Either vitamin D3 (or 1α,25-(OH)2-D3) or hydrocortisone (HC) stimulated phosphate accumulation by organ-cultured embryonic chick duodenum. In combination, these two steroids stimulated phosphate uptake synergistically. Phosphate accumulation appeared to be independent of other vitamin D3-stimulated processes: CaBP concentration, cAMP concentration, or alkaline phosphatase activity. L-phenylalanine, a reported alkaline phosphatase inhibitor, when added to the culture medium progressively inhibited either D3- or HC-stimulated phosphate uptake subsequent to culture, but did not inhibit the synergistic action. Under these conditions L-phenylalanine had no consistent effect on alkaline phosphatase activity but unexpectedly, greatly inhibited vitamin D3-stimulated CaBP concentration, but only in the absence of HC. Some limited suggestion of an intestinal phosphoprotein sensitive to either vitamin D3 or HC was observed.