The interactions of guanine nucleotides, and particularly GTP, with the [3H]-alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate (AMPA) and [3H]-kainate (KA) binding sites present on brain membranes was studied, using the ligand binding methodology and Scatchard analysis, in order to establish the competitive/non competitive nature of the interaction and determine whether guanine nucleotides, KA and AMPA share common binding sites. GTP was found to block [3H]-AMPA and [3H]-KA binding to rat cortical membranes with IC50 values of 0.4 mM and 1 mM respectively and the [3H] KA-binding to chick cerebellar membranes with a IC50 value of 20 microM. Scatchard analysis of [3H]-KA binding performed in the absence or presence of 1 mM GTP or 0.25 mM AMPA reveals that the high affinity [3H]-KA binding component is not affected by GTP but blocked in a non competitive fashion by AMPA while the low affinity [3H]-KA binding component is not affected by AMPA but blocked by GTP. Scatchard analysis of [3H]-KA binding to chick cerebellar membranes performed in the absence or presence of 33 microM GTP reveals a single binding site blocked in a competitive fashion by GTP. Scatchard analysis of [3H]-AMPA binding performed in the absence or presence of 0.5 mM GTP or 30 microM KA reveals that the high affinity [3H]-AMPA binding component is affected in a non competitive fashion by both GTP and KA while the low affinity [3H] AMPA binding component is affected in a competitive fashion by both GTP and KA.(ABSTRACT TRUNCATED AT 250 WORDS)