OBJECTIVE Rerouting of nutrients and/or increasing nutrient delivery to the small intestine after Roux-en-Y gastric bypass may have important potential as a diabetes treatment modality. However, it is still important question which part of the gastrointestinal tract is the most important for control of glycemia. The aim of this study was to investigate the role of different segments of the gastrointestinal tract on glucose metabolism in the physiological state. METHODS Forty 12-week-old male Wistar rats were divided into the following four groups of 10 animals each: the gastrostomy group, the duodenostomy group, the jejunostomy group, and the ileostomy group. All rats were subjected to a glucose tolerance test by infusion of glucose via the surgically inserted tubes in the stomach (gastrostomy), in the duodenum (duodenostomy), in the jejunum (jejunostomy), or in the ileum (ileostomy). Plasma glucagon-like peptide-17-36 (GLP-17-36) and insulin levels during the glucose tolerance test were assayed and Matsuda index was calculated. RESULTS Ileostomy rats exhibited significantly lower glycemic excursions compared with gastrostomy, duodenostomy, and jejunostomy rats. Insulin and GLP-1 levels during the glucose tolerance test were significantly higher in duodenostomy and jejunostomy rats than in gastrostomy and ileostomy rats. Matsuda index was significantly higher in ileostomy rats than in duodenostomy and jejunostomy rats. CONCLUSIONS Ileal glucose infusion leads to increased insulin sensitivity, further decreasing blood glucose levels.