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Comparative Analysis of Cx31 and Cx43 in Differentiation-Competent Rodent Keratinocytes

When considering connexin expression and regulation, the epidermis of the skin is one of the most complex tissues found in mammals even though it largely contains a single cell type, the keratinocyte. In the rodent epidermis, up to 9 connexin family members have been detected at the mRNA level. Many...

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Autores principales: Au, Akina, Shao, Qing, White, Kyra K., Lucaciu, Sergiu A., Esseltine, Jessica L., Barr, Kevin, Laird, Dale W.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7602205/
https://www.ncbi.nlm.nih.gov/pubmed/33066499
http://dx.doi.org/10.3390/biom10101443
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author Au, Akina
Shao, Qing
White, Kyra K.
Lucaciu, Sergiu A.
Esseltine, Jessica L.
Barr, Kevin
Laird, Dale W.
author_facet Au, Akina
Shao, Qing
White, Kyra K.
Lucaciu, Sergiu A.
Esseltine, Jessica L.
Barr, Kevin
Laird, Dale W.
author_sort Au, Akina
collection PubMed
description When considering connexin expression and regulation, the epidermis of the skin is one of the most complex tissues found in mammals even though it largely contains a single cell type, the keratinocyte. In the rodent epidermis, up to 9 connexin family members have been detected at the mRNA level. Many of these connexins are temporally and spatially regulated in coordination with keratinocyte progenitor cell differentiation and migration from the stratum basale to form the stratum spinosum and stratum granulosum layers before finally forming the stratum corneum. Cx43 is the principal connexin found in basal keratinocytes and to a lesser degree found in keratinocytes that have begun to differentiate where Cx26, Cx30 and Cx31 become prevalent. Here we show that the CRISPR-Cas9 ablation of Cx43 reduces overall gap junction coupling in monolayer cultures of rat epidermal keratinocytes (REKs) and dysregulates the differentiation of REKs when grown in organotypic cultures. Natively found in differentiated keratinocytes, Cx31 readily assembles into gap junctions when expressed in REKs where it can extensively co-assemble into the same gap junctions with co-expressed Cx30. Time-lapse imaging indicated that many Cx31 gap junctions are mobile within the plasma membrane undergoing both fusion and fission events. Finally, the persistence of pre-existing Cx31 gap junctions in the presence of the protein trafficking blocker, brefeldin A, is longer than that found for Cx43 gap junctions indicating that it has a distinctly different life expectancy in REKs. Collectively, this study highlights the importance of Cx43 in rodent keratinocyte differentiation and suggests that Cx31 acquires life-cycle properties that are distinct from Cx43.
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spelling pubmed-76022052020-11-01 Comparative Analysis of Cx31 and Cx43 in Differentiation-Competent Rodent Keratinocytes Au, Akina Shao, Qing White, Kyra K. Lucaciu, Sergiu A. Esseltine, Jessica L. Barr, Kevin Laird, Dale W. Biomolecules Article When considering connexin expression and regulation, the epidermis of the skin is one of the most complex tissues found in mammals even though it largely contains a single cell type, the keratinocyte. In the rodent epidermis, up to 9 connexin family members have been detected at the mRNA level. Many of these connexins are temporally and spatially regulated in coordination with keratinocyte progenitor cell differentiation and migration from the stratum basale to form the stratum spinosum and stratum granulosum layers before finally forming the stratum corneum. Cx43 is the principal connexin found in basal keratinocytes and to a lesser degree found in keratinocytes that have begun to differentiate where Cx26, Cx30 and Cx31 become prevalent. Here we show that the CRISPR-Cas9 ablation of Cx43 reduces overall gap junction coupling in monolayer cultures of rat epidermal keratinocytes (REKs) and dysregulates the differentiation of REKs when grown in organotypic cultures. Natively found in differentiated keratinocytes, Cx31 readily assembles into gap junctions when expressed in REKs where it can extensively co-assemble into the same gap junctions with co-expressed Cx30. Time-lapse imaging indicated that many Cx31 gap junctions are mobile within the plasma membrane undergoing both fusion and fission events. Finally, the persistence of pre-existing Cx31 gap junctions in the presence of the protein trafficking blocker, brefeldin A, is longer than that found for Cx43 gap junctions indicating that it has a distinctly different life expectancy in REKs. Collectively, this study highlights the importance of Cx43 in rodent keratinocyte differentiation and suggests that Cx31 acquires life-cycle properties that are distinct from Cx43. MDPI 2020-10-14 /pmc/articles/PMC7602205/ /pubmed/33066499 http://dx.doi.org/10.3390/biom10101443 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Au, Akina
Shao, Qing
White, Kyra K.
Lucaciu, Sergiu A.
Esseltine, Jessica L.
Barr, Kevin
Laird, Dale W.
Comparative Analysis of Cx31 and Cx43 in Differentiation-Competent Rodent Keratinocytes
title Comparative Analysis of Cx31 and Cx43 in Differentiation-Competent Rodent Keratinocytes
title_full Comparative Analysis of Cx31 and Cx43 in Differentiation-Competent Rodent Keratinocytes
title_fullStr Comparative Analysis of Cx31 and Cx43 in Differentiation-Competent Rodent Keratinocytes
title_full_unstemmed Comparative Analysis of Cx31 and Cx43 in Differentiation-Competent Rodent Keratinocytes
title_short Comparative Analysis of Cx31 and Cx43 in Differentiation-Competent Rodent Keratinocytes
title_sort comparative analysis of cx31 and cx43 in differentiation-competent rodent keratinocytes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7602205/
https://www.ncbi.nlm.nih.gov/pubmed/33066499
http://dx.doi.org/10.3390/biom10101443
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