Abstract:
:Although dopamine (DA) translocation by the DA transporter (DAT) requires Na+, a role for Na+ in the DA recognition step in the translocation cycle has been questioned. Thus, when binding techniques were used to indirectly measure the affinity of DA for DAT via its potency in inhibiting cocaine analog binding, no stimulation of DA binding was observed when assay temperature was at or below room temperature. The present work describes the use of 3H-labeled cocaine analogs for assays at 37 degrees C. When there is sufficient Na+ in the medium (> or =25 mM), [3H]2beta-carbomethoxy-3beta-(4-iodophenyl)tropane ([3H]CIT) is an excellent radioligand to label human DAT with high affinity in membrane preparations of HEK-293 cells expressing the transporter. However, at 0 and 5 mM of Na+, appreciable binding of [3H]CIT occurs to proteins other than DAT, hampering accurate assessment of DAT-associated binding. No such problems occur with the binding of the 4-fluoro homolog of [3H]CIT, [3H]CFT at 37 degrees C, and this radioligand can be used at low [Na+], provided enough protein is present in the assay. The application of these assays show that, in contrast to the strong Na+ dependency of the binding of CFT, the substrates DA, D-amphetamine, p-tyramine, and DL-octopamine are not stimulated by Na+. This demonstrates that lack of Na+ stimulation of binding of substrates, including DA to DAT, in membrane preparations at room temperature is not caused by the reduced fluidity of the frozen state of the hydrocarbon membrane interior at this temperature as compared with the liquid-expanded state at 37 degrees C.
journal_name
J Neurosci Methodsjournal_title
Journal of neuroscience methodsauthors
Wang LC,Cui XN,Chen N,Reith MEdoi
10.1016/s0165-0270(03)00230-9subject
Has Abstractpub_date
2003-12-30 00:00:00pages
27-33issue
1-2eissn
0165-0270issn
1872-678Xpii
S0165027003002309journal_volume
131pub_type
杂志文章abstract:BACKGROUND:Electroencephalogram (EEG) signals are often corrupted with unintended artifacts which need to be removed for extracting meaningful clinical information from them. Typically a priori knowledge of the nature of the artifacts is needed for such purpose. Artifact contamination of EEG is even more prominent for ...
journal_title:Journal of neuroscience methods
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abstract::The physical basis for electrical stimulation of excitable tissue, as used by electrophysiological researchers and clinicians in functional electrical stimulation, is presented with emphasis on the fundamental mechanisms of charge injection at the electrode/tissue interface. Faradaic and non-Faradaic charge transfer m...
journal_title:Journal of neuroscience methods
pub_type: 杂志文章,评审
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abstract::We describe a much simplified high-performance liquid chromatography (HPLC) method for the measurement of caffeine in plasma and brain. A particularly attractive feature of this method is that a simple methanol/water (60:40) mobile phase can be used both for plasma and brain samples. In addition, the method is compati...
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journal_title:Journal of neuroscience methods
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abstract:BACKGROUND:A key point in developmental biology is to understand how gene expression influences the morphological and dynamical patterns that are observed in living beings. NEW METHOD:In this work we propose a methodology capable of addressing this problem that is based on estimating the mutual information and Pearson...
journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
pub_type: 杂志文章
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journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/0165-0270(91)90051-z
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
pub_type: 杂志文章
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journal_title:Journal of neuroscience methods
pub_type: 杂志文章
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journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/j.jneumeth.2014.12.001
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journal_title:Journal of neuroscience methods
pub_type: 杂志文章
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/0165-0270(91)90136-n
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journal_title:Journal of neuroscience methods
pub_type: 杂志文章
doi:10.1016/0165-0270(96)00072-6
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journal_title:Journal of neuroscience methods
pub_type: 杂志文章
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
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journal_title:Journal of neuroscience methods
pub_type: 杂志文章
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更新日期:2013-03-30 00:00:00