Osteocytes up-regulate the terminal differentiation of pre-osteoblasts via gap junctions

Yoichi Nishikawa; Yuko Akiyama; Kiyofumi Yamamoto; Masayuki Kobayashi; Eri Watanabe; Nobukazu Watanabe; Noriyoshi Shimizu; Yoshikazu Mikami; Kazuo Komiyama

doi:10.1016/j.bbrc.2014.10.128

Osteocytes up-regulate the terminal differentiation of pre-osteoblasts via gap junctions

Yoichi Nishikawa, Yuko Akiyama, Kiyofumi Yamamoto, Masayuki Kobayashi, Eri Watanabe, Nobukazu Watanabe, Noriyoshi Shimizu, Yoshikazu Mikami, Kazuo Komiyama

School of Dentistry

Research output: Contribution to journal › Article › peer-review

12 Citations (Scopus)

Abstract

We examined cell-to-cell interaction between pre-osteoblasts and osteocytes using MC3T3-E1 and MLO-Y4, respectively. First, GFP expressing MC3T3-E1 (E1-GFP) cells were generated to isolate the cells from co-culture with MLO-Y4. No changes were observed in the expression of osteogenic transcription factors Runx2, Osterix, Dlx5 and Msx2, but expression of alkaline phosphatase (ALP) and bone sialoprotein (BSP) in E1-GFP co-cultured with MLO-Y4 was 300-400-fold greater than that in mono-cultured E1-GFP. In addition, mineralized nodule formation was drastically increased in co-cultured E1-GFP cells compared to mono-cultured cells. Patch clamp assay showed the presence of gap junctions between E1-GFP and MLO-Y4. Furthermore, when the gap junction inhibitor carbenoxolone (CBX) was added to the culture, increased expression of ALP and BSP in E1-GFP co-cultured with MLO-Y4 was suppressed. These results suggest that gap junction detected between pre-osteoblasts and osteocytes plays an important role on the terminal differentiation of pre-osteoblasts.

Original language	English
Pages (from-to)	1-6
Number of pages	6
Journal	Biochemical and Biophysical Research Communications
Volume	456
Issue number	1
DOIs	https://doi.org/10.1016/j.bbrc.2014.10.128
Publication status	Published - 2 Jan 2015

Keywords

Alkaline phosphatase
Bone sialoprotein
Gap junction
Osteoblasts
Osteocytes

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10.1016/j.bbrc.2014.10.128

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title = "Osteocytes up-regulate the terminal differentiation of pre-osteoblasts via gap junctions",

abstract = "We examined cell-to-cell interaction between pre-osteoblasts and osteocytes using MC3T3-E1 and MLO-Y4, respectively. First, GFP expressing MC3T3-E1 (E1-GFP) cells were generated to isolate the cells from co-culture with MLO-Y4. No changes were observed in the expression of osteogenic transcription factors Runx2, Osterix, Dlx5 and Msx2, but expression of alkaline phosphatase (ALP) and bone sialoprotein (BSP) in E1-GFP co-cultured with MLO-Y4 was 300-400-fold greater than that in mono-cultured E1-GFP. In addition, mineralized nodule formation was drastically increased in co-cultured E1-GFP cells compared to mono-cultured cells. Patch clamp assay showed the presence of gap junctions between E1-GFP and MLO-Y4. Furthermore, when the gap junction inhibitor carbenoxolone (CBX) was added to the culture, increased expression of ALP and BSP in E1-GFP co-cultured with MLO-Y4 was suppressed. These results suggest that gap junction detected between pre-osteoblasts and osteocytes plays an important role on the terminal differentiation of pre-osteoblasts.",

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author = "Yoichi Nishikawa and Yuko Akiyama and Kiyofumi Yamamoto and Masayuki Kobayashi and Eri Watanabe and Nobukazu Watanabe and Noriyoshi Shimizu and Yoshikazu Mikami and Kazuo Komiyama",

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AU - Nishikawa, Yoichi

AU - Akiyama, Yuko

AU - Yamamoto, Kiyofumi

AU - Kobayashi, Masayuki

AU - Watanabe, Eri

AU - Watanabe, Nobukazu

AU - Shimizu, Noriyoshi

AU - Mikami, Yoshikazu

AU - Komiyama, Kazuo

PY - 2015/1/2

Y1 - 2015/1/2

N2 - We examined cell-to-cell interaction between pre-osteoblasts and osteocytes using MC3T3-E1 and MLO-Y4, respectively. First, GFP expressing MC3T3-E1 (E1-GFP) cells were generated to isolate the cells from co-culture with MLO-Y4. No changes were observed in the expression of osteogenic transcription factors Runx2, Osterix, Dlx5 and Msx2, but expression of alkaline phosphatase (ALP) and bone sialoprotein (BSP) in E1-GFP co-cultured with MLO-Y4 was 300-400-fold greater than that in mono-cultured E1-GFP. In addition, mineralized nodule formation was drastically increased in co-cultured E1-GFP cells compared to mono-cultured cells. Patch clamp assay showed the presence of gap junctions between E1-GFP and MLO-Y4. Furthermore, when the gap junction inhibitor carbenoxolone (CBX) was added to the culture, increased expression of ALP and BSP in E1-GFP co-cultured with MLO-Y4 was suppressed. These results suggest that gap junction detected between pre-osteoblasts and osteocytes plays an important role on the terminal differentiation of pre-osteoblasts.

AB - We examined cell-to-cell interaction between pre-osteoblasts and osteocytes using MC3T3-E1 and MLO-Y4, respectively. First, GFP expressing MC3T3-E1 (E1-GFP) cells were generated to isolate the cells from co-culture with MLO-Y4. No changes were observed in the expression of osteogenic transcription factors Runx2, Osterix, Dlx5 and Msx2, but expression of alkaline phosphatase (ALP) and bone sialoprotein (BSP) in E1-GFP co-cultured with MLO-Y4 was 300-400-fold greater than that in mono-cultured E1-GFP. In addition, mineralized nodule formation was drastically increased in co-cultured E1-GFP cells compared to mono-cultured cells. Patch clamp assay showed the presence of gap junctions between E1-GFP and MLO-Y4. Furthermore, when the gap junction inhibitor carbenoxolone (CBX) was added to the culture, increased expression of ALP and BSP in E1-GFP co-cultured with MLO-Y4 was suppressed. These results suggest that gap junction detected between pre-osteoblasts and osteocytes plays an important role on the terminal differentiation of pre-osteoblasts.

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Osteocytes up-regulate the terminal differentiation of pre-osteoblasts via gap junctions

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