Binding of [ 3 H]mazindol to cardiac norepinephrine transporters: kinetic and equilibrium studies
dc.contributor.author | Raffel, David M. | en_US |
dc.contributor.author | Chen, Wei | en_US |
dc.date.accessioned | 2006-09-11T17:37:41Z | |
dc.date.available | 2006-09-11T17:37:41Z | |
dc.date.issued | 2004-07 | en_US |
dc.identifier.citation | Raffel, David M.; Chen, Wei; (2004). "Binding of [ 3 H]mazindol to cardiac norepinephrine transporters: kinetic and equilibrium studies." Naunyn-Schmiedeberg's Archives of Pharmacology 370(1): 9-16. <http://hdl.handle.net/2027.42/46313> | en_US |
dc.identifier.issn | 0028-1298 | en_US |
dc.identifier.issn | 1432-1912 | en_US |
dc.identifier.uri | https://hdl.handle.net/2027.42/46313 | |
dc.identifier.uri | http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=15300361&dopt=citation | en_US |
dc.description.abstract | The norepinephrine transporter (NET) is the carrier that drives the neuronal norepinephrine uptake mechanism (uptake 1 ) in mammalian hearts. The radioligand [ 3 H]mazindol binds with high affinity to NET. In this study, the kinetics of [ 3 H]mazindol binding to NET were measured using a rat heart membrane preparation. Results from these studies were used to set up saturation binding assays designed to measure cardiac NET densities ( B max ) and competitive inhibition assays designed to measure inhibitor binding affinities ( K I ) for NET. Saturation binding assays measured NET densities in rat, rabbit, and canine hearts. Assay reproducibility was assessed and the effect of NaCl concentration on [ 3 H]mazindol binding to NET was studied using membranes from rat and canine hearts. Specificity of [ 3 H]mazindol binding to NET was determined in experiments in which the neurotoxin 6-hydroxydopamine (6-OHDA) was used to selectively destroy cardiac sympathetic nerve terminals in rats. Competitive inhibition studies measured K I values for several NET inhibitors and substrates. In kinetic studies using rat heart membranes, [ 3 H]mazindol exhibited a dissociation rate constant k off =0.0123±0.0007 min −1 and an association rate constant k on =0.0249±0.0019 nM −1 min −1 . In saturation binding assays, [ 3 H]mazindol binding was monophasic and saturable in all cases. Increasing the concentration of NaCl in the assay buffer increased binding affinity significantly, while only modestly increasing B max . Injections of 6-OHDA in rats decreased measured cardiac NET B max values in a dose-dependent manner, verifying that [ 3 H]mazindol binds specifically to NET from sympathetic nerve terminals. Competitive inhibition studies provided NET inhibitor and substrate K I values consistent with previously reported values. These studies demonstrate the high selectivity of [ 3 H]mazindol binding for the norepinephrine transporter in membrane preparations from mammalian hearts. | en_US |
dc.format.extent | 162953 bytes | |
dc.format.extent | 3115 bytes | |
dc.format.mimetype | application/pdf | |
dc.format.mimetype | text/plain | |
dc.language.iso | en_US | |
dc.publisher | Springer-Verlag | en_US |
dc.subject.other | Norepinephrine Transporter | en_US |
dc.subject.other | LifeSciences | en_US |
dc.subject.other | Meta -Iodobenzylguanidine | en_US |
dc.subject.other | Sympathetic Nervous System | en_US |
dc.subject.other | Positron Emission Tomography | en_US |
dc.subject.other | Meta -Hydroxyephedrine | en_US |
dc.title | Binding of [ 3 H]mazindol to cardiac norepinephrine transporters: kinetic and equilibrium studies | en_US |
dc.type | Article | en_US |
dc.subject.hlbsecondlevel | Pharmacy and Pharmacology | en_US |
dc.subject.hlbtoplevel | Health Sciences | en_US |
dc.description.peerreviewed | Peer Reviewed | en_US |
dc.contributor.affiliationum | Division of Nuclear Medicine, Department of Radiology, University of Michigan Medical School, 3480 Kresge III Building, Ann Arbor, MI, 48109-0552, USA | en_US |
dc.contributor.affiliationum | Division of Nuclear Medicine, Department of Radiology, University of Michigan Medical School, 3480 Kresge III Building, Ann Arbor, MI, 48109-0552, USA | en_US |
dc.contributor.affiliationumcampus | Ann Arbor | en_US |
dc.identifier.pmid | 15300361 | en_US |
dc.description.bitstreamurl | http://deepblue.lib.umich.edu/bitstream/2027.42/46313/1/210_2004_Article_949.pdf | en_US |
dc.identifier.doi | http://dx.doi.org/10.1007/s00210-004-0949-y | en_US |
dc.identifier.source | Naunyn-Schmiedeberg's Archives of Pharmacology | en_US |
dc.owningcollname | Interdisciplinary and Peer-Reviewed |
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