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 isolated by the reverse transcription and polymerase chain reaction (PCR) technique from human substantia nigra and subsequent screening of a human substantia nigra library. The hSVAT sequence obtained is highly homologous to the rat SVAT sequence (92% homology) and is essentially identical to the human sequence identified recently by Surratt and colleagues [33]. This labelled hSVAT cDNA detected a single band (approximately 5.0 kb) when used as a probe for Northern analysis of human nigral RNA extract. In situ hybridization studies using hSVAT specific antisense oligonucleotides showed a strong hybridization signal concentrated over the cells of the substantia nigra pars compacta. This cDNA sequence when expressed in chinese hamster ovary (CHO) cells conferred resistance to MPP+ the toxic metabolite of MPTP and cells containing it accumulated dopamine.
Liu
L
L
MRC Molecular Neuroscience Group, Department of Neurobiology, Babraham Institute, Cambridge, UK.
Xu
W
W
Harrington
K A
KA
Emson
P C
PC
D015655:1-Methyl-4-phenylpyridinium / Q000037:antagonists & inhibitors / Q000633:toxicity*; D000588:Amines / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001692:Biological Transport / Q000502:physiology; D016466:CHO Cells; D002352:Carrier Proteins / Q000096:biosynthesis*; D003001:Cloning, Molecular; D006224:Cricetinae; D018076:DNA, Complementary / Q000302:isolation & purification; D015698:Genomic Library; D006801:Humans; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000096:biosynthesis*; D013378:Substantia Nigra / Q000737:chemistry; D013572:Synaptic Vesicles; D014162:Transfection
D000588:Amines; D002352:Carrier Proteins; D018076:DNA, Complementary; D009419:Nerve Tissue Proteins; D015655:1-Methyl-4-phenylpyridinium