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. 1971 Nov;125(1):147–153. doi: 10.1042/bj1250147

The localization of 1,25-dihydroxycholecalciferol in bone cell nuclei of rachitic chicks

J C Weber 1,*, V Pons 1,*, E Kodicek 1
PMCID: PMC1178034  PMID: 4333933

Abstract

1. A simple technique has been developed to obtain subcellular fractions of chick bone. The method yielded 60–70% of total DNA in the nuclear debris fraction and 80–90% of total 14C recovered in bone after a dose of radioactive vitamin D. 2. After a dose of [4-14C,1,2-3H2]cholecalciferol (0.5μg) was given to vitamin D-deficient chicks, the time-course of total 14C radioactivity in the epiphysis, metaphysis and diaphysis of proximal tibiae was measured. The maximum concentrations were reached at 6h, corresponding to a similar peak of radioactivity in blood, decreasing until 24h and indicating the dependence on the circulating 14C and on the blood supply of the three bone components. 3. The 14C radioactivity of cholecalciferol and 25-hydroxycholecalciferol (expressed per mg of DNA) followed the pattern of incorporation of total 14C radioactivity in all three bone components. The more polar metabolite fraction reached a peak of radioactivity at 6–9h and maintained its concentration over the 24h period studied in all anatomical bone components. 4. After a dose of [4-14C,1-3H]cholecalciferol (0.5μg) was given to vitamin D-deficient chicks, the subcellular distribution was studied. At 24h after dosing, the nuclear fraction contained 27% and the supernatant fraction had 67% of total 14C recovered in the bone filtrate. When the 14C in the residual bone fragments was included, the nuclear fraction contained up to 35% of the total radioactivity in the bone. 5. The subcellular distribution pattern of individual vitamin D metabolites indicated that the purified nuclear fraction concentrated the polar metabolite, which lost 3H at C-1, so that 77% of the radioactivity could be accounted for by 1,25-dihydroxycholecalciferol. The supernatant fraction contained smaller amounts of 1,25-dihydroxycholecalciferol (9%), with 66% of 25-hydroxycholecalciferol forming the major metabolite, corresponding to its concentration found in blood at 24h. 6. The preferential accumulation of 1,25-dihydroxycholecalciferol in the nuclear fraction and the overall pattern of other metabolites, found previously in intestinal tissue, suggests a similar mechanism of action in bone to that postulated for the intestinal cell in calcium translocation.

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Selected References

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