Calcium chelators are commonly used for fluorescence and electrophysiological studies of neuronal Ca2+ signalling. Recently, they have also been used as neuroprotectants. Since they buffer calcium ions, these agents also modify the same signals which are being studied. These properties may be used to modulate Ca2+ signals such as those involved in synaptic transmission, and may explain their neuroprotective mechanism. To define factors which govern the modulation of synaptic transmission by Ca2+ chelators, we examined their actions on synaptic responses evoked in CA1 neurons of rat hippocampal slices. We used a spectrum of cell-permeant Ca2+ chelators having different structures, Ca(2+)-binding kinetics and Ca2+ affinities, as well as an impermeant, intracellularly perfused chelator salt. Application of the cell-permeant 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetra-acetate acetoxymethyl ester (50 microM) markedly attenuated evoked synaptic responses. This application produced an intracellular chelator accumulation of 79-125 microM, as estimated using 14C-labelled chelator. The actions of a Ca2+ chelator on synaptic responses were dependent on the chelator's Ca2+ affinity, Ca(2+)-binding rate and Ca2+ selectivity, because 1,2-bis(2-amino-5-nitrophenoxy)ethane-N,N,N',N'-tetra-acetate acetoxymethyl ester (a low Ca2+ affinity analogue), ethyleneglycolbis(beta-aminoethyl ether)-N,N,N',N'-tetra-acetate acetoxymethyl ester (a slow buffer with similar Ca2+ affinity to 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetra-acetate) and the selective Zn2+ chelator, tetrakis(2-pyridylmethyl)ethylenediamine, were ineffective. The intrinsic cell membrane properties, including the post-spike train afterhyperpolarization, were not significantly affected by any of the Ca2+ chelators used in this study. Intracellular perfusion of 100-200 microM 1,2-bis-(2-aminophenoxy)ethane-N,N,N',N'-tetra-acetate salt through patch pipettes into postsynaptic cells did not affect synaptic potentials, suggesting a presynaptic action of cell-permeant Ca2+ chelators on transmitter release. Other cell-permeant, fast Ca(2+)-binding chelators reduced synaptic responses according to their Ca2+ affinities, and not their chemical structure: those chelators with Kd values < or = 25 microM attenuated synaptic responses, whereas chelators of lesser affinity did not. These data support the ideas that [Ca2+]i rises to high (micromolar) levels during transmitter release, and that Ca2+ chelators may be used to attenuate excitotoxicity by attenuating excitatory neurotransmission without affecting Ca2+ signalling in the submicromolar [Ca2+]i range.