We compared the effects of four vitamin D metabolites, 1 alpha,25 dihydroxy vitamin D3 (1 alpha,25(OH)2D3), 1 alpha hydroxy vitamin D3 (1 alpha OH D3), 25 hydroxy vitamin D3 (25 OH D3), and 24R,25 dihydroxy vitamin D (24R,25(OH)sD3) on resorption and collagen synthesis in fetal rat bone maintained in organ culture. Resorption was quantitated by measuring the release of previously incorporated 45Ca from long bone shafts of 19-day fetal rats, and collagen synthesis was assessed by measuring the incorporation of 3H-proline into collagenase digestible protein (CDP) in calvaria from 21-day fetal rats. All four compounds stimulated bone resorption and inhibited collagen synthesis, but 1 alpha,25(OH)2D3 was approximately 1000 times more potent in both organ culture systems. Although the differences were small among the other three compounds, the order of potency was 1 alpha OH d3 > 25 OH D3 greater than or equal to 24R,25(OH)2D3. These results suggest that the receptor for 1 alpha 25(OH)2D3 in both bone resorbing and bone forming cells has similar affinities for several vitamin D metabolites.