Previous reports have shown that in avian and rodent isolated skeletal muscle cells and cultured myoblast cell lines, vitamin D 3 metabolites, such as 25-hydroxyvitamin D 3 (25(OH)D 3 ) and 1␣,25(OH) 2 D 3 , influence cellular calcium and phosphorus uptake, cellular growth, differentiation, and the expression of a limited number of genes (14 -19). Many reports suggest that the vitamin D receptor (VDR) is expressed in skeletal muscle (20 -24), and VDR deletion in mice results in alterations in muscle function and strength (25,26). Treatment of vitamin D-deficient humans with cholecalciferol improves muscle phosphocreatine recovery after exercise (27), suggesting that vitamin D 3 or its metabolites alter skeletal muscle oxidative capacity.To assess the mechanism of action of the active metabolite of vitamin D 3 , 1␣,25(OH) 2 D 3 , in human skeletal muscle cells, we examined changes in mitochondrial oxygen consumption (OCR), mitochondrial dynamics, mitochondrial OXPHOS proteins, pyruvate dehydrogenase phosphorylation, and nuclear gene expression using whole transcriptome shotgun sequencing (WTSS, RNA-seq) of messenger RNAs and micro-RNAs Tables 1 and 2