Using an experimental rat model, this study was undertaken to assess the effects of a short-term application of high dose 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] on calcium homeostasis, cancellous bone formation, and numbers of osteoblast precursors in ex vivo bone marrow cultures. For Exp 1 and 2, 6-month-old female rats were sc injected with either 0.2 microg 1,25-(OH)2D3/kg x day or vehicle on days 1, 2, and 3 of the studies. Serum calcium and urinary excretion of calcium were monitored for 12 days in Exp 1. In Exp 2, the rats were ip labeled with five different fluorochromes on days 0, 5, 10, 15, and 20, respectively. Half of the rats in each group were killed on day 7, the rest of the rats were killed on day 24, and the first lumbar vertebrae were processed for histomorphometry. In Exp 3, 0.2 microg 1,25-(OH)2D3/kg BW or vehicle was sc administered to 6-month-old male rats on days 1, 2, and 3, and the number of colony-forming units with the ability to express alkaline phosphatase, to calcify, and/or to synthesize collagen were enumerated sequentially on days 4, 6, 8, 10, 12, and 14 in bone marrow cultures. Short-term 1,25-(OH)2D3 treatment resulted in increased values for serum and urinary calcium during the treatment phase in Exp 1, depressed osteoclast numbers and strongly elevated osteoblast perimeter by day 7, and stimulated mineral apposition rate and bone formation rate in the interval between days 5-15 of Exp 2. Moreover, 1,25-(OH)2D3 administration to rats significantly enhanced the number of mesenchymal precursor cells in bone marrow with the ability to differentiate into an osteoblastic phenotype in ex vivo bone marrow cultures on day 4 of Exp 3. These studies provide evidence that short-term 1,25-(OH)2D3 treatment creates new bone remodeling units and augments osteoblast recruitment and osteoblast team performance in rat cancellous bone.