title: Excitatory amino acid-induced currents in rat septal cholinergic neurons in culture.
abstract: Whole-cell voltage-clamp recordings were used to study excitatory amino acid-induced currents in neurons isolated from the septum of fetal rat brains. The neurons were cultured for more than four weeks on a feeder layer composed of glial cells obtained from the septal region. Septal neurons were either fusiform, triangular or multipolar and 83% of cells showed acetylcholinesterase activity. L-Glutamate, kainate, quisqualate and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) applied by local perfusion produced inward currents (Iglu, Ikai, Iquis and IAMPA, respectively) at -44mV which increased in amplitude with increasing concentration of agonist; they desensitized when induced at higher concentrations except for the Ikai. The EC50s for the peak Ikai and sustained Iglu, Iquis and IAMPA were 55, 13, 0.39 and 3.5 microM, respectively. 6-Cyano-7-nitroquinoxaline-2,3-dione (CNQX) depressed Ikai and IAMPA evoked at a concentration of 10 microM (IC50s: 0.58 and 0.84 microM, respectively). Schild analysis for the CNQX action on Ikai gave a dissociation constant of 0.27 microM for CNQX. n-Methyl-D-aspartate (NMDA) (with glycine, 3 microM) produced an inward current (INMDA) at -44 mV whose peak amplitude enhanced with increased concentrations (EC50 = 32 microM). INMDA was potentiated by glycine (EC50 = 0.15 microM) and inhibited by D-2-amino-5-phosphovalerate (IC50 = 9.9 microM for INMDA evoked at a concentration of 50 microM). MK-801 (0.1-10 microM) inhibited INMDA in a dose- and use-dependent manner. INMDA was (0.1-10 microM) inhibited INMDA in a dose- and use-dependent manner. INMDA was potentiated by spermine (EC50 = 247 microM; 91% increase at 1mM) in a manner independent of holding potential (VH). INMDA was inhibited by Mg2+ and Zn2+ (IC50 = 673 and 39 microM, respectively, at -44 mV) in a manner dependent on VH; the magnitudes of a depolarization required for an e-fold increase in their IC50s in a range of -64 to -24 mV were 16 and 22 mV, respectively. The action of Zn2+ was independent of VH > -24 mV. Current-voltage relations for Ikai, Iquis and IAMPA exhibited outward rectification, while that of INMDA showed a region of negative conductance at VH < -30 mV, which disappeared in a Mg(2+)-free solution. Reversal potentials for Ikai, Iquis, IAMPA and INMDA were close to 0 mV, indicating the involvement of non-specific cation channels. Increasing extracellular Ca2+ concentration from 2.4 to 30 mM did not affect the Ikai and Iquis, reversal potential showing negligible Ca2+ component, but shifted INMDA reversal potential to a more positive potential, yielding a ratio of Ca2+ permeability to that of monovalent cation to be 13. Cholinergic septal neurons in culture express non-NMDA-(AMPA/kainate-) and NMDA-type of glutamate receptor channels. Their properties were quantitatively similar to those of glutamate receptor channels on other types of neurons in the brain except for the actions of endogenous neuromodulators (Mg2+, Zn2+ and spermine) on NMDA receptor channels. It is suggested that NMDA receptor channels on different types of neurons may play a distinct role depending on a difference in the actions of these neuromodulators.
authors.affiliation: Department of Physiology, Saga Medical School, Japan.
authors.forename: E
authors.initials: E
authors.lastname: Kumamoto
authors.forename: Y
authors.initials: Y
authors.lastname: Murata
mesh_terms: D000200:Action Potentials / Q000187:drug effects* / Q000502:physiology; D000818:Animals; D002478:Cells, Cultured / Q000187:drug effects; D002799:Cholinergic Fibers / Q000187:drug effects*; D018846:Excitatory Amino Acids / Q000494:pharmacology*; D007608:Kainic Acid / Q000494:pharmacology; D007700:Kinetics; D016202:N-Methylaspartate / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D018408:Patch-Clamp Techniques; D051381:Rats; D017207:Rats, Sprague-Dawley; D012686:Septal Nuclei / Q000187:drug effects / Q000502:physiology*; D015032:Zinc / Q000494:pharmacology; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid / Q000494:pharmacology
chemical_list: D018846:Excitatory Amino Acids; D016202:N-Methylaspartate; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid; D015032:Zinc; D007608:Kainic Acid