Glucose-regulated anaplerosis and cataplerosis in pancreatic beta-cells: possible implication of a pyruvate/citrate shuttle in insulin secretion.
The hypothesis proposing that anaplerosis and cataplerosis play an important role in fuel signaling by providing mitochondrially derived coupling factors for stimulation of insulin secretion was tested. A rise in citrate coincided with the initiation of insulin secretion in response to glucose in INS-1 beta-cells. The dose dependence of glucose-stimulated insulin release correlated closely with those of the cellular contents of citrate, malate, and citrate-derived malonyl-CoA. The glucose-induced elevations in citrate, alpha-ketoglutarate, malonyl-CoA, and the 3-[4,5-dimethylthiazol-2yl]-2,5-diphenyltetrazolium reduction state, an index of beta-cell metabolic activity, were unaffected by the Ca2+ chelator EGTA. Glucose induced a rise in both mitochondrial and cytosolic citrate and promoted efflux of citrate from the cells. The latter amounted to approximately 20% of glucose carbons entering the glycolytic pathway. Phenylacetic acid, a pyruvate carboxylase inhibitor, reduced the glucose-induced rise in citrate in INS-1 cells and insulin secretion in both INS-1 cells and rat islets. The results indicate the feasibility of a pyruvate/citrate shuttle in INS-1 beta-cells, allowing the regeneration of NAD+ in the cytosol and the formation of cytosolic acetyl-CoA, malonyl-CoA, and NADPH. The data suggest that anaplerosis and cataplerosis are early signaling events in beta-cell activation that do not require a rise in Ca2+. It is proposed that citrate is a signal of fuel abundance that contributes to beta-cell activation in both the mitochondrial and cytosolic compartments and that a major fate of anaplerotic glucose carbons is external citrate.
Farfari
S
S
Department of Nutrition, University of Montreal, the Centre de Recherches du CHUM and Institut du Cancer, Quebec, Canada.
Schulz
V
V
Corkey
B
B
Prentki
M
M
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002951:Citrates / Q000378:metabolism; D002952:Citric Acid Cycle / Q000502:physiology*; D003600:Cytosol / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005947:Glucose / Q000502:physiology*; D066298:In Vitro Techniques; D007328:Insulin / Q000378:metabolism; D007329:Insulin Antagonists / Q000494:pharmacology; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism*; D010648:Phenylacetates / Q000494:pharmacology; D011766:Pyruvate Carboxylase / Q000037:antagonists & inhibitors; D011773:Pyruvates / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D015398:Signal Transduction / Q000502:physiology
D002951:Citrates; D004791:Enzyme Inhibitors; D007328:Insulin; D007329:Insulin Antagonists; D010648:Phenylacetates; D011773:Pyruvates; D011766:Pyruvate Carboxylase; C025136:phenylacetic acid; D005947:Glucose; D002118:Calcium