TY - JOUR
T1 - Dopamine transporter
T2 - expression in Xenopus oocytes
AU - Uhl, George R.
AU - O'Hara, Bruce
AU - Shimada, Shoichi
AU - Zaczek, Robert
AU - DiGiorgianni, Jess
AU - Nishimori, Toshikazu
PY - 1991
Y1 - 1991
N2 - 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.
AB - 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.
KW - Dopamine transporter
KW - mRNA
KW - PC12 cell
KW - Xenopus oocyte
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U2 - 10.1016/0169-328X(91)90126-I
DO - 10.1016/0169-328X(91)90126-I
M3 - Article
C2 - 1850075
AN - SCOPUS:0025917589
SN - 0169-328X
VL - 9
SP - 23
EP - 29
JO - Molecular Brain Research
JF - Molecular Brain Research
IS - 1-2
ER -