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Received May 25, 2004
Revised June 3, 2004
Accepted after revision June 22, 2004
1 Centre médical universitaire. Université de Genève.
2 UMR 7519 CNRS/ULP
* To whom correspondence should be addressed. E-mail: trouslard{at}neurochem.u-strasbg.fr.
Using whole cell voltage clamp recordings from lamina X neurones in rat spinal cord slices we investigated the effect of glycine transporter (GlyT) antagonists on both glycinergic inhibitory postsynaptic current (IPSCs) and glutamatergic excitatory post synaptic current (EPSCs). In this study we have used ORG 24598 and ORG 25543, two selective antagonists of the glial GlyT (GlyT1) and neuronal GlyT (GlyT2) respectively. In rats (P12-P16) and in the presence of kynurenic acid, CNQX and bicuculline, ORG 24598 and ORG 25543 applied individually at a concentration of 10 µM, induced a mean inward current of -10/-50 pA at -60 mV and increased significantly the decay time constants of miniature (mIPSCs), spontaneous (sIPSCs) and electrically-evoked glycinergic (eIPSCs) inhibitory postsynaptic currents. ORG 25543, but not ORG 24598, decreased the frequency of mIPSCs and sIPSCs. Substituting extracellular sodium with N-methyl-D-glucamine or superfusing the slice with µM concentration of glycine also increased the decay time constant of glycinergic IPSCs. By contrast, the decay time constant, amplitude and frequency of miniature GABAergic IPSCs recorded in the presence of strychnine were not affected by ORG 24598 and ORG 25543. In the presence of strychnine, bicuculline and 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), we recorded electrically evoked NMDA receptor-mediated EPSCs (eEPSCs). eEPSCs were suppressed by 30 µM D-2-amino-5-phosphonovalerate (APV), an antagonist of the NMDA receptor and by 30 µM dichlorokynurenic acid (DCKA), an antagonist of the glycine site of the NMDA receptor. Glycine (1-5 µM) and D-serine (10 µM) increased the amplitude of eEPSCs whereas L-serine had no effect. ORG 24598 and ORG 25543 increased significantly the amplitude of NMDA receptor-mediated eEPSCs without affecting the amplitude of non NMDA receptor-mediated eEPSCs.
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