title: Osteogenic gene transcription is regulated via gap junction-mediated cell-cell communication.
abstract: An analytical study of cell-cell communications between murine osteoblast-like MLO-A5 cells and bone marrow mesenchymal stem cell (BMSC)-like C3H10T1/2 cells was performed. C3H10T1/2 cells expressing green fluorescent protein (10T-GFP cells) were generated to enable the isolation of the BMSC-like cells from co-cultures with MLO-A5 cells. The mRNA expression levels of several osteogenic transcription factors (Runx2, Osterix, Dlx5, and Msx2) did not differ between the co-cultured and mono-cultured 10T-GFP cells, but those of alkaline phosphatase (ALP) and bone sialoprotein (BSP) were 300- to 400-fold higher in the co-cultured cells. Patch clamp and biocytin transfer assays revealed gap junction-mediated communication between co-cultured 10T-GFP and MLO-A5 cells. The addition of a gap junction inhibitor suppressed the increases in the expression levels of the ALP and BSP mRNAs in co-cultured 10T-GFP cells. Furthermore, the histone acetylation levels were higher in co-cultured 10T-GFP cells than in mono-cultured 10T-GFP cells. These results suggest that osteoblasts and BMSCs associate via gap junctions, and that gap junction-mediated signaling induces histone acetylation that leads to elevated transcription of the genes encoding ALP and BSP in BMSCs.
authors.affiliation: 1 Department of Pathology, Nihon University School of Dentistry , Tokyo, Japan .
authors.forename: Yoshikazu
authors.initials: Y
authors.lastname: Mikami
authors.forename: Kiyofumi
authors.initials: K
authors.lastname: Yamamoto
authors.forename: Yuko
authors.initials: Y
authors.lastname: Akiyama
authors.forename: Masayuki
authors.initials: M
authors.lastname: Kobayashi
authors.forename: Eri
authors.initials: E
authors.lastname: Watanabe
authors.forename: Nobukazu
authors.initials: N
authors.lastname: Watanabe
authors.forename: Masatake
authors.initials: M
authors.lastname: Asano
authors.forename: Noriyoshi
authors.initials: N
authors.lastname: Shimizu
authors.forename: Kazuo
authors.initials: K
authors.lastname: Komiyama
mesh_terms: D041721:3T3-L1 Cells; D000469:Alkaline Phosphatase / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D016466:CHO Cells; D002450:Cell Communication / Q000502:physiology*; D018920:Coculture Techniques; D006224:Cricetinae; D003412:Cricetulus; D017629:Gap Junctions / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D058574:Integrin-Binding Sialoprotein / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D010012:Osteogenesis / Q000502:physiology*; D014157:Transcription Factors / Q000096:biosynthesis / Q000235:genetics; D014158:Transcription, Genetic / Q000502:physiology*
chemical_list: C546872:Ibsp protein, mouse; D058574:Integrin-Binding Sialoprotein; D014157:Transcription Factors; D000469:Alkaline Phosphatase