THE MOLECULAR-CLONING AND EXPRESSION OF A HUMAN SYNAPTIC VESICLE AMINE TRANSPORTER THAT SUPPRESSES MPP(+) TOXICITY

A synaptic vesicle amine transporter cDNA, termed hSVAT, has been isol ated by the reverse transcription and polymerase chain reaction (PCR) technique from human substantia nigra and subsequent screening of a hu man substantia nigra library. The hSVAT sequence obtained is highly ho mologous to the rat SVAT sequence (92% homology) and is essentially id entical to the human sequence identified recently by Surratt and colle agues [33]. This labelled hSVAT cDNA detected a single band (similar t o 5.0 kb) when used as a probe for Northern analysis of human nigral R NA extract. In situ hybridization studies using hSVAT specific antisen se oligonucleotides showed a strong hybridization signal concentrated over the cells of the substantia nigra pars compacta. This cDNA sequen ce when expressed in chinese hamster ovary (CHO) cells conferred resis tance to MPP(+) the toxic metabolite of MPTP and cells containing it a ccumulated dopamine.