigam S.K. J. Cell. Physiol. 1994; 159: 423-433Crossref PubMed Scopus (105) Google Scholar, 19Stuart R.O. Sun A. Bush K.T. Nigam S.K. J. Biol. Chem. 1996; 271: 13636-13641Abstract PubMed Scopus Google Scholar, B.M. C. S. Nigam S.K. J. Biol. Chem. 1996; 271: PubMed Scopus Google Scholar). the TJ in this model calcium transepithelial electrical is and TJ proteins or are the cell In this TJ proteins with a of with the cytoskeleton, and ZO-1, ZO-2, and are in a complex which other (8Stuart R.O. Nigam S.K. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 6072-6076Crossref PubMed Scopus (242) Google Scholar). extracellular calcium is reassembly of the TJ to by involving a calcium R.O. Sun A. Panichas M. Hebert S.C. Brenner B.M. Nigam S.K. J. Cell. Physiol. 1994; 159: 423-433Crossref PubMed Scopus (105) Google Scholar, 19Stuart R.O. Sun A. Bush K.T. Nigam S.K. J. Biol. Chem. 1996; 271: 13636-13641Abstract PubMed Scopus Google Scholar, B.M. C. S. Nigam S.K. J. Biol. Chem. 1996; 271: PubMed Scopus Google Scholar, S.K. E. Proc. Natl. Acad. Sci. U. S. A. 1992; PubMed Scopus Google and protein (8Stuart R.O. Nigam S.K. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 6072-6076Crossref PubMed Scopus (242) Google Scholar). TJ proteins to the of the and TJ proteins to (8Stuart R.O. Nigam S.K. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 6072-6076Crossref PubMed Scopus (242) Google Scholar, 18Stuart R.O. Sun A. Panichas M. Hebert S.C. Brenner B.M. Nigam S.K. J. Cell. Physiol. 1994; 159: 423-433Crossref PubMed Scopus (105) Google Scholar, 19Stuart R.O. Sun A. Bush K.T. Nigam S.K. J. Biol. Chem. 1996; 271: 13636-13641Abstract PubMed Scopus Google Scholar). is known about the of the TJ in MDCK after ATP depletion and the disassembly of the TJ in calcium and its reassembly calcium is in cell culture models for these have been in the biochemical as with extracellular calcium and the calcium and biochemical have the of TJ proteins in a MDCK cell model for hypoxia-reoxygenation which employs A and (15Canfield P.E. Geerdes A.M. Molitoris B.A. Am. J. Physiol. 1991; 261: F1038-F1045PubMed Google Scholar, Bacallao R. L.J. Am. J. Physiol. 1994; PubMed Google and this model with the calcium We that the biochemical changes during TJ disassembly after ATP depletion are from cells are to calcium In this TJ disassembly is by the association of TJ proteins into large of at least three TJ proteins into an insoluble pool, and an increased association of the ZO-1-containing complex with the membrane cytoskeletal insights into the molecular of epithelial from ischemic studies of in and ATP have a disruption of the actin-based cytoskeleton E.M. Molitoris B.A. N. Engl. J. Med. 1994; 330: 1580-1587Crossref PubMed Scopus (193) Google Scholar, R. A. S. Zampighi G. L.J. J. Cell Sci. 1994; PubMed Google Scholar, A.M. Am. J. Physiol. 1994; Google Scholar, Nelson W.J. 1995; PubMed Scopus Google Scholar, S. K. M. A. B. J.M. Am. J. Physiol. 1996; Scholar). to of membrane proteins and and of the permeability barrier in epithelial cells (13Molitoris B.A. Nelson W.J. J. Clin. Invest. 1990; 85: 3-9Crossref PubMed Scopus (95) Google Scholar, 14Fish E.M. Molitoris B.A. N. Engl. J. Med. 1994; 330: 1580-1587Crossref PubMed Scopus (193) Google Scholar, 16Mandel L.J. Bacallao R. Zampighi G. Nature. 1993; 361: 552-555Crossref PubMed Scopus (233) Google Scholar). the TJ with the cytoskeleton, the of after ATP depletion by and for by cytoskeletal (1Anderson J.M. Balda M.S. Fanning A.S. Curr. Opin. Cell Biol. 1993; 5: 772-778Crossref PubMed Scopus (181) Google Scholar, 11Itoh M. Yonemura S. Nagafuchi A. Tsukita S. Tsukita S. J. Cell Biol. 1991; 115: 1449-1462Crossref PubMed Scopus (204) Google Scholar, 12Furuse M. Itoh M. Hirase T. Nagafuchi A. Yonemura S. Tsukita S. Tsukita S. J. Cell Biol. 1994; 127: 1617-1626Crossref PubMed Scopus (798) Google Scholar). the mechanisms of TJ disassembly underlying ATP depletion in epithelial an interaction between TJ proteins and cytoskeleton and In these have that after the in which with a in cellular ATP three TJ proteins ZO-2, and insoluble and and that are in large molecular complexes all three TJ proteins largely to the of the membrane ATP depletion leads the ZO-1-containing complex of TJ proteins to or with the cytoskeletal fodrin and of these changes to largely after of ATP depletion and suggest that of the actin-based cytoskeleton after ischemic insult and ATP depletion calcium changes S. K. M. A. B. J.M. Am. J. Physiol. 1996; Scholar, J.M. J. Clin. Invest. 1991; PubMed Scopus Google Scholar, R. J.M. J. Am. J. Physiol. 1993; Google Scholar, A. P.E. J. Clin. Invest. 1994; PubMed Scopus Google Scholar). is that these changes in calcium to the changes in biochemical of TJ proteins and cytoskeletal association which have after ATP depletion. a high calcium is to an association, at least in which the TJ is calcium is with or calcium in the absence of ATP depletion, is increase in the of TJ proteins changes in calcium an in TJ after TJ disassembly in the of ATP the a in the of these during reassembly after ATP is for a in the calcium changes in calcium have been at least to the of TJ proteins R.O. Sun A. Bush K.T. Nigam S.K. J. Biol. Chem. 1996; 271: 13636-13641Abstract PubMed Scopus Google Scholar). In this model for TJ disassembly and the reassembly of the TJ a protein B.M. C. S. Nigam S.K. J. Biol. Chem. 1996; 271: PubMed Scopus Google calcium R.O. Sun A. Panichas M. Hebert S.C. Brenner B.M. Nigam S.K. J. Cell. Physiol. 1994; 159: 423-433Crossref PubMed Scopus (105) Google Scholar, 19Stuart R.O. Sun A. Bush K.T. Nigam S.K. J. Biol. Chem. 1996; 271: 13636-13641Abstract PubMed Scopus Google and protein (8Stuart R.O. Nigam S.K. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 6072-6076Crossref PubMed Scopus (242) Google Scholar). to in studies the reassembly of the TJ after ATP is a to link a number of membrane proteins to the cytoskeleton W.J. J. Cell Biol. PubMed Scopus Google Scholar, W.J. E.M. J. Cell Biol. 1990; PubMed Scopus (244) Google Scholar, S. Tsukita S. Nagafuchi A. Yonemura S. Curr. Opin. Cell Biol. 1992; PubMed Scopus Google Scholar). as well as has been to with proteins in epithelial cells Physiol. 1990; Scopus Google Scholar, Nelson W.J. Am. J. Physiol. 1996; PubMed Google Scholar). data that after ATP depletion, the fraction of fodrin the TJ Although to fodrin with the ZO-1-containing this association is or our that the of between the TJ and cytoskeleton is in the of epithelial cell ischemic and propose that a key step in this is the association of TJ proteins with fodrin and/or other cytoskeletal proteins, an association that by mechanisms involving calcium and/or protein that with this association of in the of ischemic studies of in and ATP have a disruption of the actin-based cytoskeleton E.M. Molitoris B.A. N. Engl. J. Med. 1994; 330: 1580-1587Crossref PubMed Scopus (193) Google Scholar, R. A. S. Zampighi G. L.J. J. Cell Sci. 1994; PubMed Google Scholar, A.M. Am. J. Physiol. 1994; Google Scholar, Nelson W.J. 1995; PubMed Scopus Google Scholar, S. K. M. A. B. J.M. Am. J. Physiol. 1996; Scholar). to of membrane proteins and and of the permeability barrier in epithelial cells (13Molitoris B.A. Nelson W.J. J. Clin. Invest. 1990; 85: 3-9Crossref PubMed Scopus (95) Google Scholar, 14Fish E.M. Molitoris B.A. N. Engl. J. Med. 1994; 330: 1580-1587Crossref PubMed Scopus (193) Google Scholar, 16Mandel L.J. Bacallao R. Zampighi G. Nature. 1993; 361: 552-555Crossref PubMed Scopus (233) Google Scholar). the TJ with the cytoskeleton, the of after ATP depletion by and for by cytoskeletal (1Anderson J.M. Balda M.S. Fanning A.S. Curr. Opin. Cell Biol. 1993; 5: 772-778Crossref PubMed Scopus (181) Google Scholar, 11Itoh M. Yonemura S. Nagafuchi A. Tsukita S. Tsukita S. J. Cell Biol. 1991; 115: 1449-1462Crossref PubMed Scopus (204) Google Scholar, 12Furuse M. Itoh M. Hirase T. Nagafuchi A. Yonemura S. Tsukita S. Tsukita S. J. Cell Biol. 1994; 127: 1617-1626Crossref PubMed Scopus (798) Google Scholar). the mechanisms of TJ disassembly underlying ATP depletion in epithelial an interaction between TJ proteins and cytoskeleton and In these have that after the in which with a in cellular ATP three TJ proteins ZO-2, and insoluble and and that are in large molecular complexes all three TJ proteins largely to the of the membrane ATP depletion leads the ZO-1-containing complex of TJ proteins to or with the cytoskeletal fodrin and of these changes to largely after of ATP depletion and suggest that of the actin-based cytoskeleton after ischemic insult and ATP depletion calcium changes S. K. M. A. B. J.M. Am. J. Physiol. 1996; Scholar, J.M. J. Clin. Invest. 1991; PubMed Scopus Google Scholar, R. J.M. J. Am. J. Physiol. 1993; Google Scholar, A. P.E. J. Clin. Invest. 1994; PubMed Scopus Google Scholar). is that these changes in calcium to the changes in biochemical of TJ proteins and cytoskeletal association which have after ATP depletion. a high calcium is to an association, at least in which the TJ is calcium is with or calcium in the absence of ATP depletion, is increase in the of TJ proteins changes in calcium an in TJ after TJ disassembly in the of ATP the a in the of these during reassembly after ATP is for a in the calcium changes in calcium have been at least to the of TJ proteins R.O. Sun A. Bush K.T. Nigam S.K. J. Biol. Chem. 1996; 271: 13636-13641Abstract PubMed Scopus Google Scholar). In this model for TJ disassembly and the reassembly of the TJ a protein B.M. C. S. Nigam S.K. J. Biol. Chem. 1996; 271: PubMed Scopus Google calcium R.O. Sun A. Panichas M. Hebert S.C. Brenner B.M. Nigam S.K. J. Cell. Physiol. 1994; 159: 423-433Crossref PubMed Scopus (105) Google Scholar, 19Stuart R.O. Sun A. Bush K.T. Nigam S.K. J. Biol. Chem. 1996; 271: 13636-13641Abstract PubMed Scopus Google and protein (8Stuart R.O. Nigam S.K. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 6072-6076Crossref PubMed Scopus (242) Google Scholar). to in studies the reassembly of the TJ after ATP is a to link a number of membrane proteins to the cytoskeleton W.J. J. Cell Biol. PubMed Scopus Google Scholar, W.J. E.M. J. Cell Biol. 1990; PubMed Scopus (244) Google Scholar, S. Tsukita S. Nagafuchi A. Yonemura S. Curr. Opin. Cell Biol. 1992; PubMed Scopus Google Scholar). as well as has been to with proteins in epithelial cells Physiol. 1990; Scopus Google Scholar, Nelson W.J. Am. J. Physiol. 1996; PubMed Google Scholar). data that after ATP depletion, the fraction of fodrin the TJ Although to fodrin with the ZO-1-containing this association is or our that the of between the TJ and cytoskeleton is in the of epithelial cell ischemic and propose that a key step in this is the association of TJ proteins with fodrin and/or other cytoskeletal proteins, an association that by mechanisms involving calcium and/or protein that with this association of in the of ischemic
Tsukamoto et al. (Sun,) studied this question.
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