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Occludin-deficient Embryonic Stem Cells Can Differentiate into Polarized Epithelial Cells Bearing Tight Junctions

Occludin is the only known integral membrane protein of tight junctions (TJs), and is now believed to be directly involved in the barrier and fence functions of TJs. Occludin-deficient embryonic stem (ES) cells were generated by targeted disruption of both alleles of the occludin gene. When these ce...

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Autores principales: Saitou, Mitinori, Fujimoto, Kazushi, Doi, Yoshinori, Itoh, Masahiko, Fujimoto, Toyoshi, Furuse, Mikio, Takano, Hiroshi, Noda, Tetsuo, Tsukita, Shoichiro
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 1998
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2148457/
https://www.ncbi.nlm.nih.gov/pubmed/9548718
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author Saitou, Mitinori
Fujimoto, Kazushi
Doi, Yoshinori
Itoh, Masahiko
Fujimoto, Toyoshi
Furuse, Mikio
Takano, Hiroshi
Noda, Tetsuo
Tsukita, Shoichiro
author_facet Saitou, Mitinori
Fujimoto, Kazushi
Doi, Yoshinori
Itoh, Masahiko
Fujimoto, Toyoshi
Furuse, Mikio
Takano, Hiroshi
Noda, Tetsuo
Tsukita, Shoichiro
author_sort Saitou, Mitinori
collection PubMed
description Occludin is the only known integral membrane protein of tight junctions (TJs), and is now believed to be directly involved in the barrier and fence functions of TJs. Occludin-deficient embryonic stem (ES) cells were generated by targeted disruption of both alleles of the occludin gene. When these cells were subjected to suspension culture, they aggregated to form simple, and then cystic embryoid bodies (EBs) with the same time course as EB formation from wild-type ES cells. Immunofluorescence microscopy and ultrathin section electron microscopy revealed that polarized epithelial (visceral endoderm-like) cells were differentiated to delineate EBs not only from wild-type but also from occludin-deficient ES cells. Freeze fracture analyses indicated no significant differences in number or morphology of TJ strands between wild-type and occludin-deficient epithelial cells. Furthermore, zonula occludens (ZO)-1, a TJ-associated peripheral membrane protein, was still exclusively concentrated at TJ in occludin-deficient epithelial cells. In good agreement with these morphological observations, TJ in occludin-deficient epithelial cells functioned as a primary barrier to the diffusion of a low molecular mass tracer through the paracellular pathway. These findings indicate that there are as yet unidentified TJ integral membrane protein(s) which can form strand structures, recruit ZO-1, and function as a barrier without occludin.
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spelling pubmed-21484572008-05-01 Occludin-deficient Embryonic Stem Cells Can Differentiate into Polarized Epithelial Cells Bearing Tight Junctions Saitou, Mitinori Fujimoto, Kazushi Doi, Yoshinori Itoh, Masahiko Fujimoto, Toyoshi Furuse, Mikio Takano, Hiroshi Noda, Tetsuo Tsukita, Shoichiro J Cell Biol Articles Occludin is the only known integral membrane protein of tight junctions (TJs), and is now believed to be directly involved in the barrier and fence functions of TJs. Occludin-deficient embryonic stem (ES) cells were generated by targeted disruption of both alleles of the occludin gene. When these cells were subjected to suspension culture, they aggregated to form simple, and then cystic embryoid bodies (EBs) with the same time course as EB formation from wild-type ES cells. Immunofluorescence microscopy and ultrathin section electron microscopy revealed that polarized epithelial (visceral endoderm-like) cells were differentiated to delineate EBs not only from wild-type but also from occludin-deficient ES cells. Freeze fracture analyses indicated no significant differences in number or morphology of TJ strands between wild-type and occludin-deficient epithelial cells. Furthermore, zonula occludens (ZO)-1, a TJ-associated peripheral membrane protein, was still exclusively concentrated at TJ in occludin-deficient epithelial cells. In good agreement with these morphological observations, TJ in occludin-deficient epithelial cells functioned as a primary barrier to the diffusion of a low molecular mass tracer through the paracellular pathway. These findings indicate that there are as yet unidentified TJ integral membrane protein(s) which can form strand structures, recruit ZO-1, and function as a barrier without occludin. The Rockefeller University Press 1998-04-20 /pmc/articles/PMC2148457/ /pubmed/9548718 Text en This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Articles
Saitou, Mitinori
Fujimoto, Kazushi
Doi, Yoshinori
Itoh, Masahiko
Fujimoto, Toyoshi
Furuse, Mikio
Takano, Hiroshi
Noda, Tetsuo
Tsukita, Shoichiro
Occludin-deficient Embryonic Stem Cells Can Differentiate into Polarized Epithelial Cells Bearing Tight Junctions
title Occludin-deficient Embryonic Stem Cells Can Differentiate into Polarized Epithelial Cells Bearing Tight Junctions
title_full Occludin-deficient Embryonic Stem Cells Can Differentiate into Polarized Epithelial Cells Bearing Tight Junctions
title_fullStr Occludin-deficient Embryonic Stem Cells Can Differentiate into Polarized Epithelial Cells Bearing Tight Junctions
title_full_unstemmed Occludin-deficient Embryonic Stem Cells Can Differentiate into Polarized Epithelial Cells Bearing Tight Junctions
title_short Occludin-deficient Embryonic Stem Cells Can Differentiate into Polarized Epithelial Cells Bearing Tight Junctions
title_sort occludin-deficient embryonic stem cells can differentiate into polarized epithelial cells bearing tight junctions
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2148457/
https://www.ncbi.nlm.nih.gov/pubmed/9548718
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