Expression of Ca(2+)-ion permeable alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptors in Xenopus oocytes injected with total RNA from human epileptic temporal lobe.
By using the Xenopus oocyte as an expression system, we have performed a series of experiments in order to examine the divalent cation-permeability of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptors from the human epileptic temporal lobe. Xenopus oocytes, injected with total RNA from the epileptic temporal lobe, were tested for expression of receptors by a conventional two electrode voltage-clamp technique. Administration of glutamate and AMPA gave small or no clear current responses, whereas kainate produced large inward non-desensitizing currents. The current responses evoked by kainate were concentration dependent. Experimental data gave a Hill coefficient of 1.06 and an EC50 value of 87 microM. The current to voltage relationship showed an inward rectification and when the concentration of divalent cations were enhanced, there was a shift in the reversal potential from -11 mV (2 mM Ca2+) to 12 mV (60 mM Ba2+). This yielded a pBa2+/pK+ permeability ratio of 1.6 when the constant field equation was used. The amplitude of the currents evoked by 600 microM kainate in solutions containing higher Ba(2+)-ion concentrations was markedly diminished (46% in 10 mM Ba(2+)- and 75% in 60 mM Ba(2+)-solution), when compared to those obtained in normal Ringer's solution, suggesting interactions between different cation species and/or screening of surface charges.
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D004827:Epilepsy / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D007608:Kainic Acid / Q000494:pharmacology; D009865:Oocytes; D012313:RNA / Q000494:pharmacology*; D018091:Receptors, AMPA / Q000235:genetics*; D013702:Temporal Lobe / Q000503:physiopathology*; D014981:Xenopus
D018091:Receptors, AMPA; D012313:RNA; D007608:Kainic Acid; D002118:Calcium