Changes in muscle lipid metabolism induced in vitro by 1,25-dihydroxy-vitamin D-3

1,25-Dihydroxy-vitamin D-3 has been shown to increase phosphatidylcholine and decrease phosphatidylethanolamine levels in skeletal muscle in vitro. To elucidate the metabolic pathway and mechanism involved in the effect of the sterol, chick embryo myoblast and vitamin D-deficient chick soleus muscle cultures were treated with 1,25-dihydroxy-vitamin D-3 (5.0 X 10(-10)-3.6 X 10(-11) M, total concentration) for 12-14 h and 1 h, respectively, in the absence and presence of protein and RNA synthesis inhibitors. Lipids were then labelled with [3H]glycerol and [14C]acetate. A significant increase in phosphatidylcholine and triacylglycerol labelling and a decrease in phosphatidylethanolamine labelling were observed in response to the hormone. Cycloheximide suppressed these changes in both types of preparations. Puromycin and actinomycin D were also effective blockers in cultured muscle cells. In addition, double-labelling of control and 1,25-dihydroxy-vitamin D-3-treated myoblasts with [3H]choline and [14C]ethanolamine were performed. The sterol did not affect [3H]choline labelling of total cell lipid extracts and phosphatidylcholine. However, the total lipid fraction of treated cells was labelled to a greater extent with [14C]ethanolamine. In addition, an increased incorporation of this precursor into phosphatidylcholine accompanied by a proportional decrease in phosphatidylethanolamine could be shown in cells pretreated with the metabolite. These changes were abolished by cycloheximide and actinomycin D. The results suggest that 1,25-dihydroxy-vitamin D-3 stimulates methylation of phosphatidylethanolamine into phosphatidylcholine in myoblasts by a nuclear mechanism. The data are consistent with the presence in the cells of a receptor specific for the hormone.