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Dopamine uptake in serotonin transporter knockout mice by chronoamperometry



Dopamine uptake in serotonin transporter knockout mice by chronoamperometry



Society for Neuroscience Abstracts 27(2): 1870



Mice with a targeted disruption of the serotonin transporter (SERT) gene have been produced to study the role of SERT in the modulation of 5-HT neurotransmission and behavior. In vivo microdialysis using zero net flux has revealed a gene dose-dependent increase in extracellular 5-HT levels in SERT KO mice. In addition, our previous high-speed chronoamperometry studies have demonstrated that 5-HT uptake rates are decreased by about half in SERT+/- KO mice vs wildtype mice; while no 5-HT uptake was observed in SERT-/- KO mice. Since our long-term goal is to characterize neuroadaptation resulting from decreases in SERT function, we have also begun to evaluate possible compensatory changes in dopamine uptake in SERT KO mice using chronoamperometry. Crude synaptosomal DA uptake was characterized using Nafion-coated 30mum carbon fiber electrodes. Surprisingly, DA uptake was also decreased in a gene dose-dependent manner in striatum of SERT KO mice (525+-15, 377+-14 and 309+-17 pmole/mg pro/min in wildtype, SERT+/-, and SERT-/- KO mice, respectively; n=6 per genotype). In a preliminary study, preincubation of striatal synaptosomes with 10uM paroxetine to block SERT resulted in reduced DA uptake in both wildtype and SERT+/- KO mice (256+-28 and 255+-10 pmol/mg pr/min) to the level observed in SERT-/- KO mice. These results indicate that SERT is an important modulator of both 5-HT and DA uptake and that apprx40% of the DA uptake observed in striatum is due to promiscuous SERT activity.

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