title: Regulation of glucose transporter 3 surface expression by the AMP-activated protein kinase mediates tolerance to glutamate excitation in neurons.
abstract: Ischemic and excitotoxic events within the brain result in rapid and often unfavorable depletions in neuronal energy levels. Here, we investigated the signaling pathways activated in response to the energetic stress created by transient glutamate excitation in cerebellar granule neurons. We characterized a glucose dependent hyperpolarization of the mitochondrial membrane potential (Delta psi(m)) in the majority of neurons after transient glutamate excitation. Expression levels of the primary neuronal glucose transporters (GLUTs) isoforms 1, 3, 4, and 8 were found to be unaltered within a 24 h period after excitation. However, a significant increase only in GLUT3 surface expression was identified 30 min after excitation, with this high surface expression remaining significantly above control levels in many neurons for up to 4 h. Glutamate excitation induced a rapid alteration in the AMP:ATP ratio that was associated with the activation of the AMP-activated protein kinase (AMPK). Interestingly, pharmacological activation of AMPK with AICAR (5-aminoimidazole-4-carboxamide riboside) alone also increased GLUT3 surface expression, with a hyperpolarization of Delta psi(m) evident in many neurons. Notably, inhibition of the CaMKK (calmodulin-dependent protein kinase kinase) had little affect on GLUT translocation, whereas the inhibition or knockdown of AMPK (compound C, siRNA) activity prevented GLUT3 translocation to the cell surface after glutamate excitation. Furthermore, gene silencing of GLUT3 eradicated the increase in Delta psi(m) associated with transient glutamate excitation and potently sensitized neurons to excitotoxicity. In summary, our data suggest that the activation of AMPK and its regulation of cell surface GLUT3 expression is critical in mediating neuronal tolerance to excitotoxicity.
authors.affiliation: Department of Physiology and Medical Physics and Royal College of Surgeons in Ireland Neuroscience Research Centre, Dublin 2, Ireland.
authors.forename: Petronela
authors.initials: P
authors.lastname: Weisová
authors.forename: Caoimhín G
authors.initials: CG
authors.lastname: Concannon
authors.forename: Marc
authors.initials: M
authors.lastname: Devocelle
authors.forename: Jochen H M
authors.initials: JH
authors.lastname: Prehn
authors.forename: Manus W
authors.initials: MW
authors.lastname: Ward
mesh_terms: D000620:Aminoimidazole Carboxamide / Q000031:analogs & derivatives; D006690:Bisbenzimidazole; D015153:Blotting, Western; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000502:physiology; D002462:Cell Membrane / Q000502:physiology; D002531:Cerebellum / Q000166:cytology / Q000502:physiology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000235:genetics / Q000502:physiology*; D018846:Excitatory Amino Acids / Q000633:toxicity*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation / Q000235:genetics / Q000502:physiology*; D005947:Glucose / Q000502:physiology; D051274:Glucose Transporter Type 3 / Q000096:biosynthesis* / Q000235:genetics; D018698:Glutamic Acid / Q000633:toxicity*; D006801:Humans; D018613:Microscopy, Confocal; D008928:Mitochondria / Q000502:physiology / Q000648:ultrastructure; D009474:Neurons / Q000187:drug effects*; D034741:RNA, Small Interfering; D011956:Receptors, Cell Surface / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012265:Ribonucleotides / Q000502:physiology
chemical_list: D018846:Excitatory Amino Acids; D051274:Glucose Transporter Type 3; D034741:RNA, Small Interfering; D011956:Receptors, Cell Surface; D012265:Ribonucleotides; C495687:SLC2A3 protein, human; D000620:Aminoimidazole Carboxamide; D018698:Glutamic Acid; D017868:Cyclic AMP-Dependent Protein Kinases; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; C031143:AICA ribonucleotide; D005947:Glucose; D006690:Bisbenzimidazole