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The influence of plasma from genetically obese (ob/ob) and lean (+/+) mice on insulin secretion has been studied by perifusion of collagenase-prepared pancreatic islets maintained for 48 h in culture. Insulin secretion was measured at 2-min intervals and plasma from the ob/ob mice not from the +/+ mice rapidly stimulated insulin release, reaching a maximum in 2–4 min and falling to basal levels in about 10 min. Experimental evidence is given indicating that the plasma insulin secretagogue is identical to β-cell-trophin, a peptide of the pituitary pars intermedia which stimulates insulin secretion. The evidence is based on (1) the antigenic properties of the peptides (both cross-react with a -COOH terminal ACTH antiserum raised to the 17–39 moiety of ACTH), (2) identical chromatographic separation on Biogel columns and on reverse-phase high pressure liquid chromatography and (3) the similarity of their insulin releasing action from perifused islets.