Pharmacological characterization, anatomical distribution and sex differences of the non-NMDA excitatory amino acid receptors in the quail brain as identified by CNQX binding

Abstract

The distribution of non- N-methyl- d-aspartate binding sites was studied in coronal and sagittal sections through the brain of adult Japanese quail by quantitative autoradiography, using tritiated 6-cyano-7-nitroquinoxaline-2,3-dione as a radioligand. Saturation binding experiments were, in addition, carried out in areas showing high levels of binding (cerebellar molecular layer, nucleus anterior medialis and nucleus infundibularis) and demonstrated that the binding of tritiated ligand was specific and saturable. Competition studies with α-amino-3-hydroxy-methyl-4-isoxazole propionic acid and kainic acid indicated that kainic acid strongly inhibited ligand binding in all brain areas. α-Amino-3-hydroxy-methyl-4-isoxazole propionic acid was only a weak inhibitor in the hypothalamic nuclei whereas in the cerebellar molecular layer both high and low affinity inhibitions were detected. The highest binding levels of tritiated ligand were observed in the molecular layer of the cerebellum. Very high levels of binding were detected in various preoptic/hypothalamic sites including the nucleus suprachiasmaticus pars medialis, nucleus anterior medialis hypothalami, nucleus infundibularis, nucleus mammillaris medialis, nucleus posteromediale hypothalami and nucleus hypothalami ventromedialis. High levels of binding were also detected in the bulbus olfactorius, bed nucleus commissuralis anterior, bed nucleus commissuralis pallii, nucleus accumbens, bed nucleus striae terminalis and nucleus interpeduncularis. In the preoptic area/hypothalamus, high levels of binding were clearly present in all areas that contain gonadotropin releasing hormone cells or fibers. In the pons and mesencephalon, moderate levels of binding were associated with catecholaminergic areas such as the area ventralis tegmentalis (area ventralis of Tsai) and the locus coeruleus. Saturation analysis demonstrated the presence of a higher number of binding sites in females than in males in the cerebellar molecular layer, nucleus infundibularis and nucleus anterior medialis. This latter difference was confirmed in the one point assays that also identified higher levels of specific binding in the nucleus suprachiasmaticus pars medialis of males as compared with females. These anatomical data suggest a possible implication of non- N-methyl- d-aspartate receptors in the synthesis and/or release of both gonadotropin releasing hormone and catecholaminergic neurotransmitters that should now be tested by pharmacological experiments.