RESUMO
RNA isolated from both rat phaeochromocytoma (PC12) cells and bovine adrenal medulla was size-selected and injected into oocytes of Xenopus laevis. Expression of the neuronal noradrenaline transporter was assayed 3 days after injection by measuring the nisoxetine-sensitive noradrenaline uptake. A 2 kilobase fraction of either total RNA derived from both tissues or of polyadenylated mRNA obtained from bovine adrenal medulla caused Na(+)-dependent transport of 3H-noradrenaline in the injected oocytes.
Assuntos
Proteínas de Transporte/metabolismo , Simportadores , Xenopus laevis/metabolismo , Neoplasias das Glândulas Suprarrenais/metabolismo , Animais , Proteínas de Transporte/genética , Células Cultivadas , Feminino , Regulação da Expressão Gênica , Norepinefrina/metabolismo , Proteínas da Membrana Plasmática de Transporte de Norepinefrina , Oócitos/metabolismo , Feocromocitoma/metabolismo , RNA/isolamento & purificação , RNA/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Neoplásico/metabolismo , Ratos , Trítio , Células Tumorais CultivadasRESUMO
The neuronal noradrenaline transporter was partially purified by means of low and high pressure liquid chromatography using anion exchange, gel filtration and lectin affinity columns. A protein characterized by a molecular weight of 50-53 kilodalton was enriched; it may represent the transporter or a component of it. In addition, a RNA fraction characterized by a mean size of 2 kilobases was isolated from PC12 rat phaeochromocytoma cells and from bovine adrenal medulla; this RNA fraction caused expression of the noradrenaline transporter after microinjection into Xenopus laevis oocytes.