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Effects of Extracellular Phosphate on Gene Expression in Murine Osteoblasts

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Abstract

That phosphate homeostasis is tightly linked to skeletal mineralization is probably best underscored by the fact that the phosphaturic hormone FGF23 is primarily expressed by terminally differentiated osteoblasts/osteocytes and that increased circulating FGF23 levels are causative for different types of hypophosphatemic rickets. In contrast, FGF23 inactivation results in hyperphosphatemia, and unexpectedly this phenotype is associated with severe osteomalacia in Fgf23-deficient mice. In this context it is interesting that different cell types have been shown to respond to extracellular phosphate, thereby raising the concept that phosphate can act as a signaling molecule. To identify phosphate-responsive genes in primary murine osteoblasts we performed genome wide expression analysis with cells maintained in medium containing either 1 or 4 mM sodium phosphate for 6 h. As confirmed by qRT-PCR, this analysis revealed that several known osteoblast differentiation markers (Bglap, Ibsp, and Phex) were unaffected by raising extracellular phosphate levels. In contrast, we found that the expression of Enpp1 and Ank, two genes encoding inhibitors of matrix mineralization, was induced by extracellular phosphate, while the expression of Sost and Dkk1, two genes encoding inhibitors of bone formation, was negatively regulated. The ability of osteoblasts to respond to extracellular phosphate was dependent on their differentiation state, and shRNA-dependent repression of the phosphate transporter Slc20a1 in MC3T3-E1 cells partially abolished their molecular response to phosphate. Taken together, our results provide further evidence for a role of extracellular phosphate as a signaling molecule and raise the possibility that severe hyperphosphatemia can negatively affect skeletal mineralization.

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Acknowledgments

This work was supported by the German Federal Ministry of Education and Research within the framework of the project Molecular Pathology of Osteoporosis (OsteoPath).

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Authors and Affiliations

  1. Department of Osteology and Biomechanics, University Medical Center Hamburg Eppendorf, 20246, Hamburg, Germany

    C. Rendenbach, T. A. Yorgan, T. Heckt, C. Baldauf, A. Jeschke, J. P. David, M. Amling & T. Schinke

  2. Department of Clinical Chemistry, University Medical Center Hamburg Eppendorf, 20246, Hamburg, Germany

    B. Otto & T. Streichert

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  1. C. Rendenbach
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  2. T. A. Yorgan
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  3. T. Heckt
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Correspondence to T. Schinke.

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C. Rendenbach, T.A. Yorgan and T. Heckt have contributed equally to this work.

The authors have stated that they have no conflict of interest.

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Rendenbach, C., Yorgan, T.A., Heckt, T. et al. Effects of Extracellular Phosphate on Gene Expression in Murine Osteoblasts. Calcif Tissue Int 94, 474–483 (2014). https://doi.org/10.1007/s00223-013-9831-6

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