title: Increased uncoupling protein-2 levels in beta-cells are associated with impaired glucose-stimulated insulin secretion: mechanism of action.
abstract: In pancreatic beta-cells, glucose metabolism signals insulin secretion by altering the cellular array of messenger molecules. ATP is particularly important, given its role in regulating cation channel activity, exocytosis, and events dependent upon its hydrolysis. Uncoupling protein (UCP)-2 is proposed to catalyze a mitochondrial inner-membrane H(+) leak that bypasses ATP synthase, thereby reducing cellular ATP content. Previously, we showed that overexpression of UCP-2 suppressed glucose-stimulated insulin secretion (GSIS) in isolated islets (1). The aim of this study was to identify downstream consequences of UCP-2 overexpression and to determine whether insufficient insulin secretion in a diabetic model was correlated with increased endogenous UCP-2 expression. In isolated islets from normal rats, the degree to which GSIS was suppressed was inversely correlated with the amount of UCP-2 expression induced. Depolarizing the islets with KCl or inhibiting ATP-dependent K(+) (K(ATP)) channels with glybenclamide elicited similar insulin secretion in control and UCP-2-overexpressing islets. The glucose-stimulated mitochondrial membrane ((m)) hyperpolarization was reduced in beta-cells overexpressing UCP-2. ATP content of UCP-2-induced islets was reduced by 50%, and there was no change in the efflux of Rb(+) at high versus low glucose concentrations, suggesting that low ATP led to reduced glucose-induced depolarization, thereby causing reduced insulin secretion. Sprague-Dawley rats fed a diet with 40% fat for 3 weeks were glucose intolerant, and in vitro insulin secretion at high glucose was only increased 8.5-fold over basal, compared with 28-fold in control rats. Islet UCP-2 mRNA expression was increased twofold. These studies provide further strong evidence that UCP-2 is an important negative regulator of beta-cell insulin secretion and demonstrate that reduced (m) and increased activity of K(ATP) channels are mechanisms by which UCP-2-mediated effects are mediated. These studies also raise the possibility that a pathological upregulation of UCP-2 expression in the prediabetic state could contribute to the loss of glucose responsiveness observed in obesity-related type 2 diabetes in humans.
authors.affiliation: Department of Anatomy and Physiology, Atlantic Veterinary College, University of Prince Edward Island, 550 University Avenue, Charlottetown, PEI C1A 4P3, Canada. cchan@upei.ca
authors.forename: C B
authors.initials: CB
authors.lastname: Chan
authors.forename: D
authors.initials: D
authors.lastname: De Leo
authors.forename: J W
authors.initials: JW
authors.lastname: Joseph
authors.forename: T S
authors.initials: TS
authors.lastname: McQuaid
authors.forename: X F
authors.initials: XF
authors.lastname: Ha
authors.forename: F
authors.initials: F
authors.lastname: Xu
authors.forename: R G
authors.initials: RG
authors.lastname: Tsushima
authors.forename: P S
authors.initials: PS
authors.lastname: Pennefather
authors.forename: A M
authors.initials: AM
authors.lastname: Salapatek
authors.forename: M B
authors.initials: MB
authors.lastname: Wheeler
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism / Q000502:physiology; D000818:Animals; D004041:Dietary Fats / Q000008:administration & dosage / Q000494:pharmacology; D004594:Electrophysiology; D005947:Glucose / Q000494:pharmacology*; D006801:Humans; D007328:Insulin / Q000378:metabolism*; D000078790:Insulin Secretion; D007425:Intracellular Membranes / Q000187:drug effects / Q000502:physiology; D007473:Ion Channels; D007515:Islets of Langerhans / Q000187:drug effects* / Q000378:metabolism*; D026901:Membrane Transport Proteins; D008928:Mitochondria / Q000187:drug effects / Q000502:physiology; D024101:Mitochondrial Proteins; D015221:Potassium Channels / Q000378:metabolism; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D012016:Reference Values; D012413:Rubidium / Q000378:metabolism; D000071246:Uncoupling Protein 2
chemical_list: D004041:Dietary Fats; D007328:Insulin; D007473:Ion Channels; D026901:Membrane Transport Proteins; D024101:Mitochondrial Proteins; D015221:Potassium Channels; D011506:Proteins; D012333:RNA, Messenger; C000604389:UCP2 protein, human; C000604391:Ucp2 protein, rat; D000071246:Uncoupling Protein 2; D000255:Adenosine Triphosphate; D005947:Glucose; D012413:Rubidium