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ERK5 activation is essential for osteoclast differentiation.

Amano S, Chang YT, Fukui Y - PLoS ONE (2015)

Bottom Line: Therefore, activation of ERK5 is required for the induction of c-Fos.These events were confirmed in experiments using M-CSF-dependent bone marrow macrophages.Taken together, the present results show that activation of the MEK5/ERK5 pathway with M-CSF is required for osteoclast differentiation, which may induce differentiation through the induction of c-Fos.

View Article: PubMed Central - PubMed

Affiliation: Division of Microbiology and Immunology, Department of Oral Biology and Tissue Engineering, Meikai University School of Dentistry, Keyakidai, Sakado City, Japan.

ABSTRACT
The MEK/ERK pathways are critical for controlling cell proliferation and differentiation. In this study, we show that the MEK5/ERK5 pathway participates in osteoclast differentiation. ERK5 was activated by M-CSF, which is one of the essential factors in osteoclast differentiation. Inhibition of MEK5 by BIX02189 or inhibition of ERK5 by XMD 8-92 blocked osteoclast differentiation. MEK5 knockdown inhibited osteoclast differentiation. RAW264.7D clone cells, which are monocytic cells, differentiate into osteoclasts after stimulation with sRANKL. ERK5 was activated without any stimulation in these cells. Inhibition of the MEK5/ERK5 pathway by the inhibitors also blocked the differentiation of RAW264.7D cells into osteoclasts. Moreover, expression of the transcription factor c-Fos, which is indispensable for osteoclast differentiation, was inhibited by treatment with MEK5 or ERK5 inhibitors. Therefore, activation of ERK5 is required for the induction of c-Fos. These events were confirmed in experiments using M-CSF-dependent bone marrow macrophages. Taken together, the present results show that activation of the MEK5/ERK5 pathway with M-CSF is required for osteoclast differentiation, which may induce differentiation through the induction of c-Fos.

No MeSH data available.


Related in: MedlinePlus

The induction of c-Fos was blocked by inhibition of the ERK5 pathway.(A) 4B12 cells (2.5 × 104) were cultured with M-CSF (10 ng/ml) alone (M) or with M-CSF (10 ng/ml) and sRANKL (10 ng/ml) (MR). After the indicated times, c-Fos gene expression was measured by qRT-PCR. (B, C) 4B12 cells (2.5 × 104) were pretreated with or without BIX02189 or XMD8-92 for 6 hrs and then cultured in the presence of M-CSF (10 ng/ml) for 24 hrs. The expression of c-Fos was measured by qRT-PCR and Western blot analysis. Similar results were obtained in two independent experiments.
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pone.0125054.g005: The induction of c-Fos was blocked by inhibition of the ERK5 pathway.(A) 4B12 cells (2.5 × 104) were cultured with M-CSF (10 ng/ml) alone (M) or with M-CSF (10 ng/ml) and sRANKL (10 ng/ml) (MR). After the indicated times, c-Fos gene expression was measured by qRT-PCR. (B, C) 4B12 cells (2.5 × 104) were pretreated with or without BIX02189 or XMD8-92 for 6 hrs and then cultured in the presence of M-CSF (10 ng/ml) for 24 hrs. The expression of c-Fos was measured by qRT-PCR and Western blot analysis. Similar results were obtained in two independent experiments.

Mentions: Given that ERK5 can significantly induce c-Fos, we tested whether the induction of c-Fos in 4B12 cells was affected by ERK5 activation [8,9]. c-Fos mRNA expression was induced in 4B12 cells in response to stimulation with M-CSF alone, and this expression was enhanced following stimulation with M-CSF and sRANKL (Fig 5A). Pretreatment with both BIX02189 and XMD8-92 significantly inhibited M-CSF-induced c-Fos expression at the mRNA and protein levels (Fig 5B and 5C). These results suggest that ERK5 activation is required for the induction of c-Fos.


ERK5 activation is essential for osteoclast differentiation.

Amano S, Chang YT, Fukui Y - PLoS ONE (2015)

The induction of c-Fos was blocked by inhibition of the ERK5 pathway.(A) 4B12 cells (2.5 × 104) were cultured with M-CSF (10 ng/ml) alone (M) or with M-CSF (10 ng/ml) and sRANKL (10 ng/ml) (MR). After the indicated times, c-Fos gene expression was measured by qRT-PCR. (B, C) 4B12 cells (2.5 × 104) were pretreated with or without BIX02189 or XMD8-92 for 6 hrs and then cultured in the presence of M-CSF (10 ng/ml) for 24 hrs. The expression of c-Fos was measured by qRT-PCR and Western blot analysis. Similar results were obtained in two independent experiments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4401765&req=5

pone.0125054.g005: The induction of c-Fos was blocked by inhibition of the ERK5 pathway.(A) 4B12 cells (2.5 × 104) were cultured with M-CSF (10 ng/ml) alone (M) or with M-CSF (10 ng/ml) and sRANKL (10 ng/ml) (MR). After the indicated times, c-Fos gene expression was measured by qRT-PCR. (B, C) 4B12 cells (2.5 × 104) were pretreated with or without BIX02189 or XMD8-92 for 6 hrs and then cultured in the presence of M-CSF (10 ng/ml) for 24 hrs. The expression of c-Fos was measured by qRT-PCR and Western blot analysis. Similar results were obtained in two independent experiments.
Mentions: Given that ERK5 can significantly induce c-Fos, we tested whether the induction of c-Fos in 4B12 cells was affected by ERK5 activation [8,9]. c-Fos mRNA expression was induced in 4B12 cells in response to stimulation with M-CSF alone, and this expression was enhanced following stimulation with M-CSF and sRANKL (Fig 5A). Pretreatment with both BIX02189 and XMD8-92 significantly inhibited M-CSF-induced c-Fos expression at the mRNA and protein levels (Fig 5B and 5C). These results suggest that ERK5 activation is required for the induction of c-Fos.

Bottom Line: Therefore, activation of ERK5 is required for the induction of c-Fos.These events were confirmed in experiments using M-CSF-dependent bone marrow macrophages.Taken together, the present results show that activation of the MEK5/ERK5 pathway with M-CSF is required for osteoclast differentiation, which may induce differentiation through the induction of c-Fos.

View Article: PubMed Central - PubMed

Affiliation: Division of Microbiology and Immunology, Department of Oral Biology and Tissue Engineering, Meikai University School of Dentistry, Keyakidai, Sakado City, Japan.

ABSTRACT
The MEK/ERK pathways are critical for controlling cell proliferation and differentiation. In this study, we show that the MEK5/ERK5 pathway participates in osteoclast differentiation. ERK5 was activated by M-CSF, which is one of the essential factors in osteoclast differentiation. Inhibition of MEK5 by BIX02189 or inhibition of ERK5 by XMD 8-92 blocked osteoclast differentiation. MEK5 knockdown inhibited osteoclast differentiation. RAW264.7D clone cells, which are monocytic cells, differentiate into osteoclasts after stimulation with sRANKL. ERK5 was activated without any stimulation in these cells. Inhibition of the MEK5/ERK5 pathway by the inhibitors also blocked the differentiation of RAW264.7D cells into osteoclasts. Moreover, expression of the transcription factor c-Fos, which is indispensable for osteoclast differentiation, was inhibited by treatment with MEK5 or ERK5 inhibitors. Therefore, activation of ERK5 is required for the induction of c-Fos. These events were confirmed in experiments using M-CSF-dependent bone marrow macrophages. Taken together, the present results show that activation of the MEK5/ERK5 pathway with M-CSF is required for osteoclast differentiation, which may induce differentiation through the induction of c-Fos.

No MeSH data available.


Related in: MedlinePlus