title: AMPA and kainate receptors mediate mutually exclusive effects on GABA(A) receptor expression in cultured mouse cerebellar granule neurones.
abstract: Studies on animal models of epilepsy and cerebellar ataxia, e.g., stargazer mice (stg) have identified changes in the GABAergic properties of neurones associated with the affected brain loci. Whether these changes contribute to or constitute homeostatic adaptations to a state of altered neuronal excitability is as yet unknown. Using cultured cerebellar granule neurones from control [+/+; alpha-amino-3-hydroxyl-5-methyl-4-isoxazolepropionate receptor (AMPAR)-competent, Kainate receptor (KAR)-competent] and stg (AMPAR-incompetent, KAR-competent), we investigated whether non-NMDA receptor (NMDAR) activity regulates GABA(A) receptor (GABAR) expression. Neurones were maintained in 5 mmol/L KCl-containing basal media or depolarizing media containing either 25 mmol/L KCl or the non-NMDAR agonist kainic acid (KA) (100 micromol/L). KCl- and KA-mediated depolarization down-regulated GABAR alpha1, alpha6 and beta2, but up-regulated alpha4, beta3 and delta subunits in +/+ neurones. The KCl-evoked but not KA-evoked effects were reciprocated in stg neurones compatible with AMPAR-regulation of GABAR expression. Conversely, GABAR gamma2 expression was insensitive to KCl-mediated depolarization, but was down-regulated by KA-treatment in a 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX)-reversible manner in +/+ and stg neurones compatible with a KAR-mediated response. KA-mediated up-regulation of GABAR alpha4, beta3 and delta was inhibited by L-type voltage-gated calcium channel (L-VGCC) blockers and the Ca2+/calmodulin-dependent protein kinase inhibitor, 4-[(2S)-2-[(5-isoquinolinylsulfonyl)methylamino]-3-oxo-3-(4-phenyl-1-piperazinyl)propyl] phenyl isoquinoline sulfonic acid ester (KN-62). Up-regulation of GABAR alpha4 and beta3 was also prevented by calcineurin (CaN) inhibitors, FK506 and cyclosporin A. Down-regulation of GABAR alpha1, alpha6 and beta2 was independent of L-VGCC activity, but was prevented by inhibitors of CaN. Thus, we provide evidence that a KAR-mediated and at least three mutually exclusive AMPAR-mediated signalling mechanisms regulate neuronal GABAR expression.
authors.affiliation: Centre for Integrative Neurosciences, School of Biological and Biomedical Sciences, University of Durham, Durham, UK. H.L.Payne@durham.ac.uk
authors.forename: Helen L
authors.initials: HL
authors.lastname: Payne
authors.forename: Jane H
authors.initials: JH
authors.lastname: Ives
authors.forename: Werner
authors.initials: W
authors.lastname: Sieghart
authors.forename: Christopher L
authors.initials: CL
authors.lastname: Thompson
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001386:Azides / Q000493:pharmacokinetics; D001569:Benzodiazepines / Q000493:pharmacokinetics; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology*; D018846:Excitatory Amino Acids / Q000494:pharmacology; D018755:GABA Agonists / Q000493:pharmacokinetics; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D007608:Kainic Acid / Q000494:pharmacology; D051379:Mice; D008818:Mice, Neurologic Mutants; D008954:Models, Biological; D009118:Muscimol / Q000493:pharmacokinetics; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D011189:Potassium Chloride / Q000494:pharmacology; D021122:Protein Subunits / Q000235:genetics / Q000378:metabolism; D011869:Radioligand Assay / Q000379:methods; D018091:Receptors, AMPA / Q000502:physiology*; D011963:Receptors, GABA-A / Q000378:metabolism*; D018092:Kainic Acid Receptors / Q000502:physiology*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D014316:Tritium / Q000493:pharmacokinetics
chemical_list: D001386:Azides; D001569:Benzodiazepines; D018846:Excitatory Amino Acids; D018755:GABA Agonists; D007608:Kainic Acid; D009118:Muscimol; D011189:Potassium Chloride; D021122:Protein Subunits; D018091:Receptors, AMPA; D011963:Receptors, GABA-A; D018092:Kainic Acid Receptors; D014316:Tritium; C042957:Ro 15-4513