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Altered osteoblast gluconeogenesis in X-linked hypophosphatemic mice is associated with a depressed intracellular pH

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Abstract

We have studied gluconeogenesis and intracellular pH levels in normal (+/Y) and X-linked hypophosphatemic (Hyp/Y) mice. Compared with +/Y littermates, Hyp/Y mouse osteoblasts showed a higher rate of glucose production from fructose (10-fold), glutamine, and malate, but no significant difference when α-ketoglutarate was used as substrate. The activities of the pentose cycle enzymes, glucose-6-phosphate dehydrogenase, and 6-phosphogluconate dehydrogenase, were not different in the two osteoblast preparations. Examination of intracellular pH (pHi) using the double excitation of the pH-sensitive dye 2′,7′-bis(2-carboxyethyl)-5(6)-carboxyfluorescein acetoxymethyl ester (BCECF-AM) revealed a significantly lower pHi in Hyp/Y mouse osteoblasts compared with +/Y mouse osteoblasts (7.01±0.03 n=10 versus 7.15±0.04 n=8, respectively; P<0.05). These results show for the first time that osteoblasts are capable of glucose production and that glucose production is altered in the Hyp/Y mouse osteoblast. As altered gluconeogenesis has been associated with reduced intracellular pH in other systems, a similar mechanism may be operative in the Hyp/Y mouse osteoblast. The observed defects may be intrinsic to the Hyp phenotype as the alterations in intracellular pH and gluconeogenesis persisted in vitro, or they may represent impressed memory from the in vivo state and the presumed circulating factor that influences phosphate transport.

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References

  1. Eicher EM, Southard JL, Scriver CR, Glorieux FH (1976) Hypophosphatemia: mouse model for human familial hypophosphatemic (vitamin D-resistant) rickets. Proc Natl Acad Sci USA 73:4667–4671

    Google Scholar 

  2. Meyer RA Jr, Jowsey J, Meyer MH (1979) Osteomalacia and altered magnesium metabolism in the X-linked hypophosphatemic mouse. Calcif Tissue Int 27:19–26

    Google Scholar 

  3. Meyer RA Jr, Gray RW, and Meyer MH (1980) Abnormal vitamin D metabolism in the X-linked hypophosphatemic mouse. Endocrinology 107:1577–1581

    Google Scholar 

  4. Bell CL, Tenenhouse HS, Scriver SC (1988) Primary cultures of renal epithelial cells from X-linked hypophosphatemic (Hyp) mice express defects in phosphate transport and vitamin D metabolism. Am J Hum Genet 43:293–303

    Google Scholar 

  5. Cowgill LD, Goldfarb S, Lau K, Slatopolsky E, Agus ZS (1979) Evidence for an intrinsic renal tubular defect in mice with genetic hypophosphatemic rickets. J Clin Invest 63:1203–1210

    Google Scholar 

  6. Harvey N, Tenenhouse HS (1992) Renal Na(+)-phosphate cotransport in X-linked Hyp mice responds appropriately to Na+ gradient, membrane potential, and pH. J Bone Miner Res 7:563–571

    Google Scholar 

  7. Boneh A and Tenenhouse HS (1990) Protein kinase C in mouse kidney: effect of the Hyp mutation and phosphate deprivation. Kidney Int 37:682–688

    Google Scholar 

  8. Tenenhouse HS, Henry HL (1985) Protein kinase activity and protein kinase inhibitor in mouse kidney: effect of the X-linked Hyp mutation and vitamin D status. Endocrinology 117:1719–1726

    Google Scholar 

  9. Capparelli A, Roh WD, Dhiman JK, Jo OD, Yanagawa N (1992) Altered proximal tubule glucose metabolism in X-linked hypophosphatemic mice. Endocrinology 130:328–334

    Google Scholar 

  10. Ecarot-Charrier B, Glorieux FH, Travers R, Desbarats M, Bouchard F, Hinek A (1988) Defective bone formation by transplanted Hyp mouse bone cells into normal mice. Endocrinology 123:768–773

    Google Scholar 

  11. Ecarot B, Glorieux FH, Desbarats M, Travers R, Labelle L (1992) Defective bone formation by Hyp mouse bone cells transplanted into normal mice: evidence in favor of an intrinsic osteoblast defect. J Bone Miner Res 7:215–220

    Google Scholar 

  12. Ecarot B, Glorieux FH, Desbarats M, Travers R, Labelle L (1992) Effect of dietary phosphate deprivation and supplementation of recipient mice on bone formation by transplanted cells from normal and X-linked hypophosphatemic mice. J Bone Miner Res 7:523–530

    Google Scholar 

  13. Lobaugh B, Drezner MK (1983) Abnormal regulation of renal 25-hydroxyvitamin D-1 alpha-hydroxylase activity in the X-linked hypophosphatemic mouse. J Clin Invest 71:400–403

    Google Scholar 

  14. Marie PJ, Glorieux FH (1983) Relation between hypomineralized periosteocytic lesions and bone mineralization in vitamin D-resistant rickets. Calcif Tissue Int 35:443–448

    Google Scholar 

  15. Ecarot B, Caverzasio J, Desbarats M, Bonjour JP, Glorieux FH (1994) Phosphate transport by osteoblasts from X-linked hypophosphatemic mice. Am J Physiol 266:E33–38

    Google Scholar 

  16. Nakagawa N, Arab N, Ghishan FK (1991) Characterization of the defect in the Na(+)-phosphate transporter in vitamin D-resistant hypophosphatemic mice. J Biol Chem 266:13616–13620

    Google Scholar 

  17. Rifas L, Dawson LL, Halstead LR, Roberts M, Avioli LV (1994) Phosphate transport in osteoblasts from normal and X-linked hypophosphatemic mice. Calcif Tissue Int 54:505–510

    Google Scholar 

  18. Chen PS Jr, Toribara TY, Warner H (1956) Microdetermination of phosphorus. Anal Chem 28:1756–1758

    Google Scholar 

  19. Boyarsky G, Ganz MB, Sterzel RB, Boron WF (1988) pH regulation in single glomerular mesangial cells I. Acid extrusion in absence and presence of HCO3- Am J Physiol 255:C844–856

    Google Scholar 

  20. Thomas JA, Buchsbaum RN, Zimniak A, Racker E (1979) Intracellular pH measurements in Ehrlich ascites tumor cells utilizing spectroscopic probes generated in situ. Biochemistry 18: 2210–2218

    Google Scholar 

  21. Kurokawa K, Ohno T, Rasmussen H (1973) Ionic control of renal gluconeogenesis II. The effects of Ca2+ and H+ upon the response to parathyroid hormone and cyclic AMP. Biochim Biophys Acta 313:32–41

    Google Scholar 

  22. Kashiwagura T, Deutsch CJ, Taylor J, Erecińska M, Wilson DF (1984) Dependence of gluconeogenesis, urea synthesis, and energy metabolism of hepatocytes on intracellular pH. J Biol Chem 259:237–243

    Google Scholar 

  23. Green J, Yamaguchi DT, Kleeman CR, Muallem S (1990) Cytosolic pH regulation in osteoblasts regulation of anion exchange by intracellular pH and Ca2+ ions. J Gen Physiol 95: 121–145

    Google Scholar 

  24. Green J, Kleeman CR (1992) Role of calcium and cAMP messenger systems in intracellular pH regulation of osteoblastic cells. Am J Physiol 262:C111–1121

    Google Scholar 

  25. Graham CS, Tashjian AH Jr (1992) Mechanisms of activation of Na+/H+ exchange in human osteoblast-like SaOS-2 cells. Biochem J 288:137–143

    Google Scholar 

  26. Fliegel L, Walsh MP, Singh D, Wong C, Barr A (1992) Phosphorylation of the C-terminal domain of the Na+/H+ exchanger by Ca2+/calmodulin-dependent protein kinase II. Biochem J 282:139–145

    Google Scholar 

  27. Nesbit T, Econs MJ, Byun JK, Tenehouse HS, Drezner MK (1994) Normal Pi transport in heterogenous S1 cells of the proximal tubule (PT): evidence that an intrinsic renal abnormality does not underlie XLH. J Bone Miner Res 9(suppl 1):S146

    Google Scholar 

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Authors and Affiliations

  1. Department of Internal Medicine, Division of Bone and Mineral Diseases, Washington University School of Medicine and The Jewish Hospital of St. Louis, 216 SO. Kingshighway, 63110, St. Louis, Missouri, USA

    L. Rifas & L. V. Avioli

  2. Department of Internal Medicine, Division of Nephrology, Washington University School of Medicine and The Jewis Hospital of St. Louis, 216 SO Kingshighway 63110, 63110, St. Louis, Missouri, USA

    A. Gupta & K. A. Hruska

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  1. L. Rifas
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  2. A. Gupta
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  3. K. A. Hruska
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  4. L. V. Avioli
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Rifas, L., Gupta, A., Hruska, K.A. et al. Altered osteoblast gluconeogenesis in X-linked hypophosphatemic mice is associated with a depressed intracellular pH. Calcif Tissue Int 57, 60–63 (1995). https://doi.org/10.1007/BF00298998

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  • DOI: https://doi.org/10.1007/BF00298998

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