Glucagon-like peptide-1 (GLP-1) is released after food intake to act as an incretin. GLP-1 also inhibits gastric emptying and increases satiety. In rats, GLP-1 inhibits small bowel motility. Our aim was to study the effects of GLP-1 on gastrointestinal motility in healthy subjects and patients with irritable bowel syndrome (IBS). Antro-duodeno-jejunal manometry was carried out during a 4-h control period with saline, followed by a 4-h period with intravenous GLP-1 (healthy: 0.7 and 1.2 pmol kg−1 min−1 (n = 16); IBS, 1.2 and 2.5 pmol kg−1 min−1 (n = 14). Plasma was analysed for GLP-1 and gut hormones, and gut tissue expression of GLP-1 receptor was studied. In healthy subjects, GLP-1 0.7 pmol kg−1 min−1 reduced the migrating motor complexes (MMCs) from a median of 2 (range 2–3) to 0.5 (0–2), and motility index from 4.9 ± 0.1 to 4.3 ± 0.3 ln ∑(mmHg*s min−1) in jejunum, while GLP-1 1.2 pmol kg−1 min−1 diminshed MMCs from 2 (2–3) to 1.5 (1–2.5), and motility index from 5.2 ± 0.2 to 4.4 ± 0.2. In IBS patients, GLP-1 1.2 pmol kg−1 min−1 reduced the MMCs from 2.5 (2–3.5) to 1 (0–1.5) without affecting motility index. At 2.5 pmol kg−1 min−1 GLP-1 decreased MMCs from 2 (1.5–3) to 1 (0.5–1.5), and motility index from 5.2 ± 0.2 to 4.0 ± 0.5. Motility responses to GLP-1 were similar in antrum and duodenum. Presence of the GLP-1 receptor in the gut was verified by reverse transcriptase PCR. In conclusion, the gut peptide GLP-1 decreases motility in the antro-duodeno-jejunal region and inhibits the MMC in healthy subjects and IBS patients.