Summary
To determine the relationship between alkaline phosphatase (AP), 1,25(OD)2D3 and bone formationin vivo, we have examined the effects of levamisole, a stereospecific inhibitor of AP on bone formation and on 1,25(OH)2D3-stimulated bone mineralization in the mouse. Normal mice were injected daily with levamisole at doses of 40 and 80 mg/kg/b.w. The compound was given alone or in combination with 1,25(OH)2D3 infusion (0.05 μg/kg/d) for 7 days. Treatment with levamisole alone inhibited the serum AP activity (mainly of skeletal origin in mice) by 18.4 and 61.3% for the low and high dose respectively. No deleterious effect on body growth, tibia length, and bone cells population was detected. The moderate inhibition of AP activity produced by the lower dose of levamisole alone (18.4%) or in combination with 1,25(OH)2D3 (37.9%) was associated with a reduced endosteal matrix apposition rate (MaAR) determined by double3H-proline labeling method. This effect was related to a levamisole-induced fall in serum phosphate. Despite the moderate inhibition of AP activity, the mineral apposition rate (MiAR) determined by the double tetracycline labeling method remained normal. Moreover, 1,25(OH)2D3 infusion still resulted in increased MiAR which was stimulated to the same extent as in the absence of levamisole. By contrast, the more severe inhibition of AP activity induced by 80 mg/kg of levamisole alone (61.3%) or in combination with 1,25(OH)2D3 (45.8%) inhibited both the MaAR and the MiAR and prevented the stimulatory effect of 1,25(OH)2D3 on bone mineralization. The data show that AP activity affects the bone matrix and mineral apposition ratesin vivo and that severe inhibition of AP activity inhibits the 1,25(OH)2D3-induced stimulation of bone mineralization in the mouse.
Similar content being viewed by others
References
Whyte MP (1983) Alkaline phosphatase and the measurement of bone formation. In: Frame B, Potts JT (eds) Clinical disorders of bone and mineral metabolism. Excerpta Medica, Amsterdam, Oxford, Princeton, pp 120–124
Robison R (1923) The possible significance of hexosephosphoric esters in ossification. Part I. Biochem J 17:268–293
Ali SY, Sajdera SW, Anderson HC (1970) Isolation and characterization of calcifying matrix vesicles from epiphyseal cartilage. Proc Natl Acad Sci 67:1513–1520
Van Belle H (1976) Alkaline phosphatase. I-kinetics and inhibition by levamisole of purified isoenzymes from humans. Clin Chem 22:972–976
Thomas MC, Ramp WK (1979) Effects of parathyroid hormone on alkaline phosphatase activity and mineralization of cultured chick embryo tibia. Calcif Tissue Int 27:137–142
Nijweide PJ, Kawilarang De Haas EWM, Wassenaar AM (1981) Alkaline phosphatase and calcification, correlated or not? Metab Bone Dis Rel Res 3:61–66
Fallon M, Whyte M, Teitelbaum S (1980) Stereospecific inhibition of alkaline phosphatase by levamisole prevents in vitro calcification of rachitic rat cartilage. Lab Invest 43:489–494
Farley JR, Puzas JE, Baylink DJ (1982) Effect of skeletal alkaline phosphatase inhibitors on bone cell proliferation in vitro. Mineral Electrolyte Metab 7:316–323
Manolagas SC, Burton DW, Deftos LJ (1981) 1,25-dihydroxyvitamin D3 stimulates the alkaline phosphatase activity of osteoblast-like cells. Biol Chem 256:7115–7117
Mulkino MA, Manolagas SC, Deftos LJ, Sussman HH (1983) 1,25-Dihydroxyvitamin D3 increases bone alkaline phosphatase isoenzyme levels in human osteogenic sarcoma cells. J Biol Chem 258:6219–6225
Kumegawa M, Ikeda E, Tanaka S, Haneji T, Yora T, Sakagishi Y, Minami N, Hiramatsu M (1984) The effects of prostaglandin E2, parathyroid hormone, 1,25-dihydroxycholecalciferol and cyclic nucleotide analogs on alkaline phosphatase activity in osteoblastic cells. Calcif Tissue Int 36:72–76
Majeska RJ, Rodan GA (1982) The effect of 1,25(OH)2D3 on alkaline phosphatase in osteoblastic osteosarcoma cells. J Biol Chem 257:3362–3365
Borgers M (1973) The cytochemical application of new potent inhibitors of alkaline phosphatase. J Histochem Cytochem 21:812–824
Marie PJ, Travers R (1983) Continuous infusion of 1,25-dihydroxyvitamin D3 stimulates bone turnover in the normal young mouse. Calcif Tissue Int 35:418–425
Marie PJ, Hott M, Garba MT (1985) Contrasting effects of 1,25 dihydroxyvitamin D3 on bone and mineral appositional rates in the mouse. Metabolism 34:777–783
Shepard RM, Deluca HF (1980) In: Cormic DB, Wright LD (eds) Methods in enzymology: determination of vitamin D and its metabolites in plasma. Academic Press, New York pp 393–414
Menahan LA, Sobocinski KA, Austin BP (1984) The origin of plasma alkaline phosphatase activity in mice and rats. Comp Biochem Physiol 79B:279–283
Marie PJ (1982) Dissertation thesis, University of Paris VI, Paris
Borgers M, Thone F (1975) The inhibition of alkaline phosphatase by L.p- bromotetramisole. Histochemistry 44:277–280
Rasmussen H (1983) Hypophosphatasia. In: Stanbury JB, Wingaarden JB, Frederickson DS, Goldstein JL, Brown MS (eds) The metabolic basis of inherited disease, 5th ed. McGraw-Hill, New York, pp 1497–1507
Lerner V, Granstrom G (1984) Effect of levamisole on bone resorption in cultured mouse calvaria. Eur J Pharmacol 105:1–14
Wong GL, Luben RA, Cohn DV (1977) 1,25 Dihydroxycholecalciferol and parathormone: effects on isolated osteoclast-like and osteoblast-like cells. Science 197:663–665
Canalis EM (1983) Effect of hormones and growth factors on alkaline phosphatase activity and collagen synthesis in cultured rat calvariae. Metabolism 32:14–20
Haneji T, Kurihara N, Ikeda K, Kumegawa M (1983) 1α,25-Dihydroxyvitamin D3 and analogues of vitamin D3 induce alkaline phosphatase activity in osteoblastic cells derived from newborn mouse calvaria. J Biochem 94:1127–1132
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Garba, MT., Marie, P.J. Alkaline phosphatase inhibition by levamisole prevents 1,25-dihydroxyvitamin D3-stimulated bone mineralization in the mouse. Calcif Tissue Int 38, 296–302 (1986). https://doi.org/10.1007/BF02556610
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF02556610