Monitoring of compound resting membrane potentials of cell cultures with ratiometric genetically encoded voltage indicators

GND
1256615838
Affiliation
Center for Molecular Biomedicine, Department of Biophysics, Friedrich Schiller University Jena and Jena University Hospital, Jena, Germany
Rühl, Philipp;
GND
1233910345
Affiliation
Center for Molecular Biomedicine, Department of Biophysics, Friedrich Schiller University Jena and Jena University Hospital, Jena, Germany
Langner, Johanna M.;
GND
1316985644
Affiliation
Center for Molecular Biomedicine, Department of Biophysics, Friedrich Schiller University Jena and Jena University Hospital, Jena, Germany
Reidel, Jasmin;
GND
1145802958
Affiliation
Center for Molecular Biomedicine, Department of Biophysics, Friedrich Schiller University Jena and Jena University Hospital, Jena, Germany
Schönherr, Roland;
Affiliation
Department of Physiology, University of Pennsylvania, Philadelphia, USA
Hoshi, Toshinori;
GND
1203132905
ORCID
0000-0002-4144-0251
Affiliation
Center for Molecular Biomedicine, Department of Biophysics, Friedrich Schiller University Jena and Jena University Hospital, Jena, Germany
Heinemann, Stefan H.

Abstract The cellular resting membrane potential ( V m ) not only determines electrical responsiveness of excitable cells but also plays pivotal roles in non-excitable cells, mediating membrane transport, cell-cycle progression, and tumorigenesis. Studying these processes requires estimation of V m , ideally over long periods of time. Here, we introduce two ratiometric genetically encoded V m indicators, rArc and rASAP, and imaging and analysis procedures for measuring differences in average resting V m between cell groups. We investigated the influence of ectopic expression of K + channels and their disease-causing mutations involved in Andersen-Tawil (Kir2.1) and Temple-Baraitser (K V 10.1) syndrome on median resting V m of HEK293T cells. Real-time long-term monitoring of V m changes allowed to estimate a 40–50 min latency from induction of transcription to functional Kir2.1 channels in HEK293T cells. The presented methodology is readily implemented with standard fluorescence microscopes and offers deeper insights into the role of the resting V m in health and disease.

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