1,25-Dihydroxyvitamin C3-stimulated active uptake of phosphate by rat jejunum

An assay system for measuring an active phosphate uptake in rat jejunal disks has been developed and used to study the rat's response to 1,25-dihydroxyvitamin D3. The active component of phosphate uptake is stimulated by added KCl, fructose, sodium, and CaCl2, but not MgCl2. Half-saturation values obtained with this assay are 0.08 mM (1,25-dihydroxyvitamin D3-treated) and 0.11 mM (D-deficient). The maximal velocities are 9.6 nmol . cm-2 . min-1 (D-deficient). Using a 5-min preincubation and a 15-min incubation, a dose-response curve for 1,25-dihydroxyvitamin D3 shows 6 pmol to be the lowest dose to give a significant response and 60 pmol to be the level at which the maximal response is reached. Measurement of phosphate uptake versus time after dosing with 1,25-dihydroxyvitamin D3 (300 pmol) revealed a biphasic response with peaks at 8 and 36 h and a trough at 20 h.