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Inorganic Phosphate Induces Mammalian Growth Plate Chondrocyte Apoptosis in a Mitochondrial Pathway Involving Nitric Oxide and JNK MAP Kinase

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Abstract

Chondrocytes in the hypertrophic zone of the growth plate undergo apoptosis during endochondral bone development via mechanisms that involve inorganic phosphate (Pi) and nitric oxide (NO). Recent evidence suggests that Pi-dependent NO production plays a role in apoptosis of cells in the resting zone as well. This study examined the mechanism by which Pi induces NO production and the signaling pathways by which NO mediates its effects on apoptosis in these cells. Pi decreased the number of viable cells based on MTT activity; the number of TUNEL-positive cells and the level of DNA fragmentation were increased, indicating an increase in apoptosis. Blocking NO production using the NO synthase (NOS) inhibitor l-NAME or cells from eNOS−/− mice blocked Pi-induced chondrocyte apoptosis, indicating that NO production is necessary. NO donors NOC-18 and SNOG both induced chondrocyte apoptosis. SNOG also upregulated p53 expression, the Bax/Bcl-2 expression ratio, and cytochrome c release from mitochondria, as well as caspase-3 activity, indicating that NO induces apoptosis via a mitochondrial pathway. Inhibition of JNK, but not of p38 or ERK1/2, MAP kinase was able to block NO-induced apoptosis, indicating that JNK is necessary in this pathway. Pi elevates NO production via eNOS in resting zone chondrocytes, which leads to a mitochondrial apoptosis pathway dependent on JNK.

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Acknowledgments

This research was funded by a grant from Children’s Healthcare of Atlanta. The authors thank Ms. Sharon Hyzy for her assistance in preparing the manuscript.

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Authors and Affiliations

  1. Wallace H. Coulter Department of Biomedical Engineering at Georgia Tech and Emory University, Georgia Institute of Technology, 315 Ferst Drive NW, Atlanta, GA, 30332-0363, USA

    M. Zhong, H. Jo, B. D. Boyan & Z. Schwartz

  2. University of Texas Medical Branch, Galveston, TX, USA

    D. H. Carney

  3. Emory University Medical School, Atlanta, GA, USA

    H. Jo

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  1. M. Zhong
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  4. B. D. Boyan
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Correspondence to B. D. Boyan.

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Zhong, M., Carney, D.H., Jo, H. et al. Inorganic Phosphate Induces Mammalian Growth Plate Chondrocyte Apoptosis in a Mitochondrial Pathway Involving Nitric Oxide and JNK MAP Kinase. Calcif Tissue Int 88, 96–108 (2011). https://doi.org/10.1007/s00223-010-9433-5

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  • DOI: https://doi.org/10.1007/s00223-010-9433-5

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