The K+-evoked release of dynorphin A(1-8)-like immunoreactivity from guinea pig hippocampal mossy fiber synaptosomes was inhibited 53% by L(+)-2-amino-4-phosphonobutyrate (L(+)APB, 300 microM), a glutamate analogue. Equimolar L(+)APB also inhibited the Ca2+-dependent component of endogenous L-glutamate release from these mossy fiber synaptosomes by 40%. The K+-evoked release of both glutamate and dynorphin A(1-8) from rat hippocampal mossy fiber synaptosomes were unaffected by L(+)APB. It is proposed that L(+)APB selectively suppresses the excitatory mossy fiber synaptic inhibiting the Ca2+-dependent release of glutamate and dynorphin A(1-8) from guinea pig but not rat hippocampal mossy fiber terminals.