Patch clamp analysis of excitatory synapses in mammalian spinal cord slices

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Abstract
Excitatory synaptic transmission to visually identified α-moto neurones was studied in thin slice preparations of the neonatal rat spinal cord. Excitatory postsynaptic currents (EPSCs) elicited by stimulation of intraspinal presynaptic fibres were recorded using the whole-cell patch clamp technique, following blockade of inhibitory transmission by bath application of strychnine and bicuculline. The EPSCs could be separated pharmacologically into N-methyl-d-aspartate- (NMDA) and non-NMDA-receptor-mediated components, where the contribution of the NMDA-mediated component was significant only at holding potentials more positive than −50 mV. Graded stimulation of intraspinal fibres showed that the NMDA- and the non-NMDA-mediated EPSCs were evoked by activation of presynaptic fibres with similar sensitivities to the stimulation intensity, suggesting that the same presynaptic fibres released the excitatory amino-acid (EAA) activating the two sub-sets of receptors. Studies of the amplitude fluctuations of EPSCs elicited by stimulation of a presumed single fibre revealed similar proportions of transmission failures and similar distributions of both the NMDA- and the non-NMDA-mediated components. These similarities suggest that the EAA transmitter activating the two sub-types of receptors is released from the same set of synaptic boutons and that the receptors are therefore post-synaptically co-localized. In addition the gamma aminobutyric acidB (GABAB) receptor agonist l-baclofen, which is known to decrease transmitter release, changed the amplitude distributions of non-NMDA- and NMDA-receptor-mediated EPSCs into unimodal distributions without affecting the amplitude of the presumed unitary event. The similarity between the transmitter release profiles of the two EAA components further supports the notion of postsynaptic receptor co-localization.