Inhibition of MK801 binding in adult rat brain sections by conantokin-G and conantokin-T.

The functional interactions of conantokins with anatomical sites in rat brain have been assessed through displacement of the non-competitive N-methyl-D-aspartate receptor (NMDAR) antagonist, dizocilpine (MK801). The binding of (+)-3-[125I]-iodo-MK801 (1 nM) to coronal sections from adult rat brain was inhibited in a dose-dependent manner by conantokin-T (con-T) and conantokin-G (con-G). Quantitative densitometry was used to determine IC50 values for conantokin inhibition of [125I]-MK801 binding in the cortex, thalamus and hippocampus. Con-T completely inhibited [125I]-MK801 specific binding in all brain regions at a saturating concentration of 100 microM. Con-G was able to completely displace [125I]-MK801 in the cortex and thalamus, but only inhibited this same binding up to approximately 90% in the hippocampus. Both peptides maintained their inhibitory properties in the presence of 1 mM EDTA, suggesting that divalent cations are not required for their action in this regard. The added presence of spermine (150 microM) resulted in a two-fold increase in [125I]-MK801 binding and a two-fold decrease in the IC50 values for both peptides. The data obtained in this investigation further demonstrate that [125I]-MK801 is a useful probe for the indirect determination of functional NMDAR ligand binding sites in rat brain sections.