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dc.contributor.authorHardy, Matthew E.
dc.contributor.authorPollard, C.E.
dc.contributor.authorSmall, B.G.
dc.contributor.authorBridgland-Taylor, M.
dc.contributor.authorWoods, A.J.
dc.contributor.authorValentin, J.-P.
dc.contributor.authorAbi-Gerges, N.
dc.date.accessioned2018-07-25T11:21:43Z
dc.date.available2018-07-25T11:21:43Z
dc.date.issued2009
dc.identifier.citationHardy ME, Pollard CE, Small BG, Bridgland-Taylor M, Harmer AR, Woods AJ, Valentin J-P and Abi-Gerges N (2009) Validation of a voltage-sensitive dye (di-4-ANEPPS)-based method for assessing drug-induced delayed repolarisation in Beagle dog left ventricular midmyocardial myocytes. Journal of Pharmacological and Toxicological Methods. Focused Issue on Methods in Safety Pharmacology. 60: 94-106.en_US
dc.identifier.urihttp://hdl.handle.net/10454/16509
dc.descriptionnoen_US
dc.description.abstractEvaluation of drug candidates in in-vitro assays of action potential duration (APD) is one component of preclinical safety assessment. Current assays are limited by technically-demanding, time-consuming electrophysiological methods. This study aimed to assess whether a voltage-sensitive dye-based assay could be used instead. Methods Optical APs were recorded using di-4-ANEPPS in electrically field stimulated Beagle left ventricular midmyocardial myocytes (LVMMs). Pharmacological properties of di-4-ANEPPS on the main cardiac ion channels that shape the ventricular AP were investigated using IonWorks™ and conventional electrophysiology. Effects of 9 reference drugs (dofetilide, E4031, d-sotalol, ATXII, cisapride, terfenadine, alfuzosin, diltiazem and pinacidil) with known APD-modulating effects were assessed on optically measured APD at 1 Hz. Results Under optimum conditions, 0.1 μM di-4-ANEPPS could be used to monitor APs paced at 1 Hz during nine, 5 s exposures without altering APD. di-4-ANEPPS had no effect on either hIERG, hINa, hIKs and hIto currents in transfected CHO cells (up to 10 µM) or ICa,L current in LVMMs (at 16 µM). di-4-ANEPPS had no effect on APs recorded with microelectrodes at 1 or 0.5 Hz over a period of 30 min di-4-ANEPPS displayed the sensitivity to record changes in optically measured APD in response to altered pacing frequencies and sequential vehicle additions did not affect the optically measured APD. APD data obtained with 9 reference drugs were as expected except (i) d-sotalol-induced increases in duration were smaller than those caused by other IKr blockers and (ii) increases in APD were not detected using low concentrations of terfenadine. Discussion Early in drug discovery, the di-4-ANEPPS-based method can reliably be used to assess drug effects on APD as part of a cardiac risk assessment strategy.en_US
dc.language.isoenen_US
dc.relation.isreferencedbyhttps://doi.org/10.1016/j.vascn.2009.03.005en_US
dc.subjectAction potential durationen_US
dc.subjectCardiac ion channelsen_US
dc.subjectDog speciesen_US
dc.subjectIonWorks (TM)en_US
dc.subjectLeft ventricular midmyocardial myocytesen_US
dc.subjectMethodsen_US
dc.subjectVoltage-sensitive dyeen_US
dc.titleValidation of a voltage-sensitive dye (di-4-ANEPPS)-based method for assessing drug-induced delayed repolarisation in Beagle dog left ventricular midmyocardial myocytesen_US
dc.status.refereedyesen_US
dc.typeArticleen_US
dc.type.versionNo full-text in the repositoryen_US


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