Pancreatic lipid and fatty acid composition was determined in streptozotocin-induced diabetic rats and compared to control and insulin-treated diabetic rats. A pronounced decrease of total fatty acids was recorded and mainly accounted for by a drop of fatty acids in glycerides. Cholesterol, on the other hand, was significantly increased two- to three-fold, leading to an elevated cholesterol/phospholipid ratio. Morpho-cytochemical studies confirmed these findings because the multiple lipid droplets present in acinar cells of diabetic animals were found to be of cholesterol nature. The major alterations in phospholipid-fatty acid composition were characterized by an increase of linoleate coupled to a decrease of monounsaturates and arachidonate, suggesting defective metabolism of saturates and of linoleate. This was further supported by fatty acid ratios that suggested low delta 5 and delta 9 desaturation. Daily administration of insulin for 10 days restored and overcorrected the various lipid alterations. This study suggests that there are alterations in lipid composition and metabolism in the exocrine pancreas of chronic streptozotocin-induced diabetic rats. Their possible role in the mechanism regulating pancreatic function and secretion remains to be elucidated.