[H]5-HT binding sites and 5-HT-sensitive adenylate cyclase in glial cell membrane fraction

Glial cell membrane fractions were prepared using glial cells preparations isolated from horse brain striatum. [³H]5-HT binding was measured by the filtration technique and the adenylate cyclase activity determined by measuring the cAMP production using a radioimmunoassay.Serotonin binds to glial membrane fractions with an affinity corresponding to a dissociation constantK_d = 10 nM. The corresponding site is serotoninergic specific: [³H]5-HT binding is inhibited by 5-HT agonists (5 OH NM-DMT, 5-MeOHT, 5-MeOH-DMT, NN-DMT) or antagonists (cinanserine, cyproheptadine, methysergide, LSD) and not (or poorly) inhibited by non-serotoninergic related drugs. The population of sites binding 5-HT, present in neuronal membrane preparations and determined in parallel assays is distinct from that observed in glial preparations. The glial membrane fractions contains an adenylate cyclase activated by 5-HT with an apparent affinity constant close to 1 μM. It is serotonin-specific and clearly distinct from the DA-stimulated adenylate cyclase present in the same preparation.The sites binding 5-HT and activating the adenylate cyclase with low affinities might be directly related. This system, clearly distinct from the postsynaptosomal serotoninergic receptor, represents presumably a glial serotoninergic receptor; however, it cannot be totally excluded that these sites may refer to presynaptic membranes.