The secretion and molecular nature of immunoreactive neurotensin (NT) was studied following stimulation of an isolated perfused porcine ileal segment with glucose, triglyceride and intra-arterial infusion of gastrin-releasing peptide (GRP). Secreted peptides were separated using gel chromatography and analyzed with 3 sequence-specific radioimmunoassays towards NT. Glucose (5%) and GRP both stimulated NT secretion from the ileal segment whereas pure triglyceride did not. Maximal secretion of NT during glucose perfusion was 0.448 nmol/min and 6.9 nmol/min during GRP infusion (medians, n = 5). GRP infused in doses from 10(-10) to 10(-8) M stimulated NT release in a dose-related manner. Following gel chromatography only the intact peptide and no smaller or larger molecular size immunoreactive components were observed. The study showed that both luminal and humoral stimuli release NT from the isolated pig ileum. Apparently no fragments or other NT-related immunoreactive components were cosecreted with the peptide.