title: Functional expression and pharmacological characterization of the human EAA4 (GluR6) glutamate receptor: a kainate selective channel subunit.
abstract: A cDNA encoding an ionotropic glutamate receptor subunit protein humEAA4 (GluR6), has been cloned from a human fetal brain library. This cDNA when expressed in COS or HEK-293 cells is associated with high-affinity kainate receptor binding and ion channel formation. We have successfully established cell lines stably expressing humEAA4 in HEK-293 cells This is the first report of the establishment of stable cell lines expressing a glutamate receptor channel. The relative potency of compounds for displacing [3H]-kainate binding to humEAA4 receptors expressed in COS or HEK-293 cells is domoate > kainate > quisqualate > 6-cyano-7-nitroquinoxaline-2,3-dione > L-glutamate = 6,7- dinitroquinoxaline-2,3-dione > dihydrokainate. Applications of kainate, glutamate, and domoate but not AMPA evoked rapidly desensitizing currents in cells expressing homo-oligomeric humEAA4 in a concentration dependent manner. The order of potency was: domoate > kainate > L-glutamate. Although AMPA did not itself activate humEAA4 receptors it did reduce, to a limited extent, kainate-evoked responses. AMPA may therefore be a weak partial agonist for this receptor. To date this effect has not been demonstrated with rat GluR6. It is possible that subtle species differences may exist in the nature of agonist receptor interaction. Kainate evoked currents were attenuated by the quinoxalinediones CNQX and DNQX but not by DAP5. The receptor desensitization was attenuated on application of concanavalin A. Ion-permeability studies indicated that the receptor-linked ion channel is permeable to both Na+ and Ca2+ ions.
authors.affiliation: Allelix Biopharmaceuticals Inc., Mississauga, Ontario, Canada.
authors.forename: K H
authors.initials: KH
authors.lastname: Hoo
authors.forename: S L
authors.initials: SL
authors.lastname: Nutt
authors.forename: E J
authors.initials: EJ
authors.lastname: Fletcher
authors.forename: C E
authors.initials: CE
authors.lastname: Elliott
authors.forename: B
authors.initials: B
authors.lastname: Korczak
authors.forename: R M
authors.initials: RM
authors.lastname: Deverill
authors.forename: V
authors.initials: V
authors.lastname: Rampersad
authors.forename: R P
authors.initials: RP
authors.lastname: Fantaske
authors.forename: R K
authors.initials: RK
authors.lastname: Kamboj
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001667:Binding, Competitive; D001921:Brain / Q000378:metabolism; D002460:Cell Line; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D005333:Fetus; D015723:Gene Library; D006801:Humans; D007473:Ion Channels / Q000096:biosynthesis / Q000502:physiology; D007608:Kainic Acid / Q000378:metabolism / Q000494:pharmacology; D007668:Kidney; D007700:Kinetics; D046911:Macromolecular Substances; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D018408:Patch-Clamp Techniques; D018092:Kainic Acid Receptors / Q000096:biosynthesis* / Q000502:physiology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014162:Transfection
chemical_list: D007473:Ion Channels; D007608:Kainic Acid; D046911:Macromolecular Substances; D015345:Oligonucleotide Probes; D018092:Kainic Acid Receptors; D011994:Recombinant Proteins