A large number of septal neurons express alpha-amino-3-hydroxy-5-methyl-4-isoxazoleproprionate (AMPA)-type excitatory glutamate receptors. It has been demonstrated that in the mediolateral part of the rat lateral septum, calbindin-containing neurons are heavily innervated by hypothalamic, enkephalinergic fibers forming exclusively asymmetric synaptic contacts. This connection was suggested to be excitatory. In order to further elucidate this hypothesis, the aim of the present study was to determine whether these enkephalinoceptive neurons express GluR1 and GluR2/3 AMPA receptor subunits. Correlated light and electron microscopic analysis, using single immunostaining for GluR1 and GluR2/3, and double immunostaining for Leu-enkephalin and GluR1 or GluR2/3, was performed on vibratome sections of the rat lateral septal area. The studies revealed that while GluR1 is mainly associated with dendritic and somatic spines, GluR2/3 is mostly present in the perisomatic area. Leu-enkephalin boutons establish asymmetric synaptic contacts at the level of the soma and initial dendrites of both of these cells. A semiquantitative analysis showed that these enkephalin-targeted cells represent 50% of the total number of both GluR1 and GluR2/3-containing lateral septal neurons. These results suggest that: (1) AMPA receptor-expressing neurons appear to be the exclusive recipient of hypothalamic Leu-enkephalin boutons; (2) these enkephalinoceptive neurons contain both GluR1 and GluR2/3 AMPA receptor subunits; however, (3) only the GluR2/3 subtype, located in the perisomatic area, may be associated with Leu-enkephalin-containing inputs.