Titre du document / Document title

Glucose transport, phosphorylation, and utilization in isolated porcine pancreatic islets

Auteur(s) / Author(s)

RABUAZZO A. M. ; DAVALLI A. M. ; BUSCEMA M. ; SOCCI C. ; CALTABIANO V. ; PONTIROLI A. E. ; DI CARLO V. ; POZZA G. ; VIGNERI R. ; PURRELLO F. ;

Affiliation(s) du ou des auteurs / Author(s) Affiliation(s)

Univ. Catania medical school, inst. internal medicine metabolism endocrinology, Catania, ITALIE

Résumé / Abstract

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^oC for 15 seconds) indicated the presence of both a high-affinity (K_m, 1.2±0.6 mmol/L) and a low-affinity (K_m, 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/μg DNA/h and a K_m of 8.3±0.9 mmol/L (mean±SE, n=8). Glucose utilization, determined by measuring ³H₂O formation from (5-³H)-glucose, increased from 1.79±0.34 nmol/μg DNA/2 h at 5 mmol/L glucose to 5.97±0.16 and 6.19±0.29 at 20 and 30 mmol/L glucose, respectively, with a calculated Vmax of 6.9±0.33 and a K_m of 7.0±1.4 mmol/L (mean±SE, n=5). These data in porcine islets were similar to data previously obtained in rat islets using the same procedures. In conclusion, this study indicates that glucose transport, phosphorylation, and utilization are not altered in isolated porcine islets, and therefore, the in vitro low response of porcine islets to glucose is not due to alterations in glucose sensing or metabolism

Revue / Journal Title

Metabolism, clinical and experimental   ISSN 0026-0495 

Source / Source

1995, vol. 44, n^o2, pp. 261-266 (20 ref.)

Langue / Language

Anglais

Editeur / Publisher

Elsevier, New York, NY, ETATS-UNIS  (1952) (Revue)

Mots-clés anglais / English Keywords

Glucose ; Biological transport ; Phosphorylation ; Langerhans islet ; Pancreas ; RP-Insulin ; Animal model ; In vitro ; Animal ; Pig ; Artiodactyla ; Ungulata ; Mammalia ; Vertebrata ; Endocrine pancreas ; Hormone releasing factor ; Gastrointestinal hormone ;

Mots-clés français / French Keywords

Glucose ; Transport biologique ; Phosphorylation ; Ilot Langerhans ; Pancréas ; Insuline RP ; Modèle animal ; In vitro ; Animal ; Porc ; Artiodactyla ; Ungulata ; Mammalia ; Vertebrata ; Pancréas endocrine ; Facteur libération hormonale ; Hormone gastrointestinale ;

Mots-clés espagnols / Spanish Keywords

Glucosa ; Transporte biológico ; Fosforilación ; Isla Langerhans ; Páncreas ; Insulina RP ; Modelo animal ; In vitro ; Animal ; Cerdo ; Artiodactyla ; Ungulata ; Mammalia ; Vertebrata ; Páncreas endocrino ; Factor liberación hormonal ; Hormona gastrointestinal ;

Localisation / Location

INIST-CNRS, Cote INIST : 6965, 35400005624219.0220