1 A grease-gap recording technique has been used to investigate the mechanisms underlying the acute potentiation of N-methyl-D-aspartate (NMDA) responses by aminocyclopentane-1S,3R-dicarboxylic acid (IS,3R-ACPD) in area CAI of rat hippocampal slices.2 1S,3R-ACPD (1OM), but not IR,3S-ACPD (1OEM), potentiated submaximal responses to NMDA (dose-ratio of 0.81 ± 0.02 (mean ± s.e.mean); n = 55), but not to a-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA), in a readily reversible manner. Potentiation also occurred in slices treated with 0.2 tLM tetrodotoxin, and in slices perfused with Mg2"-free medium.3 IS,3R-ACPD-induced potentiation was unaffected by the protein kinase inhibitors K-252b (0.1 tLM) and staurosporine (1 gM) and the intracellular Ca2+ store depletor, thapsigargin (10 gM). Coapplication of staurosporine and thapsigargin was also without effect. 4 IS,3R-ACPD-induced potentiation was unaffected by inhibitors of arachidonic acid formation, bromophenacyl bromide (50 tM) and RG80267 (100 gM). Arachidonic acid (10-50 gM) depressed reversibly NMDA-induced responses. The potentiation was unaffected by 8-bromo cyclic AMP (500 jAM) or the PKA inhibitor Rp-adenosine 3,5-cyclic monophosphothioate triethylamine (Rp-cAMPS; 50 jAM). 5 IS,3R-ACPD-induced potentiation was abolished in slices perfused with Ca2+-free medium. The potentiation was also blocked by phorbol-12,13-diacetate (1 gM), in a staurosporine-sensitive manner.6 It is concluded that the potentiation of NMDA responses by IS,3R-ACPD is not mediated by protein kinase A or C, by release of Ca2+ from intracellular stores or by arachidonic acid. It involves a Ca2+-sensitive process and is negatively regulated by protein kinase C.