Screening Fluorescent Voltage Indicators with Spontaneously Spiking HEK Cells

Development of improved fluorescent voltage indicators is a key challenge in neuroscience, but progress has been hampered by the low throughput of patch-clamp characterization. We introduce a line of non-fluorescent HEK cells that stably express Na(V) 1.3 and K(IR) 2.1 and generate spontaneous elect...

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Detalles Bibliográficos
Autores principales: Park, Jeehae, Werley, Christopher A., Venkatachalam, Veena, Kralj, Joel M., Dib-Hajj, Sulayman D., Waxman, Stephen G., Cohen, Adam E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
Materias:
Research Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3877367/
https://www.ncbi.nlm.nih.gov/pubmed/24391999
http://dx.doi.org/10.1371/journal.pone.0085221
Descripción
Sumario:Development of improved fluorescent voltage indicators is a key challenge in neuroscience, but progress has been hampered by the low throughput of patch-clamp characterization. We introduce a line of non-fluorescent HEK cells that stably express Na(V) 1.3 and K(IR) 2.1 and generate spontaneous electrical action potentials. These cells enable rapid, electrode-free screening of speed and sensitivity of voltage sensitive dyes or fluorescent proteins on a standard fluorescence microscope. We screened a small library of mutants of archaerhodopsin 3 (Arch) in spiking HEK cells and identified two mutants with greater voltage-sensitivity than found in previously published Arch voltage indicators.

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