Characterization of spontaneous excitatory synaptic currents in newt retinal bipolar cells

The kinetics of glutamate concentration in the synaptic cleft is an important determinant of synaptic function. To elucidate peak concentration of glutamate released from a single vesicle in the cleft, spontaneous excitatory postsynaptic currents (sEPSCs) in Off-bipolar cells from the sliced newt retina were analyzed using whole-cell patch clamp recording and the computer simulation. The sEPSCs were blocked by an AMPA/kainate (KA) antagonist, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX), and prolonged by cyclothiazide. However, an N-methyl-D-aspartate (NMDA) antagonist, D-2-amino-5-phosphonopentanoic acid (D-AP5), was ineffective. These suggest that sEPSCs in Off-bipolar cells are mediated exclusively by AMPA/KA receptors. sEPSCs simulated by a detailed kinetic model of AMPA receptor best approximated the data, when peak glutamate concentration was 10 microM. Therefore, it was concluded that peak concentration of glutamate released from a single vesicle would be elevated to approximately 10 microM at the newt Off-bipolar dendrite.