title: Ablation of AMP-activated protein kinase alpha1 and alpha2 from mouse pancreatic beta cells and RIP2.Cre neurons suppresses insulin release in vivo.
abstract: OBJECTIVE
  AMP-activated protein kinase (AMPK) is an evolutionarily conserved enzyme and a target of glucose-lowering agents, including metformin. However, the precise role or roles of the enzyme in controlling insulin secretion remain uncertain.
  
  
  METHODS
  The catalytic alpha1 and alpha2 subunits of AMPK were ablated selectively in mouse pancreatic beta cells and hypothalamic neurons by breeding Ampkalpha1 [also known as Prkaa1]-knockout mice, bearing floxed Ampkalpha2 [also known as Prkaa2] alleles (Ampkalpha1 ( -/- ),alpha2( fl/fl ),), with mice expressing Cre recombinase under the rat insulin promoter (RIP2). RIP2 was used to express constitutively activated AMPK selectively in beta cells in transgenic mice. Food intake, body weight and urinary catecholamines were measured using metabolic cages. Glucose and insulin tolerance were determined after intraperitoneal injection. Beta cell mass and morphology were analysed by optical projection tomography and confocal immunofluorescence microscopy, respectively. Granule docking, insulin secretion, membrane potential and intracellular free Ca(2+) were measured with standard techniques.
  
  
  RESULTS
  Trigenic Ampkalpha1 ( -/- ),alpha2( fl/fl ) expressing Cre recombinase and lacking both AMPKalpha subunits in the beta cell, displayed normal body weight and increased insulin sensitivity, but were profoundly insulin-deficient. Secreted catecholamine levels were unchanged. Total beta cell mass was unaltered, while mean islet and beta cell volume were reduced. AMPK-deficient beta cells displayed normal glucose-induced changes in membrane potential and intracellular free Ca(2+), while granule docking and insulin secretion were enhanced. Conversely, betaAMPK transgenic mice were glucose-intolerant and displayed defective insulin secretion.
  
  
  CONCLUSIONS
  Inhibition of AMPK activity within the beta cell is necessary, but not sufficient for stimulation of insulin secretion by glucose to occur. AMPK activation in extrapancreatic RIP2.Cre-expressing cells might also influence insulin secretion in vivo.
authors.affiliation: Section of Cell Biology, Division of Diabetes, Endocrinology and Metabolism, Department of Medicine, Imperial College London, Sir Alexander Fleming Building, Exhibition Road, South Kensington, London, SW7 2AZ, UK.
authors.forename: G
authors.initials: G
authors.lastname: Sun
authors.forename: A I
authors.initials: AI
authors.lastname: Tarasov
authors.forename: J
authors.initials: J
authors.lastname: McGinty
authors.forename: A
authors.initials: A
authors.lastname: McDonald
authors.forename: G
authors.initials: G
authors.lastname: da Silva Xavier
authors.forename: T
authors.initials: T
authors.lastname: Gorman
authors.forename: A
authors.initials: A
authors.lastname: Marley
authors.forename: P M
authors.initials: PM
authors.lastname: French
authors.forename: H
authors.initials: H
authors.lastname: Parker
authors.forename: F
authors.initials: F
authors.lastname: Gribble
authors.forename: F
authors.initials: F
authors.lastname: Reimann
authors.forename: O
authors.initials: O
authors.lastname: Prendiville
authors.forename: R
authors.initials: R
authors.lastname: Carzaniga
authors.forename: B
authors.initials: B
authors.lastname: Viollet
authors.forename: I
authors.initials: I
authors.lastname: Leclerc
authors.forename: G A
authors.initials: GA
authors.lastname: Rutter
mesh_terms: D055372:AMP-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*; D000704:Analysis of Variance; D000818:Animals; D001786:Blood Glucose / Q000378:metabolism; D001835:Body Weight / Q000235:genetics; D004041:Dietary Fats; D004435:Eating / Q000235:genetics; D004594:Electrophysiology; D005455:Fluorescent Antibody Technique; D005951:Glucose Tolerance Test; D006943:Hyperglycemia / Q000235:genetics / Q000378:metabolism; D007031:Hypothalamus / Q000378:metabolism*; D007328:Insulin / Q000235:genetics / Q000378:metabolism*; D000078790:Insulin Secretion; D050417:Insulin-Secreting Cells / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D009474:Neurons / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D051381:Rats
chemical_list: D001786:Blood Glucose; D004041:Dietary Fats; D007328:Insulin; D055372:AMP-Activated Protein Kinases