Effects of starvation and different culture conditions on the phospholipid content of isolated pancreatic islets

Abstract

A colorimetric method for the determination of lipid phosphorus in the nanomolar range was used to determine the total phospholipid content of isolated pancreatic islets. Freshly isolated islets of lean C57BL/6J mice contained significantly more phospholipids expressed per μg DNA as compared to C57BL/6J (ob/ob) mouse or Wistar rat islets. Starvation for 48 h (Wistar rats) or 60 h (NMRI mice) did not affect the islet phospholipid content. Phosphatidylcholine was the most abundant phospholipid class of NMRI mouse islets, followed by phosphatidylethanolamine, sphingomyelin, phosphatidylinositol, phosphatidylserine and lysophosphatidylcholine. When islets of NMRI mice were maintained for 5–7 days in tissue culture, the phospholipid content remained unchanged as compared to that of freshly isolated islets despite a considerable loss of the insulin stores. The islet phospholipid content was significantly increased when the glucose concentration of the culture medium was elevated from 3 to 28 mM. Leucine (10 mM) added to a low-glucose medium failed to increase the islet phospholipid content. Addition of glipizide (2 μM) to the culture medium decreased the islet insulin content significantly but failed to affect the total islet phospholipid content. Culture in a Ca 2+-free medium containing 28 mM glucose increased the islet insulin content but, again, the phospholipid content remained unaffected. These data show that changes of the total phospholipid content of pancreatic islets are unrelated to the islet insulin content and presumably also to the content of secretory granules. Alterations of the islet content of phospholipids may rather reflect changes of the amount of endoplasmic reticulum of the islet cells.