Glucose transport, phosphorylation, and utilization in isolated porcine pancreatic islets.
Porcine islets have been proposed as a donor source for human transplantation, mainly because of both structural and biological similarities of porcine and human insulin. However, the in vitro function of these islets is poorly characterized. In the present study, we first examined insulin release in response to glucose in static incubation experiments. Increasing glucose concentrations up to 8.3 mmol/L stimulated insulin release; however, this elevation was only twofold, and a paradoxical decline was observed at glucose concentrations higher than 8.3 mmol/L. In cultured porcine islets, a greater insulin secretion may be elicited by agents that increase intracellular cyclic adenosine monophosphate (cAMP) levels. To investigate the possible reasons for the porcine islet low response to glucose in vitro, we then evaluated in parallel experiments glucose transport, phosphorylation, and utilization. Glucose transport studies (using 3-O-methyl glucose uptake at 15 degrees C for 15 seconds) indicated the presence of both a high-affinity (Km, 1.2 +/- 0.6 mmol/L) and a low-affinity (Km, 11.8 +/- 1.9 nmol/L, n = 5) component. Glucose phosphorylation, evaluated by measuring the rate of glucose-6-phosphate formation in a fluorimetric assay, indicated that glucokinase activity had a maximum (Vmax) of 7.97 +/- 0.94 nmol/microgram DNA/h and a Km of 8.3 +/- 0.9 mmol/L (mean +/- SE, n = 8).(ABSTRACT TRUNCATED AT 250 WORDS)
Rabuazzo
A M
AM
Institute of Internal Medicine, Metabolism, and Endocrinology, University of Catania Medical School, Italy.
Davalli
A M
AM
Buscema
M
M
Socci
C
C
Caltabiano
V
V
Pontiroli
A E
AE
Di Carlo
V
V
Pozza
G
G
Vigneri
R
R
Purrello
F
F
D000818:Animals; D001692:Biological Transport; D003470:Culture Media; D000242:Cyclic AMP / Q000378:metabolism; D005947:Glucose / Q000378:metabolism* / Q000494:pharmacology; D066298:In Vitro Techniques; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000378:metabolism*; D009004:Monosaccharide Transport Proteins / Q000378:metabolism; D010766:Phosphorylation; D013552:Swine
D003470:Culture Media; D007328:Insulin; D009004:Monosaccharide Transport Proteins; D000242:Cyclic AMP; D005947:Glucose