title: N-acetylaspartylglutamate stimulates metabotropic glutamate receptor 3 to regulate expression of the GABA(A) alpha6 subunit in cerebellar granule cells.
abstract: We have shown that the vertebrate neuropeptide N-acetylaspartylglutamate (NAAG) meets the criteria for a neurotransmitter, including function as a selective metabotropic glutamate receptor (mGluR) 3 agonist. Short-term treatment of cerebellar granule cells with NAAG (30 microM) results in the transient increase in content of GABA(A) alpha6 subunit mRNA. Using quantitative PCR, this increase was determined to be up to 170% of control values. Similar effects are seen following treatment with trans-1-aminocyclopentane-1,3-dicarboxylate and glutamate and are blocked by the mGluR antagonists (2S,3S,4S)-2-methyl-2-(carboxycyclopropyl) glycine and (2S)-alpha-ethylglutamic acid. The effect is pertussis toxin-sensitive. The increase in alpha6 subunit mRNA level can be simulated by activation of other receptors negatively linked to adenylate cyclase activity, such as adenosine A1, alpha2-adrenergic, muscarinic, and GABA(B) receptors. Forskolin stimulation of cyclic AMP (cAMP) levels abolished the effect of NAAG. The change in alpha6 levels induced by 30 microM NAAG can be inhibited in a dose-dependent manner by simultaneous application of increasing doses of the beta-adrenergic receptor agonist isoproterenol. The increase in alpha6 mRNA content is followed by a fourfold increase in alpha6 protein level 6 h posttreatment. Under voltage-clamped conditions, NAAG-treated granule cells demonstrate an increase in the furosemide-induced inhibition of GABA-gated currents in a concentration-dependent manner, indicating an increase in functional alpha6-containing GABA(A) receptors. These data support the hypothesis that NAAG, acting through mGluR3, regulates expression of the GABA(A) alpha6 subunit via a cAMP-mediated pathway and that cAMP-coupled receptors for other neurotransmitters may similarly influence GABA(A) receptor subunit composition.
authors.affiliation: Department of Biology, Georgetown University, Washington, D.C. 20057-5662, U.S.A.
authors.forename: S
authors.initials: S
authors.lastname: Ghose
authors.forename: B
authors.initials: B
authors.lastname: Wroblewska
authors.forename: L
authors.initials: L
authors.lastname: Corsi
authors.forename: D R
authors.initials: DR
authors.lastname: Grayson
authors.forename: A L
authors.initials: AL
authors.lastname: De Blas
authors.forename: S
authors.initials: S
authors.lastname: Vicini
authors.forename: J H
authors.initials: JH
authors.lastname: Neale
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000378:metabolism* / Q000502:physiology; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000502:physiology; D004151:Dipeptides / Q000494:pharmacology*; D004553:Electric Conductivity; D005665:Furosemide / Q000494:pharmacology; D015640:Ion Channel Gating / Q000502:physiology; D009474:Neurons / Q000378:metabolism* / Q000502:physiology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011963:Receptors, GABA-A / Q000235:genetics / Q000378:metabolism*; D018094:Receptors, Metabotropic Glutamate / Q000502:physiology*; D014158:Transcription, Genetic; D005680:gamma-Aminobutyric Acid / Q000502:physiology
chemical_list: D004151:Dipeptides; D012333:RNA, Messenger; D011963:Receptors, GABA-A; D018094:Receptors, Metabotropic Glutamate; C102718:metabotropic glutamate receptor 3; D005576:Colforsin; C027172:isospaglumic acid; D005680:gamma-Aminobutyric Acid; D005665:Furosemide; D000242:Cyclic AMP