Evidence for the presynaptic localization of opiate binding sites on striatal efferent fibers

Abstract

Using quantitative receptor autoradiography, [ 3H] d-Ala- d-Leu-enkephalin (DADL) and [ 3H]naloxone binding were studied in rat striatum and striatal projection areas (globus pallidus (GP) and substantia nigra pars reticulata (SN r)) after unilateral striatal kainic acid lesions. [ 3H]DADL and [ 3H]naloxone binding were each examined by two methods. Initially, [ 3H]DADL binding was performed in 50 mM Tris-HCl (pH 7.4), 30 mM NaCl, 3 mM manganese acetate and 2 μM GTP; [ 3H]naloxone binding was carried out in 50 mM Tris-HCl (pH 7.4) and 100 mM NaCl. Subsequent studies were carried out in 150 mM Tris-HCl (pH 7.4) and either [ 3H]DADL plus 500 nM morphiceptin (to block [ 3H]DADL binding to mu receptors) or [ 3H]naloxone plus 10 nM delta receptor peptide (to block [ 3H]naloxone binding to delta receptors). At one and eight weeks in the lesioned striatum, [ 3H]DADL binding was reduced by 70% and 82%, respectively, when compared to the control side. [ 3H]Naloxone binding was reduced by 35% and 20%. In GP and SN r, [ 3H]DADL binding was reduced by 31% and 41%, respectively, at one week and 27% and 26% at eight weekds. [ 3H]Nalaxone binding was reduced 19% in GP at eight weeks. A parsimonious explanation of these results is that opiate binding sites are located on presynaptic terminals of striatal efferent fibers to globus pallidus and substantia nigra pars reticulata as well as on local striatal axon collaterals. Since opiate peptides have recently been found to coexist with GABA in some striatal neurons, opiate peptides may play a role in striatal function by controlling GABA release from striatal efferent fibers. It is possible that pallidal and nigral opiate binding could be utilized as a marker for striatal terminals.