Osteogenic gene transcription is regulated via gap junction-mediated cell-cell communication.
Osteogenic gene transcription is regulated via gap junction-mediated cell-cell communication.
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DOI：
10.1089/scd.2014.0060
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发表时间：
2015
影响因子：
4
通讯作者：
Yoshikazu Mikami;Kiyofumi Yamamoto;Yuko Akiyama;Masayuki Kobayashi;Eri Watanabe;N. Watanabe;M. Asano;N. Shimizu;K. Komiyama
中科院分区：
文献类型：
--
作者：
Yoshikazu Mikami;Kiyofumi Yamamoto;Yuko Akiyama;Masayuki Kobayashi;Eri Watanabe;N. Watanabe;M. Asano;N. Shimizu;K. Komiyama
对小鼠成骨细胞样 MLO-A5 细胞和骨髓间充质干细胞 (BMSC) 样 C3H10T1/2 细胞之间的细胞间通讯进行了分析研究。生成表达绿色荧光蛋白的 C3H10T1/2 细胞（10T-GFP 细胞），以便能够从与 MLO-A5 细胞共培养物中分离 BMSC 样细胞。几种成骨转录因子（Runx2、Osterix、Dlx5 和 Msx2）的 mRNA 表达水平在共培养和单独培养的 10T-GFP 细胞之间没有差异，但碱性磷酸酶（ALP）和骨唾液蛋白（BSP）的 mRNA 表达水平在共培养细胞中高出 300 至 400 倍。膜片钳和生物胞素转移测定揭示了共培养的 10T-GFP 和 MLO-A5 细胞之间间隙连接介导的通讯。添加间隙连接抑制剂可抑制共培养的 10T-GFP 细胞中 ALP 和 BSP mRNA 表达水平的增加。此外，共培养的 10T-GFP 细胞中的组蛋白乙酰化水平高于单独培养的 10T-GFP 细胞。这些结果表明成骨细胞和 BMSC 通过间隙连接结合，并且间隙连接介导的信号传导诱导组蛋白乙酰化，从而导致 BMSC 中编码 ALP 和 BSP 的基因转录升高。
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.