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    Can optical recordings of membrane potential be used to screen for drug-induced action potential prolongation in single cardiac myocytes?

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    Publication date
    2006
    Author
    Hardy, Matthew E.
    Lawrence, C.L.
    Standen, N.B.
    Rodrigo, G.C.
    Keyword
    Action potential
    Arrythmia
    Cardiac muscle
    Guinea-pig myocyte
    Voltage-sensitive dyes
    Peer-Reviewed
    yes
    
    Metadata
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    Abstract
    Introduction: Potential-sensitive dyes have primarily been used to optically record action potentials (APs) in whole heart tissue. Using these dyes to record drug-induced changes in AP morphology of isolated cardiac myocytes could provide an opportunity to develop medium throughout assays for the pharmaceutical industry. Ideally, this requires that the dye has a consistent and rapid response to membrane potential, is insensitive to movement, and does not itself affect AP morphology. Materials and methods: We recorded the AP from isolated adult guinea-pig ventricular myocytes optically using di-8-ANEPPS in a single-excitation dual-emission ratiometric system, either separately in electrically field stimulated myocytes, or simultaneously with an electrical AP recorded with a patch electrode in the whole-cell bridge mode. The ratio of di-8-ANEPPS fluorescence signal was calibrated against membrane potential using a switch-clamp to voltage clamp the myocyte. Results: Our data show that the ratio of the optical signals emitted at 560/620 nm is linearly related to voltage over the voltage range of an AP, producing a change in ratio of 7.5% per 100mV, is unaffected by cell movement and is identical to the AP recorded simultaneously with a patch electrode. However, the APD90 recorded optically in myocytes loaded with di-8-ANEPPS was significantly longer than in unloaded myocytes recorded with a patch electrode (355.6 ± 13.5 vs. 296.2 ± 16.2ms; p< 0.01). Despite this effect, the apparent IC50 for cisapride, which prolongs the AP by blocking IKr, was not significantly different whether determined optically or with a patch electrode (91 ± 46 vs. 81 ± 20 nM). Discussion: These data show that the optical AP recorded ratiometrically using di-8- ANEPPS from a single ventricular myocyte accurately follows the action potential morphology. This technique can be used to estimate the AP prolonging effects of a compound, although di-8-ANEPPS itself prolongs APD90. Optical dyes require less technical skills and are less invasive than conventional electrophysiological techniques and, when coupled to ventricular myocytes, decreases animal usage and facilitates higher throughput assays.
    URI
    http://hdl.handle.net/10454/16510
    Version
    No full-text in the repository
    Citation
    Hardy ME, Lawrence CL, Standen NB and Rodrigo GC (2006) Can optical recordings of membrane potential be used to screen for drug-induced action potential prolongation in single cardiac myocytes? Journal of Pharmacolgical and Toxicolical Methods. 54(2): 173-82.
    Link to publisher’s version
    https://doi.org/10.1016/j.vascn.2006.02.013
    Type
    Article
    Collections
    Life Sciences Publications

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