Dopamine transporter: expression in Xenopus oocytes

George R. Uhl, Bruce O'Hara, Shoichi Shimada, Robert Zaczek, Jess DiGiorgianni, Toshikazu Nishimori

Research output: Contribution to journalArticle

Abstract

Xenopus oocytes can express biologically relevant transport activity after injection of mRNAs encoding several carrier molecules. mRNA from PC12 cells, as well as transcripts from a rat ventral midbrain library, can be expressed in these oocytes and allow them to display pharmalogically specific dopamine uptake. mRNA-injected oocytes incubated with tritiated dopamine contain tritiated dopamine and metabolites; lower amounts of radiolabeled dopamine and more radiolabeled metabolites are found in oocytes co-incubated with cocaine or in water-injected oocytes. Tritiated dopamine uptake into mRNA-injected oocytes is time, sodium, and temperature dependent. It is blocked by cocaine and mazindol, but not by haloperidol. It is not found after injection of mRNA from other brain regions. A size-selected rat midbrain library constructed in the plasma vector pCDM8 yields mRNA transcripts whose injection into oocytes causes cocaine-blockable [3H]dopamine uptake. These findings provide an assay for purification of the dopamine transporter cDNA by sib selection techniques.

Original languageEnglish (US)
Pages (from-to)23-29
Number of pages7
JournalMolecular Brain Research
Volume9
Issue number1-2
DOIs
StatePublished - 1991

Keywords

  • Dopamine transporter
  • mRNA
  • PC12 cell
  • Xenopus oocyte

ASJC Scopus subject areas

  • Molecular Biology
  • Cellular and Molecular Neuroscience

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    Uhl, G. R., O'Hara, B., Shimada, S., Zaczek, R., DiGiorgianni, J., & Nishimori, T. (1991). Dopamine transporter: expression in Xenopus oocytes. Molecular Brain Research, 9(1-2), 23-29. https://doi.org/10.1016/0169-328X(91)90126-I