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In vitro effects of vitamin D3 metabolites on rat calvaria cAMP content

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Summary

Results from in vitro works suggest that 1,25- and 24,25-dihydroxyvitamin D3 (1,25-(OH)2D3 and 24,25-(OH)2D3) act on bone via different mechanisms. The present investigation was performed to study the effect of these two metabolites and of their precursor 25-hyxdroxyvitamin D3 (25-(OH)D3) on bone cAMP content in vitro. Rats' paired half calvaria were incubated under sterile conditions with one vitamin D3 derivative (10−13 to 10−9 M) or with ethanol (0.005 ml for 15 min to 24 h in 1 ml medium containing 0, 0.2, 1, 2, or 3 mM calcium. In some experiments: (a) cycloheximide (10−5M) was added simultaneously with the vitamin D3 metabolites; (b) 1–84 bPTH (5 × 10−8 M) was added for 5 or 15 min at the end of the 24 h incubation. Calvaria were immersed in 1 ml TCA 5% 4°C and homogenized. The cAMP was extracted with diethylether and measured by a competitive protein binding assay. Results bring further evidence for a particular effect of low doses of 24,25-(OH)2D3 (10−9 to 10−12M) and of 25-(OH)D3 (10−9 to 10−11M) on bone, different from that of 1,25-(OH)2D3: cAMP content was higher in 24,25-(OH)2D3- or 25-(OH)D3-treated and lower in 1,25-(OH)2D3-treated calvaria than in ethanol-treated ones with 1 mM calcium. The 1,25-(OH)2D3 effect persisted in calcium-free medium whereas 25-(OH)D3 and 24,25-(OH)2D3 effects could not be observed with 0 mM nor with 3 mM calcium. The required duration of the preincubation (over 1 h) as well as the inhibitory action of cycloheximide may suggest an involvement of protein synthesis in the vitamin D3 metabolites effects. Neither 1,25-(OH)2D3 nor 24,25-(OH)2D3 affected the PTH-induced increase in bone cAMP content.

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Authors and Affiliations

  1. Laboratoire des Tissus Calcifiés (CNRS ER 126), Unité de Recherche sur les Maladies du Métabolisme chez l'Enfant (INSERM U.30), Hôpital des Enfants-Malades, Tour Technique 6ème étage, 149 rue de Sèvres, 75730, Paris Cédex 15, France

    M. Lieberherr, M. Garabédian, H. Guillozo, C. L. Thil & S. Balsan

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  1. M. Lieberherr
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  2. M. Garabédian
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  3. H. Guillozo
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  4. C. L. Thil
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  5. S. Balsan
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Lieberherr, M., Garabédian, M., Guillozo, H. et al. In vitro effects of vitamin D3 metabolites on rat calvaria cAMP content. Calcif Tissue Int 30, 209–216 (1980). https://doi.org/10.1007/BF02408630

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  • DOI: https://doi.org/10.1007/BF02408630

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