Uptake and release of neurotransmitters.
The availability of clonal cell lines for norepinephrine, dopamine, and serotonin transporters allows the characterization of drug interactions with transporter recognition sites using radioligands, as well as the characterization of drug effects on selective transporter-mediated uptake and release of substrate. In addition to clonal cell lines, synaptosomes prepared from specific brain regions can be used to conduct these studies without interference by endogenous transporters or binding proteins that are present in other tissues. This unit presents protocols for uptake and release of tritiated substrates using intact cells (either detached or in suspension) or synaptosomes. An HPLC procedure for electrochemical detection of nonradiolabeled substrates is also provided. Time-dependent release can also be measured in assays involving real-time sampling.
Janowsky
A
A
Oregon Health Sciences University and Veterans Affairs Medical Center, Portland, Oregon, USA.
Neve
K
K
Eshleman
A J
AJ
D000818:Animals; D002460:Cell Line / Q000187:drug effects / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002999:Clone Cells / Q000187:drug effects / Q000378:metabolism; D003342:Corpus Striatum / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D004298:Dopamine / Q000378:metabolism; D050483:Dopamine Plasma Membrane Transport Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004563:Electrochemistry; D006801:Humans; D007668:Kidney; D018377:Neurotransmitter Agents / Q000378:metabolism*; D011619:Psychotropic Drugs / Q000494:pharmacology; D011869:Radioligand Assay; D051381:Rats; D011954:Receptors, Dopamine / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D013574:Synaptosomes / Q000187:drug effects / Q000378:metabolism
D050483:Dopamine Plasma Membrane Transport Proteins; D018377:Neurotransmitter Agents; D011619:Psychotropic Drugs; D011954:Receptors, Dopamine; D011993:Recombinant Fusion Proteins; D004298:Dopamine