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Molecular profiling of the basement membrane of pluripotent epiblast cells in post-implantation stage mouse embryos

INTRODUCTION: The basement membrane (BM) is a sheet-like extracellular matrix (ECM) lining the basal side of epithelial and endothelial cells. The molecular composition of the BM diversifies as embryonic development proceeds, providing optimized microenvironments for individual cell types. In post-i...

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Autores principales: Futaki, Sugiko, Nakano, Itsuko, Kawasaki, Miwa, Sanzen, Noriko, Sekiguchi, Kiyotoshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Japanese Society for Regenerative Medicine 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6933449/
https://www.ncbi.nlm.nih.gov/pubmed/31890767
http://dx.doi.org/10.1016/j.reth.2019.04.010
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author Futaki, Sugiko
Nakano, Itsuko
Kawasaki, Miwa
Sanzen, Noriko
Sekiguchi, Kiyotoshi
author_facet Futaki, Sugiko
Nakano, Itsuko
Kawasaki, Miwa
Sanzen, Noriko
Sekiguchi, Kiyotoshi
author_sort Futaki, Sugiko
collection PubMed
description INTRODUCTION: The basement membrane (BM) is a sheet-like extracellular matrix (ECM) lining the basal side of epithelial and endothelial cells. The molecular composition of the BM diversifies as embryonic development proceeds, providing optimized microenvironments for individual cell types. In post-implantation stage embryos, the embryonic BMs are essential for differentiation of the epiblast, a layer of multipotent embryonic stem cells, and subsequent embryogenesis. To better understand the role of BMs and cell–BM interactions in early embryogenesis, it is imperative to accumulate information on the molecular entities of the embryonic BMs. METHODS: We analyzed the expressions and localizations of 20 major BM proteins (11 laminin subunits, 6 type IV collagen subunits, nidogen-1 and -2, and perlecan) and other ECM-related proteins such as fibronectin and integrins in post-implantation stage embryos by immunohistochemistry. RESULTS: We found that a set of BM proteins, laminin α5, β1, and γ1 (comprising laminin-511), type IV collagen α1 and α2 (yielding type IV collagen α1(2)α2 [IV]), nidogen-1 and -2, and perlecan, were consistently present in the epiblast/ectoderm BMs throughout the early post-implantation stages. In contrast, laminin α1 was detected in the epiblast BM at E5.5 but decreased in later stages, suggesting that laminin-511 is a major laminin isoform in the early embryonic BM. In addition, fibronectin, a mesenchymal ECM protein, was enriched in the endoderm BM, indicating that the BM compositions differ between the ectoderm and the endoderm. Consistent with these observations, integrin α5, a high-affinity receptor for fibronectin, was localized in the endoderm, while integrin α6, a receptor for laminin-511, was localized in the ectoderm. CONCLUSIONS: The embryonic BMs underlying the epiblast/ectoderm contain a common toolkit comprising laminin-511, type IV collagen (α1(2)α2 [IV]), nidogen-1 and -2, and perlecan, providing a physiological basis for the utility of laminin-511 as a culture substrate for pluripotent stem cells. The distinctive association of laminin-511 and fibronectin with endodermal and ectodermal cells, together with the differential expression of integrin α5 and α6 in these cells, suggests that the ectodermal and endodermal cells rely on their integrin-dependent interactions with laminin-511 and fibronectin, respectively, to ensure their fate specification in embryonic development.
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spelling pubmed-69334492019-12-30 Molecular profiling of the basement membrane of pluripotent epiblast cells in post-implantation stage mouse embryos Futaki, Sugiko Nakano, Itsuko Kawasaki, Miwa Sanzen, Noriko Sekiguchi, Kiyotoshi Regen Ther Article INTRODUCTION: The basement membrane (BM) is a sheet-like extracellular matrix (ECM) lining the basal side of epithelial and endothelial cells. The molecular composition of the BM diversifies as embryonic development proceeds, providing optimized microenvironments for individual cell types. In post-implantation stage embryos, the embryonic BMs are essential for differentiation of the epiblast, a layer of multipotent embryonic stem cells, and subsequent embryogenesis. To better understand the role of BMs and cell–BM interactions in early embryogenesis, it is imperative to accumulate information on the molecular entities of the embryonic BMs. METHODS: We analyzed the expressions and localizations of 20 major BM proteins (11 laminin subunits, 6 type IV collagen subunits, nidogen-1 and -2, and perlecan) and other ECM-related proteins such as fibronectin and integrins in post-implantation stage embryos by immunohistochemistry. RESULTS: We found that a set of BM proteins, laminin α5, β1, and γ1 (comprising laminin-511), type IV collagen α1 and α2 (yielding type IV collagen α1(2)α2 [IV]), nidogen-1 and -2, and perlecan, were consistently present in the epiblast/ectoderm BMs throughout the early post-implantation stages. In contrast, laminin α1 was detected in the epiblast BM at E5.5 but decreased in later stages, suggesting that laminin-511 is a major laminin isoform in the early embryonic BM. In addition, fibronectin, a mesenchymal ECM protein, was enriched in the endoderm BM, indicating that the BM compositions differ between the ectoderm and the endoderm. Consistent with these observations, integrin α5, a high-affinity receptor for fibronectin, was localized in the endoderm, while integrin α6, a receptor for laminin-511, was localized in the ectoderm. CONCLUSIONS: The embryonic BMs underlying the epiblast/ectoderm contain a common toolkit comprising laminin-511, type IV collagen (α1(2)α2 [IV]), nidogen-1 and -2, and perlecan, providing a physiological basis for the utility of laminin-511 as a culture substrate for pluripotent stem cells. The distinctive association of laminin-511 and fibronectin with endodermal and ectodermal cells, together with the differential expression of integrin α5 and α6 in these cells, suggests that the ectodermal and endodermal cells rely on their integrin-dependent interactions with laminin-511 and fibronectin, respectively, to ensure their fate specification in embryonic development. Japanese Society for Regenerative Medicine 2019-05-10 /pmc/articles/PMC6933449/ /pubmed/31890767 http://dx.doi.org/10.1016/j.reth.2019.04.010 Text en © 2019 The Japanese Society for Regenerative Medicine. Production and hosting by Elsevier B.V. http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Article
Futaki, Sugiko
Nakano, Itsuko
Kawasaki, Miwa
Sanzen, Noriko
Sekiguchi, Kiyotoshi
Molecular profiling of the basement membrane of pluripotent epiblast cells in post-implantation stage mouse embryos
title Molecular profiling of the basement membrane of pluripotent epiblast cells in post-implantation stage mouse embryos
title_full Molecular profiling of the basement membrane of pluripotent epiblast cells in post-implantation stage mouse embryos
title_fullStr Molecular profiling of the basement membrane of pluripotent epiblast cells in post-implantation stage mouse embryos
title_full_unstemmed Molecular profiling of the basement membrane of pluripotent epiblast cells in post-implantation stage mouse embryos
title_short Molecular profiling of the basement membrane of pluripotent epiblast cells in post-implantation stage mouse embryos
title_sort molecular profiling of the basement membrane of pluripotent epiblast cells in post-implantation stage mouse embryos
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6933449/
https://www.ncbi.nlm.nih.gov/pubmed/31890767
http://dx.doi.org/10.1016/j.reth.2019.04.010
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