Recent anatomical evidence that inhibitory interneurones receive approximately 10 times more synapses from mossy fibres than do principal neurones (Acsády et al. 1998) has led to the re-examination of the extent to which interneurones are involved in CA3 network excitability. Although many of the anatomical and physiological properties of mossy fibre-CA3 interneurone synapses have been previously described (Acsády et al. 1998; Tóth et al. 2000), an investigation into the quantal nature of transmission at this synapse has not yet been conducted. Here, we employed variance-mean (VM) analysis to compare the release probability, quantal size (q) and number of release sites (n) at mossy fibre target neurones in CA3. At six of seven interneurone synapses in which a high concentration of Ca2+ was experimentally imposed, the variance-mean relationship could be approximated by a parabola. Estimates of n were 1-2, and the weighted release probability in normal Ca2+ conditions ranged from 0.34 to 0.51. At pyramidal cell synapses, the variance-mean relationship approximated a linear relationship, suggesting that release probability was significantly lower. The weighted quantal amplitude was similar at interneurone synapses and pyramidal cell synapses, although the variability in quantal amplitude was larger at interneurone synapses. Mossy fibre transmission at CA3 interneurone synapses can be explained by a lower number of release sites, a broader range of release probabilities, and larger range of quantal amplitudes than at CA3 pyramidal synapses. Finally, quantal events on to interneurones elicited spike transmission, owing in part to the more depolarized membrane potential than pyramidal cells. These results suggest that although mossy fibre synapses on to pyramidal cells are associated with a larger number of release sites per synapse, the higher connectivity, higher initial release probability, and larger relative impact per quantum on to CA3 interneurones generate strong feedforward inhibition at physiological firing frequencies of dentate granule cells. Given the central role of CA3 interneurones in mossy fibre synaptic transmission, these details of mossy fibre synaptic transmission should provide insight into CA3 network dynamics under both physiological and pathophysiological circumstances.