The acidic amino acid antagonist D,L-2-amino-4-phosphonobutyrate (DL-APB) is a potent blocker of synaptic transmission at guinea pig but not rat mossy fiber-CA3 synapses in hippocampal slices. The L-isomer of APB is responsible for the potent inhibition at the guinea pig synapse. The L-APB analogue L-serine-O-phosphate (L-SOP) also is more potent against the guinea pig response. These differences may reflect a difference in a synaptic acidic amino acid receptor in these two species. Other acidic amino acid antagonists are less potent than APB or L-SOP and do not discriminate between the mossy fiber responses in the two species.