The physiological significance of the presence of GLUT2 at the food-facing pole of intestinal cells, is addressed by a study of fructose absorption in GLUT2-null and control mice submitted to different sugar diets. Confocal microscopy localization, protein and mRNA abundance as well as tissue and membrane vesicle uptakes of fructose were assayed. GLUT2 was located in the basolateral membrane of mice fed a meal devoid of sugar or containing complex carbohydrates. In addition, the ingestion of a simple sugar meal promoted the massive recruitment of GLUT2 to the food-facing membrane. Fructose uptake in brush border membrane vesicles from GLUT2-null mice was half that of wild-type mice and was similar to the cytochalasin B-insensitive component i.e. GLUT5-mediated uptake. A 5-day consumption of sugar-rich diets, increased fructose uptake 5-fold in wild type tissue rings when it only doubled in GLUT2-null tissue. GLUT5 was estimated to contribute to 100% of total uptake in wild-type mice fed low-sugar diets, dropping down to 60% and 40% with glucose- or fructose-diets respectively; the complement was ensured by GLUT2 activity. The results indicate that basal sugar uptake is mediated by the resident food facing SGLT1, GLUT5 transporters which vary only 2-fold by long-term dietary adaptation. We also observe that a large improvement of intestinal absorption is promoted by the transient recruitment of food facing GLUT2, induced by the ingestion of a simple-sugar meal. Thus, GLUT2 and GLUT5 could exert complementary roles in adapting the absorption capacity of the intestine to occasional, or repeated loads of dietary sugars.