The action of excitatory amino acids on chick spinal cord neurones in culture.
1. Membrane currents evoked by N-methyl-D-aspartate (NMDA), L-aspartate, L-glutamate, quisqualate and kainate were studied in cultured neurones from the embryonic chick spinal cord by the patch-clamp technique and by employing a quasi-step microperfusion technique. 2. Application of NMDA, aspartate, glutamate and quisqualate induced currents which exhibited an initial peak which declined to a plateau level with a time constant of 2 s and then remained constant or slowly decreased. The discontinuation of the application was followed by an after-current. The individual components of the responses were insensitive to TTX (2 X 10(-6) M) and were present in neurones which did not exhibit any sign of synaptic activity. The responses induced by kainate were monophasic and declined slowly during long-lasting application. 3. The responses induced by NMDA, aspartate and glutamate were voltage dependent, while those induced by kainate were linear between -80 and +80 mV. The equilibrium potential for all components of the responses to all excitatory amino acids was close to zero. 4. From dose-response curves the half-maximum effective dose (ED50) for glutamate and kainate was 3 X 10(-5) and 2 X 10(-4) M respectively. The Hill coefficients for the glutamate and the kainate were calculated to be 1.8 +/- 0.1 (n = 4) and 1.9 +/- 0.5 (n = 4) respectively. Thus two molecules may be interacting with each of the receptor-activated ion channels. 5. Interaction between kainate and quisqualate or kainate and NMDA was studied at both negative and positive holding potentials. No summation of the responses was found when kainate at concentrations close to those required for evoking the maximum response was applied simultaneously with quisqualate or NMDA. On the contrary, a diminution of the membrane currents was observed. A marked decrease in membrane currents was also observed when glutamate (10(-4) M) was applied simultaneously with aspartate (10(-4) M). 6. Glutamate-activated single-channel currents were recorded in the cell-attached configuration with electrodes filled with glutamate (20 microM) in five neurones and a conductance approximately 50 pS was found. 7. It is suggested that differences in the potency of the different excitatory amino acids as open-channel blockers may be one of the mechanisms which contribute to the diversity in the action of excitatory amino acids and that at least some of the effects of NMDA, aspartate, glutamate, quisqualate and kainate may be mediated by a common receptor-channel complex.
D000200:Action Potentials / Q000187:drug effects; D000596:Amino Acids / Q000494:pharmacology*; D000818:Animals; D002478:Cells, Cultured; D002642:Chick Embryo; D004305:Dose-Response Relationship, Drug; D007473:Ion Channels / Q000187:drug effects; D009474:Neurons / Q000187:drug effects* / Q000502:physiology; D013116:Spinal Cord / Q000502:physiology*; D013779:Tetrodotoxin / Q000494:pharmacology
D000596:Amino Acids; D007473:Ion Channels; D013779:Tetrodotoxin