Background: Absorption of non- haeme iron occurs mainly in the duodenum. It involves the divalent metal transporter 1 ( DMT1) in the uptake of ferrous Fe( II) iron and the basolateral transporter ferroportin/ IREG- 1/ MTP- 1/ SLC40A1 in its release. Whether ferroportin functions in this process at other sites in the enterocyte is unknown. In this study the effect of a blocking antibody to ferroportin on the uptake and release of iron was evaluated in enterocyte- like cells ( IEC- 6 and Caco- 2) and in freshly isolated duodenal enterocytes from rats.Methods: Uptake of 1 muM Fe( II) and its release by cells was studied in the presence of the antibody. Ferroportin expression was determined by western blot analysis of duodenal mucosa enriched microvillus membranes, Caco- 2 cells, IEC- 6 cells, and freshly isolated enterocytes. Immunofluorescent detection of ferroporitn was performed on frozen sections of duodenum from rats with variations in body iron stores.Results: Ferroportin was expressed in all cell types. In these cells, the antibody significantly reduced ( p < 0.05) uptake of Fe( II) by 40 - 50% but had no effect on the release of iron. In Caco- 2 cells, Fe( II) uptake was reduced only when the antibody was in contact with the apical membrane. Ferroportin protein was enriched in microvillus membrane preparations. In enterocytes from iron deficient rats, ferroportin was expressed along the brush border where it colocalised with lactase. Ferroportin was seen in the basal cytoplasm and along the basolateral membranes. Iron loading markedly reduced intracellular expression of ferroportin. In Caco- 2 cells, ferroportin also localised to the microvillus and lateral and basal membranes.Conclusions: In addition to release, ferroportin functions in the uptake of iron at the apical membrane, possibly by modulating the activity of DMT1.