[3H] Ethynylbicycloorthobenzoate ([3H] EBOB) Binding in Native and Recombinant GABAA Receptors

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dc.contributor.advisorDillon, Glenn
dc.contributor.committeeMemberMartin, Michael
dc.contributor.committeeMemberde Fiebre, Christopher
dc.creatorYagle, Monica A.
dc.date.accessioned2019-08-22T21:42:11Z
dc.date.available2019-08-22T21:42:11Z
dc.date.issued2000-05-01
dc.date.submitted2014-04-14T08:48:04-07:00
dc.description.abstractYagle, Monica A., [3H] Ethynylbicycloorthobenzoate ([3H] EBOB) Binding in Native and Recombinant GABAA Receptors. Master of Science (Pharmacology), May 2000, 59 pp., 3 tables, 7 illustrations, bibliography, 75 titles. Modulation of the GABAA receptor has been studied with noncompetitive convulsant ligands such as tert-butylbicyclophosphorothionate (TBPS) and picrotoxin (PTX). EBOB is a more recently developed ligand that appears to bind in the same region of the channel at TBPS, but with a higher affinity. While only a few studies have examined the binding of EBOB to vertebrate brain tissue and insect preparations, none have examined potential subunit-dependent binding of EBOB. We have thus examined [3H] EBOB binding in rat cerebellum and HEK293 cells stably expressing human α1β2γ2, human α2β2γ2, and rat α6β2γ2 GABAA receptors. For comparison, [35S] TBPS binding was also examined in α1β2γ2 receptors. Saturation and Scatchard analyses revealed saturable [3H] EBOB binding at one site in all tissue preparations with Kd values ranging from 3 to 9nM. [3H] EBOB binding, like [35S] TBPS binding was inhibited by the CNS convulsants dieldrin, lindane, tert-butylbicyclophosphorothionate (TBOB), PTX, TBPS, and pentylenetetrazole (PTZ) at one site in a concentration dependent fashion. Affinities were in the high nM to low μM range for all compounds except PTZ (low mM range). GABA modulated [3H] EBOB binding in a biphasic manner in α1β2γ2 receptors with a 100-fold difference between stimulatory and inhibitory affinities. Inhibition of GABA-mediated current by TBOB in α1β2γ2 receptors resulted in a functional IC50 of 0.2 μM, in agreement with binding study results. Differences seen in binding between the different receptor subtypes examined suggest that some characteristics of EBOB binding are subunit dependent. In addition, we have shown that [3H] EBOB is a useful ligand in the study of recombinant GABAA receptors and that results obtained with [3H] EBOB are comparable to those obtained with [35S] TBPS.
dc.format.mimetypeapplication/pdf
dc.identifier.urihttps://hdl.handle.net/20.500.12503/29523
dc.language.isoen
dc.provenance.legacyDownloads0
dc.subjectCell and Developmental Biology
dc.subjectCell Biology
dc.subjectCells
dc.subjectCellular and Molecular Physiology
dc.subjectChemical and Pharmacologic Phenomena
dc.subjectChemicals and Drugs
dc.subjectLife Sciences
dc.subjectMedical Cell Biology
dc.subjectMedical Neurobiology
dc.subjectMedical Pharmacology
dc.subjectMedical Sciences
dc.subjectMedicine and Health Sciences
dc.subjectMolecular and Cellular Neuroscience
dc.subjectNeuroscience and Neurobiology
dc.subjectOther Cell and Developmental Biology
dc.subjectOther Neuroscience and Neurobiology
dc.subjectPharmacology
dc.subjectEthynylbicycloorthobenzoate
dc.subject[3H] EBOB
dc.subjectnative GABAA receptors
dc.subjectrecombinant GABAA receptors
dc.subjectnoncompetitive convulsant ligands
dc.subjecttert-butylbicyclophosphorothionate
dc.subjectTBPS
dc.subjectpicrotoxin
dc.subjectPTX
dc.subjectsub-unit dependent binding
dc.subjectrat cerebellum
dc.subjectHEK293 cells
dc.title[3H] Ethynylbicycloorthobenzoate ([3H] EBOB) Binding in Native and Recombinant GABAA Receptors
dc.typeThesis
dc.type.materialtext
thesis.degree.departmentGraduate School of Biomedical Sciences
thesis.degree.disciplinePharmacology and Neuroscience
thesis.degree.grantorUniversity of North Texas Health Science Center at Fort Worth
thesis.degree.nameMaster of Science

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