JNK/c-Jun signaling mediates an anti-apoptotic effect of RANKL in osteoclasts

Fumiyo Ikeda, Takuma Matsubara, Taro Tsurukai, Kenji Hata, Riko Nishimura, Toshiyuki Yoneda

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Abstract

Introduction: RANKL is known to be important not only for differentiation and activation of osteoclasts but also for their survival. Experimentally, apoptosis of osteoclasts is rapidly induced by the deprivation of RANKL. RANKL activates Elk-related tyrosine kinase (ERK), p38, c-Jun N-terminal kinase (JNK), and NF-κB pathways through TRAF6 in osteoclasts and the precursor cells. It has been shown that ERK is critical for regulation of osteoclast survival. However, an involvement of other RANKL signaling pathways such as JNK signaling in survival of osteoclasts has not been fully understood yet. Materials and Methods: Osteoclasts derived from primary mouse bone marrow cells by soluble RANKL (sRANKL) were treated with a JNK inhibitor, SP600125, or infected with adenovirus carrying dominant-negative (DN)-c-jun, DN-c-fos, mitogen-activated protein kinase kinase 1 (MEKK1), I-κBα mutant, or NF-κB components, p50 and p65. Osteoclasts were cultured with or without sRANKL, and apoptotic phenotype was determined by TUNEL assay, DAPI staining, and expression of cleaved caspase 3 followed by TRACP staining. Results: Overexpression of TRAF6 activated JNK and NF-κB signaling pathways and clearly prevented osteoclasts from apoptosis caused by abrogation of sRANKL. An anti-apoptotic effect of RANKL/RANK/TRAF6 signaling on osteoclast was inhibited by JNK-specific inhibitor SP600125 and by overexpression of dominant-negative JNK1, c-jun, and c-fos. Also, overexpression of MEKK1 inhibited apoptosis of osteoclasts even in the absence of sRANKL along with activation of JNK/c-jun signaling. On the other hand, blockade of NF-κB signaling by I-κBα mutant or overexpression of NF-κB components showed a marginal effect on apoptosis of osteoclasts. Conclusions: An important role of RANKL-induced activation of MEKK1/JNK/c-jun signaling in the regulation of apoptosis in osteoclasts was shown. Our study suggests that c-fos plays a role as a partner of activator protein-1 factor, c-jun, during the regulation of apoptosis in osteoclasts.

Original languageEnglish
Pages (from-to)907-914
Number of pages8
JournalJournal of Bone and Mineral Research
Volume23
Issue number6
DOIs
Publication statusPublished - Jun 1 2008

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JNK Mitogen-Activated Protein Kinases
Osteoclasts
TNF Receptor-Associated Factor 6
MAP Kinase Kinase 1
Apoptosis
Protein-Tyrosine Kinases
Staining and Labeling
Transcription Factor AP-1
In Situ Nick-End Labeling
Adenoviridae
Caspase 3
Bone Marrow Cells

All Science Journal Classification (ASJC) codes

  • Endocrinology, Diabetes and Metabolism
  • Orthopedics and Sports Medicine

Cite this

JNK/c-Jun signaling mediates an anti-apoptotic effect of RANKL in osteoclasts. / Ikeda, Fumiyo; Matsubara, Takuma; Tsurukai, Taro; Hata, Kenji; Nishimura, Riko; Yoneda, Toshiyuki.

In: Journal of Bone and Mineral Research, Vol. 23, No. 6, 01.06.2008, p. 907-914.

Research output: Contribution to journalArticle

Ikeda, F, Matsubara, T, Tsurukai, T, Hata, K, Nishimura, R & Yoneda, T 2008, 'JNK/c-Jun signaling mediates an anti-apoptotic effect of RANKL in osteoclasts', Journal of Bone and Mineral Research, vol. 23, no. 6, pp. 907-914. https://doi.org/10.1359/jbmr.080211
Ikeda, Fumiyo ; Matsubara, Takuma ; Tsurukai, Taro ; Hata, Kenji ; Nishimura, Riko ; Yoneda, Toshiyuki. / JNK/c-Jun signaling mediates an anti-apoptotic effect of RANKL in osteoclasts. In: Journal of Bone and Mineral Research. 2008 ; Vol. 23, No. 6. pp. 907-914.
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abstract = "Introduction: RANKL is known to be important not only for differentiation and activation of osteoclasts but also for their survival. Experimentally, apoptosis of osteoclasts is rapidly induced by the deprivation of RANKL. RANKL activates Elk-related tyrosine kinase (ERK), p38, c-Jun N-terminal kinase (JNK), and NF-κB pathways through TRAF6 in osteoclasts and the precursor cells. It has been shown that ERK is critical for regulation of osteoclast survival. However, an involvement of other RANKL signaling pathways such as JNK signaling in survival of osteoclasts has not been fully understood yet. Materials and Methods: Osteoclasts derived from primary mouse bone marrow cells by soluble RANKL (sRANKL) were treated with a JNK inhibitor, SP600125, or infected with adenovirus carrying dominant-negative (DN)-c-jun, DN-c-fos, mitogen-activated protein kinase kinase 1 (MEKK1), I-κBα mutant, or NF-κB components, p50 and p65. Osteoclasts were cultured with or without sRANKL, and apoptotic phenotype was determined by TUNEL assay, DAPI staining, and expression of cleaved caspase 3 followed by TRACP staining. Results: Overexpression of TRAF6 activated JNK and NF-κB signaling pathways and clearly prevented osteoclasts from apoptosis caused by abrogation of sRANKL. An anti-apoptotic effect of RANKL/RANK/TRAF6 signaling on osteoclast was inhibited by JNK-specific inhibitor SP600125 and by overexpression of dominant-negative JNK1, c-jun, and c-fos. Also, overexpression of MEKK1 inhibited apoptosis of osteoclasts even in the absence of sRANKL along with activation of JNK/c-jun signaling. On the other hand, blockade of NF-κB signaling by I-κBα mutant or overexpression of NF-κB components showed a marginal effect on apoptosis of osteoclasts. Conclusions: An important role of RANKL-induced activation of MEKK1/JNK/c-jun signaling in the regulation of apoptosis in osteoclasts was shown. Our study suggests that c-fos plays a role as a partner of activator protein-1 factor, c-jun, during the regulation of apoptosis in osteoclasts.",
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T1 - JNK/c-Jun signaling mediates an anti-apoptotic effect of RANKL in osteoclasts

AU - Ikeda, Fumiyo

AU - Matsubara, Takuma

AU - Tsurukai, Taro

AU - Hata, Kenji

AU - Nishimura, Riko

AU - Yoneda, Toshiyuki

PY - 2008/6/1

Y1 - 2008/6/1

N2 - Introduction: RANKL is known to be important not only for differentiation and activation of osteoclasts but also for their survival. Experimentally, apoptosis of osteoclasts is rapidly induced by the deprivation of RANKL. RANKL activates Elk-related tyrosine kinase (ERK), p38, c-Jun N-terminal kinase (JNK), and NF-κB pathways through TRAF6 in osteoclasts and the precursor cells. It has been shown that ERK is critical for regulation of osteoclast survival. However, an involvement of other RANKL signaling pathways such as JNK signaling in survival of osteoclasts has not been fully understood yet. Materials and Methods: Osteoclasts derived from primary mouse bone marrow cells by soluble RANKL (sRANKL) were treated with a JNK inhibitor, SP600125, or infected with adenovirus carrying dominant-negative (DN)-c-jun, DN-c-fos, mitogen-activated protein kinase kinase 1 (MEKK1), I-κBα mutant, or NF-κB components, p50 and p65. Osteoclasts were cultured with or without sRANKL, and apoptotic phenotype was determined by TUNEL assay, DAPI staining, and expression of cleaved caspase 3 followed by TRACP staining. Results: Overexpression of TRAF6 activated JNK and NF-κB signaling pathways and clearly prevented osteoclasts from apoptosis caused by abrogation of sRANKL. An anti-apoptotic effect of RANKL/RANK/TRAF6 signaling on osteoclast was inhibited by JNK-specific inhibitor SP600125 and by overexpression of dominant-negative JNK1, c-jun, and c-fos. Also, overexpression of MEKK1 inhibited apoptosis of osteoclasts even in the absence of sRANKL along with activation of JNK/c-jun signaling. On the other hand, blockade of NF-κB signaling by I-κBα mutant or overexpression of NF-κB components showed a marginal effect on apoptosis of osteoclasts. Conclusions: An important role of RANKL-induced activation of MEKK1/JNK/c-jun signaling in the regulation of apoptosis in osteoclasts was shown. Our study suggests that c-fos plays a role as a partner of activator protein-1 factor, c-jun, during the regulation of apoptosis in osteoclasts.

AB - Introduction: RANKL is known to be important not only for differentiation and activation of osteoclasts but also for their survival. Experimentally, apoptosis of osteoclasts is rapidly induced by the deprivation of RANKL. RANKL activates Elk-related tyrosine kinase (ERK), p38, c-Jun N-terminal kinase (JNK), and NF-κB pathways through TRAF6 in osteoclasts and the precursor cells. It has been shown that ERK is critical for regulation of osteoclast survival. However, an involvement of other RANKL signaling pathways such as JNK signaling in survival of osteoclasts has not been fully understood yet. Materials and Methods: Osteoclasts derived from primary mouse bone marrow cells by soluble RANKL (sRANKL) were treated with a JNK inhibitor, SP600125, or infected with adenovirus carrying dominant-negative (DN)-c-jun, DN-c-fos, mitogen-activated protein kinase kinase 1 (MEKK1), I-κBα mutant, or NF-κB components, p50 and p65. Osteoclasts were cultured with or without sRANKL, and apoptotic phenotype was determined by TUNEL assay, DAPI staining, and expression of cleaved caspase 3 followed by TRACP staining. Results: Overexpression of TRAF6 activated JNK and NF-κB signaling pathways and clearly prevented osteoclasts from apoptosis caused by abrogation of sRANKL. An anti-apoptotic effect of RANKL/RANK/TRAF6 signaling on osteoclast was inhibited by JNK-specific inhibitor SP600125 and by overexpression of dominant-negative JNK1, c-jun, and c-fos. Also, overexpression of MEKK1 inhibited apoptosis of osteoclasts even in the absence of sRANKL along with activation of JNK/c-jun signaling. On the other hand, blockade of NF-κB signaling by I-κBα mutant or overexpression of NF-κB components showed a marginal effect on apoptosis of osteoclasts. Conclusions: An important role of RANKL-induced activation of MEKK1/JNK/c-jun signaling in the regulation of apoptosis in osteoclasts was shown. Our study suggests that c-fos plays a role as a partner of activator protein-1 factor, c-jun, during the regulation of apoptosis in osteoclasts.

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