As a first approach, we compared the effect of OT on spontaneous and miniature EPSCs/IPSCs. Whereas miniature currents (i.e. recorded in the presence of TTX) reflect only the activity of individual release sites, spontaneous synaptic currents allow action potentials and network activity-driven neurotransmitter release. TGOT did not change the frequency of occurrence of GABA
A-R-mediated miniature IPSCs (Figure
2B) but significantly increased AMPA-R-mediated synaptic transmission in 50% of the recorded lamina II neurons (Figure
2A). This suggested the presence of functional OT receptors on the presynaptic terminals of a subpopulation of glutamatergic neurons and the absence of such receptors on the synaptic endings of GABAergic dorsal horn interneurons. Although c-Fos expression was not observed in GAD positive neurons following PVN stimulation (Figure
4), we can not fully exclude that OT receptors are expressed by GABAergic interneurones. Should this be the case, we can however conclude that their activation is certainly not sufficient to directly increase the release probability of GABA (i.e. as seen in miniature transmission experiments; Figure
2) or to induce a significant action potential discharge in these neurons (see Figure
5 and related results in the text). These results are in agreement with those reported previously on cultured laminae I-III dorsal horn neurons [
25]. A major difference observed in slices compared to dorsal horn primary cultures concerned the recruitment by OT of a population of AMPA-R mEPSCs with slow rise and decay kinetics (Figure
3). These mEPSCs were rare under control conditions (indicated by the histogram skewness in figure
3A) but were clearly seen after OT receptor stimulation. Their slow kinetics might indicate that they originate at synapses distant from the neuronal cell body, possibly on distal dendrites. In culture, dorsal horn neurons possess simpler dendritic trees and synapses normally impinging on distal dendrites might be established on the cell body and/or proximal dendrites. The existence of silent synapses due to the absence of functional postsynaptic AMPA-R has been previously shown in the spinal cord, in normal and pathological situations [
26,
27]. Our results indicate for the first time that a pool of "presynaptically silent" synapses, which can be recruited or turned on functionally by a neuromodulator such as OT (Figure
3), might exist in the dorsal horn of the spinal cord. Although we cannot completely exclude a postsynaptic locus for the expression of this phenomenon [
28,
29], the fast onset and reversibility TGOT effect on glutamatergic transmission argue rather in favor of the unmasking (i.e. by increasing the release probability) of presynaptically silent or whispering synapses at distal dendrites [
29].