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ROCK activity regulates functional tight junction assembly during blastocyst formation in porcine parthenogenetic embryos

The Rho-associated coiled-coil-containing protein serine/threonine kinases 1 and 2 (ROCK1 and ROCK2) are Rho subfamily GTPase downstream effectors that regulate cell migration, intercellular adhesion, cell polarity, and cell proliferation by stimulating actin cytoskeleton reorganization. Inhibition...

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Autores principales: Kwon, Jeongwoo, Kim, Nam-Hyung, Choi, Inchul
Formato: Online Artículo Texto
Lenguaje:English
Publicado: PeerJ Inc. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4830244/
https://www.ncbi.nlm.nih.gov/pubmed/27077008
http://dx.doi.org/10.7717/peerj.1914
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author Kwon, Jeongwoo
Kim, Nam-Hyung
Choi, Inchul
author_facet Kwon, Jeongwoo
Kim, Nam-Hyung
Choi, Inchul
author_sort Kwon, Jeongwoo
collection PubMed
description The Rho-associated coiled-coil-containing protein serine/threonine kinases 1 and 2 (ROCK1 and ROCK2) are Rho subfamily GTPase downstream effectors that regulate cell migration, intercellular adhesion, cell polarity, and cell proliferation by stimulating actin cytoskeleton reorganization. Inhibition of ROCK proteins affects specification of the trophectoderm (TE) and inner cell mass (ICM) lineages, compaction, and blastocyst cavitation. However, the molecules involved in blastocyst formation are not known. Here, we examined developmental competence and levels of adherens/tight junction (AJ/TJ) constituent proteins, such as CXADR, OCLN, TJP1, and CDH1, as well as expression of their respective mRNAs, after treating porcine parthenogenetic four-cell embryos with Y-27632, a specific inhibitor of ROCK, at concentrations of 0, 10, 20, 100 µM for 24 h. Following this treatment, the blastocyst development rates were 39.1, 20.7, 10.0, and 0% respectively. In embryos treated with 20 µM treatment, expression levels of CXADR, OCLN, TJP1, and CDH1 mRNA and protein molecules were significantly reduced (P < 0.05). FITC-dextran uptake assay revealed that the treatment caused an increase in TE TJ permeability. Interestingly, the majority of the four-cell and morula embryos treated with 20 µM Y-27643 for 24 h showed defective compaction and cavitation. Taken together, our results indicate that ROCK activity may differentially affect assembly of AJ/TJs as well as regulate expression of genes encoding junctional proteins.
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spelling pubmed-48302442016-04-13 ROCK activity regulates functional tight junction assembly during blastocyst formation in porcine parthenogenetic embryos Kwon, Jeongwoo Kim, Nam-Hyung Choi, Inchul PeerJ Cell Biology The Rho-associated coiled-coil-containing protein serine/threonine kinases 1 and 2 (ROCK1 and ROCK2) are Rho subfamily GTPase downstream effectors that regulate cell migration, intercellular adhesion, cell polarity, and cell proliferation by stimulating actin cytoskeleton reorganization. Inhibition of ROCK proteins affects specification of the trophectoderm (TE) and inner cell mass (ICM) lineages, compaction, and blastocyst cavitation. However, the molecules involved in blastocyst formation are not known. Here, we examined developmental competence and levels of adherens/tight junction (AJ/TJ) constituent proteins, such as CXADR, OCLN, TJP1, and CDH1, as well as expression of their respective mRNAs, after treating porcine parthenogenetic four-cell embryos with Y-27632, a specific inhibitor of ROCK, at concentrations of 0, 10, 20, 100 µM for 24 h. Following this treatment, the blastocyst development rates were 39.1, 20.7, 10.0, and 0% respectively. In embryos treated with 20 µM treatment, expression levels of CXADR, OCLN, TJP1, and CDH1 mRNA and protein molecules were significantly reduced (P < 0.05). FITC-dextran uptake assay revealed that the treatment caused an increase in TE TJ permeability. Interestingly, the majority of the four-cell and morula embryos treated with 20 µM Y-27643 for 24 h showed defective compaction and cavitation. Taken together, our results indicate that ROCK activity may differentially affect assembly of AJ/TJs as well as regulate expression of genes encoding junctional proteins. PeerJ Inc. 2016-04-11 /pmc/articles/PMC4830244/ /pubmed/27077008 http://dx.doi.org/10.7717/peerj.1914 Text en ©2016 Kwon et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, reproduction and adaptation in any medium and for any purpose provided that it is properly attributed. For attribution, the original author(s), title, publication source (PeerJ) and either DOI or URL of the article must be cited.
spellingShingle Cell Biology
Kwon, Jeongwoo
Kim, Nam-Hyung
Choi, Inchul
ROCK activity regulates functional tight junction assembly during blastocyst formation in porcine parthenogenetic embryos
title ROCK activity regulates functional tight junction assembly during blastocyst formation in porcine parthenogenetic embryos
title_full ROCK activity regulates functional tight junction assembly during blastocyst formation in porcine parthenogenetic embryos
title_fullStr ROCK activity regulates functional tight junction assembly during blastocyst formation in porcine parthenogenetic embryos
title_full_unstemmed ROCK activity regulates functional tight junction assembly during blastocyst formation in porcine parthenogenetic embryos
title_short ROCK activity regulates functional tight junction assembly during blastocyst formation in porcine parthenogenetic embryos
title_sort rock activity regulates functional tight junction assembly during blastocyst formation in porcine parthenogenetic embryos
topic Cell Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4830244/
https://www.ncbi.nlm.nih.gov/pubmed/27077008
http://dx.doi.org/10.7717/peerj.1914
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AT choiinchul rockactivityregulatesfunctionaltightjunctionassemblyduringblastocystformationinporcineparthenogeneticembryos