Vitamin D and enterocyte brush border membrane calcium transport and fluidity in the rat

Prior studies of vitamin D repletion showed a threefold increase in the maximum rate (Vmax) for calcium uptake by brush border membrane vesicles, but did not differentiate saturable and nonsaturable uptake components. We studied the calcium uptake and fluidity response of intestinal brush border vesicles to vitamin D by treatment with 1 alpha,25-dihydroxy-24,24-difluorocholecalciferol (24,24-F-1,25-(OH)2D3). Treatment responses were measured by effects on (1) saturable and nonsaturable initial uptake rates of calcium by rat proximal small intestinal brush border membrane vesicles; (2) transmucosal calcium transport by everted duodenal sac; and (3) fluorescence anisotropy. Treatment of vitamin D-depleted weanlings increased the Vmax by 50% (P < .05) in vesicles from the proximal 12 cm of small intestine from rats injected with disodium ethane-1-hydroxy-1,1-diphosphonate (EHDP), but there was no response in rats not injected with EHDP or in vesicles from the proximal 30 cm of small intestine. Vitamin D-depleted weanlings were D-deficient based on serum 25-hydroxycalciferol(25-OH-D) concentration, but to produce 1 alpha,25-dihydroxycalciferol [1,25-(OH)2D] depletion, EHDP injection was required. Treatment of vitamin D-replete adult rats caused a 20% (P < .05) increase in Vmax. Treatment did not affect the calcium concentration at half-Vmax (KT), the rate constant for nonsaturable uptake (KD), or vesicle fluidity measured as fluorescence anisotropy. Contrasting with these minimal effects of treatment on brush border Vmax, treatment increased transmucosal calcium transport by everted duodenal sac almost threefold in vitamin D-depleted weanlings administered EHDP. Thus, vitamin D actions on enterocyte calcium transport (1) at the brush border increase saturable but not nonsaturable uptake, and (2) produce the major transport response distal to the brush border. Despite previously described changes in membrane lipid, brush border fluidity is unaffected by vitamin D treatment.