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A possible mechanism of cadmium-copper interaction in embryonic chick bone in tissue culture
Abstract:
Femurs from 9-day-old embryo were cultured for 4 days by the roller-tube method in the presence of Cd and/or Cu. The combination of both Cd and Cu caused a significantly interactive decrease in hydroxyproline (Hyp) synthesis as well as bone growth compared with that in the presence of Cd (1.1 or 3.3 microM) or Cu (1.1 or 2.2 microM) alone. The presence of both 2.2 microM Cd and 3.3 microM Cu also showed a significantly interactive decrease in the incorporation of [3H]proline (Pro) into collagenase-digestible protein (CDP), but it showed no interactive inhibition of the hydroxylation of [3H]Pro in CDP. The two metals were also interactive with respect to the inhibition of the synthesis of protein, RNA and DNA. Culture of epiphysis in the presence of both Cd and Cu resulted in higher content of Cd and Cu compared to those cultured in Cd or Cu alone. Subcellular fractions from epiphysis cultured in Cd plus Cu contained more Cd than those cultured in Cd alone. Cu was increased in two fractions, nuclei and cytosol, following co-incubation. The cytosol from epiphysis cultured in the presence of both Cd and Cu contained more Cd in both a metallothionein (MT)-like protein and a high-molecular-weight (HM) protein than cytosol from Cd-treated bones. The amount (nmol) of Cu in the MT fraction was nearly equal to the sum of the increased amounts (nmol) of Cd in HM fraction of cytosol and particulate fractions. This indicates that some Cd in MT-like protein induced by Cd is replaced with Cu and the released Cd redistribution to HM fraction and particulate fractions. Most of the Cd in the solubilized particulate fractions was detected in the HM fraction. A marker enzyme or component in each fraction was interactively inhibited by both Cd and Cu. Therefore, the interactive inhibition of bone metabolism by both Cd and Cu in the cultured bone is at least partly due to the increase in Cd content of HM fraction of cytosol and particulate fractions.