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Fluorescent Saxitoxins for Live Cell Imaging of Single Voltage-Gated Sodium Ion Channels beyond the Optical Diffraction Limit

Ondrus, Alison E. and Lee, Hsiao-lu D. and Iwanaga, Shigeki and Parsons, William H. and Andresen, Brian M. and Moerner, W. E. and Du Bois, J. (2012) Fluorescent Saxitoxins for Live Cell Imaging of Single Voltage-Gated Sodium Ion Channels beyond the Optical Diffraction Limit. Chemistry and Biology, 19 (7). pp. 902-912. ISSN 1074-5521. PMCID PMC3731772. doi:10.1016/j.chembiol.2012.05.021.

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A desire to better understand the role of voltage-gated sodium channels (NaVs) in signal conduction and their dysregulation in specific disease states motivates the development of high precision tools for their study. Nature has evolved a collection of small molecule agents, including the shellfish poison (+)-saxitoxin, that bind to the extracellular pore of select NaV isoforms. As described in this report, de novo chemical synthesis has enabled the preparation of fluorescently labeled derivatives of (+)-saxitoxin, STX-Cy5, and STX-DCDHF, which display reversible binding to NaVs in live cells. Electrophysiology and confocal fluorescence microscopy studies confirm that these STX-based dyes function as potent and selective NaV labels. The utility of these probes is underscored in single-molecule and super-resolution imaging experiments, which reveal NaV distributions well beyond the optical diffraction limit in subcellular features such as neuritic spines and filopodia.

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URLURL TypeDescription Information Information - Video CentralArticle
Ondrus, Alison E.0000-0002-6023-3290
Du Bois, J.0000-0001-7847-1548
Additional Information:Ⓒ 2012 Elsevier Ltd. Received: February 24, 2012. Revised: May 24, 2012. Accepted: May 25, 2012. Published: July 26, 2012. A.E.O., H.L., S.I., B.M.A., W.E.M., and J.D. designed research; A.E.O., H.L., S.I., and W.H.P. performed research; A.E.O., H.L., and S.I. analyzed data; and A.E.O., H.L., W.E.M., and J.D. wrote the paper. The authors thank Professor R.J. Twieg and J.C. Williams for the gift of DCDHF-NHS. We are grateful to Professor Merritt Maduke for allowing use of her electrophysiology equipment and for many helpful discussions. This work was supported in part by R01-GM086196 (to W.E.M.) and R01-NS45684 (to J.D.) from the National Institute of General Medical Sciences and the National Institute of Neurological Disorders and Stroke, respectively, and by a grant from the Tobacco-Related Disease Research Program (to J.D.). W.H.P. is the recipient of a Stanford Interdisciplinary Graduate Fellowship. J.D. is a cofounder of SiteOne Therapeutics, Inc.
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California Tobacco-Related Disease Research ProgramUNSPECIFIED
Stanford UniversityUNSPECIFIED
Issue or Number:7
PubMed Central ID:PMC3731772
Record Number:CaltechAUTHORS:20170901-132021426
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ID Code:81079
Deposited By: Donna Wrublewski
Deposited On:05 Sep 2017 19:21
Last Modified:25 Apr 2022 20:49

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