Endothelial cell signalling supports pancreatic beta cell function in the rat.

AIMS/HYPOTHESIS: The proximity of endothelial cells and beta cells in islets by necessity means that they are exposed to each other's products. Whereas islet endothelial cells require signals from beta cells to function properly, endothelin-1, thrombospondin-1 and laminins, among others, have been identified as endothelial-derived molecules, although their full effects on beta cells have not been explored. We tested the hypothesis that islet endothelial-derived products affect beta cell function. METHODS: Endothelial cells from rat islets were proliferated and purified. Endothelium-conditioned culture medium (ECCM) was obtained by maintaining the endothelial cells in culture medium. Islet function was evaluated following exposure of cultured islets to standard culture medium or ECCM. Changes in mRNA levels for key beta cell metabolic enzymes were also measured in islets after ECCM exposure. RESULTS: Glucose-stimulated insulin release and islet insulin content were markedly enhanced by exposure to ECCM. This was at least partly explained by improved mitochondrial function, as assessed by glucose oxidation and an upregulation of the mitochondrial gene for glycerol-3-phosphate dehydrogenase (mGpdh [also known as Gpd2]), combined with upregulation of the rate-limiting enzyme in the glycolysis, glucokinase, in the islets. The intracellular degradation of insulin was also decreased in the islets. Islet endothelial cells produced laminins, and the positive effects of islet endothelial cells were prevented by addition of a neutralising antibody to the beta1-chain of laminin. Addition of exogenous laminin stimulated islet function. CONCLUSIONS/INTERPRETATION: This study provides proof of principle that endothelial cells can affect the function of beta cells in their vicinity and that this is at least partially mediated by laminins.

Alterations in net glucose uptake and in the pancreatic B-cell GLUT2 transporter induced by diazoxide and by secretory stimuli.

The pancreatic B-cell GLUT2 transporter and glucose metabolism were examined in isolated rat islets subjected to treatments affecting insulin secretion. Diazoxide was used to inhibit, while glipizide or depolarization of the plasma membrane with a high extracellular K(+) concentration were used to stimulate insulin release in short-term experiments. Islet GLUT2 and insulin were determined by quantitative immunohistochemistry and GLUT2 was also determined by Western blot analysis. Islet net glucose uptake and glucose oxidation were measured using radioactively labelled glucose. Exposure of the islets to diazoxide was associated with a marked increase in the B-cell plasma membrane staining for GLUT2 and increased net glucose uptake. Glucose oxidation was not changed, which may reflect a lowered energy requirement. Conversely, islets subjected to a stimulated insulin secretion with glipizide or a high extracellular K(+) concentration showed a reduced staining of the GLUT2 transporter. The net glucose uptake and glucose oxidation were also reduced. In islets exposed to the high K(+) concentration no change in the molecular weight or phosphorylation of GLUT2 was observed but a lesser amount of the transporter was found by Western blot analysis. Thus, GLUT2 and glucose uptake in the pancreatic B-cell are modified by the secretory process, which suggests that changes in the glucose transporter have a functional role in normal B-cell physiology.