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U‐373 MG glioblastoma and IMR‐32 neuroblastoma cell lines express the dopamine and vesicular monoamine transporters
Author(s) -
Koutsilieri E.,
Kornhuber J.,
Degen H.J.,
Lesch K.P.,
Sopper S.,
Meulen V. ter,
Riederer P.
Publication year - 1996
Publication title -
journal of neuroscience research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.72
H-Index - 160
eISSN - 1097-4547
pISSN - 0360-4012
DOI - 10.1002/(sici)1097-4547(19960801)45:3<269::aid-jnr8>3.0.co;2-b
Subject(s) - monoamine neurotransmitter , dopamine , vesicular monoamine transporter 2 , glioblastoma , neuroblastoma , chemistry , dopamine transporter , pharmacology , biology , cell culture , cancer research , neuroscience , biochemistry , dopaminergic , genetics , serotonin , receptor
The U‐373 MG glioblastoma and the IMR‐32 neuroblastoma cell lines were found to express the dopamine (DA) and vesicular monoamine transporters, using reverse transcriptase‐polymerase chain reaction (RT‐PCR). To further characterize the DA transporter, [ 3 H]GBR‐12935 binding and [ 3 H]DA uptake studies were performed. Specific binding of [ 3 H]GBR‐12935 to U‐373 MG and IMR‐32 cells is saturable as saturation experiments indicated. Scatchard analysis revealed two binding sites on U‐373 MG as well as on IMR‐32 cells. The high‐affinity sites exhibited a K D of 2.95 and 0.42 nM and a B max of 6.4 and 0.83 fmol/mg protein for U‐373 MG and IMR‐32 cells, respectively. The low‐affinity sites exhibited a K D of 144 and 251 nM and a B max of 37.5 and 119 fmol/mg protein for the same cells, respectively. The high‐affinity binding of both types of cells probably represents the “classic” DA uptake site identified in other studies from human and rat striatal membranes or synaptosomes, while the low‐affinity binding may represent a mazindol‐insensitive binding site (the “piperazine acceptor site”). [ 3 H]DA uptake was 0.55 ± 0.16 and 1.08 ± 0.33 pmol/mg protein for U‐373 MG and IMR‐32 cells, respectively. Since the DA transporter has been implicated as an important site for drugs and toxins, the above‐mentioned cell lines may be a useful tool in the study of the mechanism of action of DA transporter modulating substances. © 1996 Wiley‐Liss, Inc.