RAT BRAIN SYNAPTIC VESICLES: UPTAKE SPECIFICITIES OF [³H]NOREPINEPHRINE AND [³H]SEROTONIN IN PREPARATIONS FROM WHOLE BRAIN AND BRAIN REGIONS¹

Abstract

Abstract A fraction containing neurotransmitter storage vesicles was isolated from rat whole brain and brain regions, and the uptakes of [ 3 H]norepinephrine and [ 3 H]serotonin were determined in vitro. Norepinephrine uptake in vesicle preparations from corpus striatum was higher than in prep arations from cerebral cortex, and uptake in vesicles from the remainder (midbrain + brainstem + cerebellum) was intermediate. The K m for norepinephrine uptake was the same in the three brain regions, but the regions differed in maximal uptake capacity by factors which paralleled total catecholamine concentration rather than content of norepinephrine alone. Intracisternal administration of 6‐hydroxydopamine, but not of 5,6‐dihydroxytryptamine, reduced vesicular norepinephrine uptake, and pretreat‐ment with desmethylimipramine (which protects specifically norepinephrine neurons but not dopamine neurons from the 6‐hydroxydopamine) only partially prevented the loss of vesicular norepinephrine uptake. These studies indicate that uptake of norepinephrine by rat brain vesicle preparations occurs in vesicles from norepinephrine and dopamine neurons, but probably not in vesicles from serotonin neurons. Uptake of serotonin by brain vesicle preparations exhibited time, temperature and ATP‐Mg 2+ requirements nearly identical to those of norepinephrine uptake. The affinity of serotonin uptake matched that of serotonin for inhibition of norepinephrine uptake, and the maximal capacity was the same for serotonin as for norepinephrine. Norepinephrine, dopamine and reserpine inhibited serotonin uptake in a purely competitive fashion, with K i s similar to those for inhibition of norepinephrine uptake. Whereas 5,6‐dihydroxytryptamine treatment reduced synaptosomal serotonin uptake but not vesicular serotonin uptake, 6‐hydroxydopamine reduced vesicular serotonin uptake in the absence of reductions in synaptosomal serotonin uptake. Thus, in this preparation, serotonin appears to be taken up in vitro into catecholamine vesicles, rather than into serotonin vesicles.