Mediation by calcium/calmodulin-dependent protein kinase II of suppression of GABAA receptors by NMDA

Abstract

Using nystatin-perforated whole-cell recording configuration, the modulatory effect of N-methyl-D-aspartate (NMDA) on gamma-aminobutyric acid (GABA)-activated whole-cell currents was investigated in neurons freshly dissociated from the rat sacral dorsal commissural nucleus (SDCN). The results showed that: (i) NMDA suppressed GABA-and muscimol (Mus)-activated currents (I(gaba) and I(Mus)), respectively in the Mg(2+)-free external solution containing 1 mumol/L glycine at a holding potential (V ( H )) of -40 mV in SDCN neurons. The selective NMDA receptor antagonist, D-2-amino-5-phosphonovaleric acid (APV, 100 gammamol/L), inhibited the NMDA-evoked currents and blocked the NMDA-induced suppression of I(gaba); (ii) when the neurons were incubated in a Ca(2+)-free bath or pre-loaded with a membrane-permeable Ca(2+) chelator, BAPTA AM (10 mumol/L), the inhibitory effect of NMDA on I(GAba) disappeared. Cd(2+) (10 mumol/L) or La(3+) (30 mumol/L), the non-selective blockers of voltage-dependent calcium channels, did not affect the suppression of I(gaba) by NMDA application; (iii) the suppression of I(GAba) by NMDA was inhibited by KN-62, a calcium/calmodulin-dependent protein kinase II (CaMKII) inhibitor. These results indicated that the inhibition of GABA response by NMDA is Ca(2+)-dependent and CaMKII is involved in the process of the Ca(2+)-dependent inhibition.