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Connexin 43 Hemichannels Regulate Osteoblast to Osteocyte Differentiation

Connexin 43 (Cx43) is the predominant connexin subtype expressed in osteocytes. Osteocytes, accounting for 90%–95% of total bone cells, function as orchestrators coordinating balanced activity between bone-resorbing osteoclasts and bone-forming osteoblasts. In this study, two newly developed osteocy...

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Autores principales: Hua, Rui, Gu, Sumin, Jiang, Jean X.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9184820/
https://www.ncbi.nlm.nih.gov/pubmed/35693933
http://dx.doi.org/10.3389/fcell.2022.892229
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author Hua, Rui
Gu, Sumin
Jiang, Jean X.
author_facet Hua, Rui
Gu, Sumin
Jiang, Jean X.
author_sort Hua, Rui
collection PubMed
description Connexin 43 (Cx43) is the predominant connexin subtype expressed in osteocytes. Osteocytes, accounting for 90%–95% of total bone cells, function as orchestrators coordinating balanced activity between bone-resorbing osteoclasts and bone-forming osteoblasts. In this study, two newly developed osteocytic cell lines, OCY454 and IDG-SW3, were used to determine the role of Cx43 gap junctions and hemichannels (HCs) in the regulation of osteoblast to osteocyte differentiation. We found that the Cx43 level was substantially increased during the differentiation of IDG-SW3 cells and is also much higher than that of OCY454 cells. We knocked down Cx43 expression using the lentiviral CRISPR/Cas9 approach and inhibition of Cx43 HCs using Cx43 (E2) antibody in IDG-SW3 cells. Cx43 knockdown (KD) or Cx43 HC inhibition decreased gene expression for osteoblast and osteocyte markers, including alkaline phosphatase, type I collagen, dentin matrix protein 1, sclerostin, and fibroblast growth factor 23, whereas increasing the osteoclastogenesis indicator and the receptor activator of nuclear factor kappa-B ligand (RANKL)/osteoprotegerin (OPG) ratio at early and late differentiation stages. Moreover, mineralization was remarkably attenuated in differentiated Cx43-deficient IDG-SW3 cells compared to ROSA26 control. The conditioned medium collected from fully differentiated IDG-SW3 cells with Cx43 KD promoted osteoclastogenesis of RAW264.7 osteoclast precursors. Our results demonstrated that Cx43 HCs play critical roles in osteoblast to osteocyte differentiation process and regulate osteoclast differentiation via secreted factors.
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spelling pubmed-91848202022-06-11 Connexin 43 Hemichannels Regulate Osteoblast to Osteocyte Differentiation Hua, Rui Gu, Sumin Jiang, Jean X. Front Cell Dev Biol Cell and Developmental Biology Connexin 43 (Cx43) is the predominant connexin subtype expressed in osteocytes. Osteocytes, accounting for 90%–95% of total bone cells, function as orchestrators coordinating balanced activity between bone-resorbing osteoclasts and bone-forming osteoblasts. In this study, two newly developed osteocytic cell lines, OCY454 and IDG-SW3, were used to determine the role of Cx43 gap junctions and hemichannels (HCs) in the regulation of osteoblast to osteocyte differentiation. We found that the Cx43 level was substantially increased during the differentiation of IDG-SW3 cells and is also much higher than that of OCY454 cells. We knocked down Cx43 expression using the lentiviral CRISPR/Cas9 approach and inhibition of Cx43 HCs using Cx43 (E2) antibody in IDG-SW3 cells. Cx43 knockdown (KD) or Cx43 HC inhibition decreased gene expression for osteoblast and osteocyte markers, including alkaline phosphatase, type I collagen, dentin matrix protein 1, sclerostin, and fibroblast growth factor 23, whereas increasing the osteoclastogenesis indicator and the receptor activator of nuclear factor kappa-B ligand (RANKL)/osteoprotegerin (OPG) ratio at early and late differentiation stages. Moreover, mineralization was remarkably attenuated in differentiated Cx43-deficient IDG-SW3 cells compared to ROSA26 control. The conditioned medium collected from fully differentiated IDG-SW3 cells with Cx43 KD promoted osteoclastogenesis of RAW264.7 osteoclast precursors. Our results demonstrated that Cx43 HCs play critical roles in osteoblast to osteocyte differentiation process and regulate osteoclast differentiation via secreted factors. Frontiers Media S.A. 2022-05-27 /pmc/articles/PMC9184820/ /pubmed/35693933 http://dx.doi.org/10.3389/fcell.2022.892229 Text en Copyright © 2022 Hua, Gu and Jiang. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Hua, Rui
Gu, Sumin
Jiang, Jean X.
Connexin 43 Hemichannels Regulate Osteoblast to Osteocyte Differentiation
title Connexin 43 Hemichannels Regulate Osteoblast to Osteocyte Differentiation
title_full Connexin 43 Hemichannels Regulate Osteoblast to Osteocyte Differentiation
title_fullStr Connexin 43 Hemichannels Regulate Osteoblast to Osteocyte Differentiation
title_full_unstemmed Connexin 43 Hemichannels Regulate Osteoblast to Osteocyte Differentiation
title_short Connexin 43 Hemichannels Regulate Osteoblast to Osteocyte Differentiation
title_sort connexin 43 hemichannels regulate osteoblast to osteocyte differentiation
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9184820/
https://www.ncbi.nlm.nih.gov/pubmed/35693933
http://dx.doi.org/10.3389/fcell.2022.892229
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