Xia Y, Poosch M S, Whitty C J, Kapatos G, Bannon M J
Department of Psychiatry, Wayne State University School of Medicine, Detroit, Michigan.
Neurochem Int. 1993 Mar;22(3):263-70. doi: 10.1016/0197-0186(93)90054-9.
A previously isolated rat cDNA clone encoding the membrane transporter for the neurotransmitter gamma-aminobutyric acid was expressed in transfected COS cells. The resultant transporter protein was characterized kinetically and pharmacologically. The apparent Kt (6.1 microM) and the pharmacological profile of a neuronal-type transporter observed in these mammalian cells were consistent with previous data obtained in Xenopus laevis oocytes. Post-natal levels of gamma-aminobutyric acid transporter mRNA in rat cerebellum, cerebral cortex and striatum (as measured by nuclease protection assay) transiently exceeded levels present in the adult brain. Human gamma-aminobutyric acid transporter mRNA also was measured by nuclease protection assay using as probe a human transporter cDNA homolog obtained by polymerase chain reaction. These studies suggest that quantitation of rat and human gamma-aminobutyric acid transporter mRNAs may provide a useful index of transporter gene expression.
一个先前分离得到的编码神经递质γ-氨基丁酸膜转运体的大鼠cDNA克隆,在转染的COS细胞中得到表达。对产生的转运体蛋白进行了动力学和药理学特性分析。在这些哺乳动物细胞中观察到的神经元型转运体的表观Kt(6.1微摩尔)和药理学特征,与先前在非洲爪蟾卵母细胞中获得的数据一致。通过核酸酶保护试验测定,大鼠小脑、大脑皮层和纹状体中γ-氨基丁酸转运体mRNA的产后水平短暂超过成年大脑中的水平。还使用通过聚合酶链反应获得的人转运体cDNA同源物作为探针,通过核酸酶保护试验测量了人γ-氨基丁酸转运体mRNA。这些研究表明,对大鼠和人γ-氨基丁酸转运体mRNA的定量分析可能为转运体基因表达提供一个有用的指标。