Abstract
Fluorescence-based biosensors for membrane voltage allow dynamic optical recording of neuronal activity. Interestingly, the development of genetically encoded voltage indicators constitutes a good alternative to classical voltage-sensitive dyes, since they allow overcoming some of the inherent problems (e.g., optical noise, etc.) associated to organic compounds. Here, we show the use of a genetically encoded voltage-sensitive fluorescent protein (VSFP), namely the VSFP2.32. This biosensor contains a mCerulean and Citrine tandem, which can engage in a constitutive fluorescent resonance energy transfer (FRET) process. We first expressed VSFP2.32 in hippocampal cultured neurons. And, subsequently, we monitored membrane voltage alterations in single neurons by recording (in a real-time mode) VSFP2.32 conformation-mediated FRET changes.
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Acknowledgments
This work was supported by Ministerio de Ciencia, Innovación y Universidades–Agencia Estatal de Investigación-FEDER-UE (SAF2017-87349-R MICIU/AEI/FEDER/UE) and Generalitat de Catalunya (2017SGR1604).We thank Centres de Recerca de Catalunya (CERCA) Programme/Generalitat de Catalunya for IDIBELL institutional support. We thank Esther Castaño and Benjamín Torrejón from the CCiT-Bellvitge Campus of the University of Barcelona.
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Fernández-Dueñas, V., Morató, X., Knöpfel, T., Ciruela, F. (2021). Dynamic Recording of Membrane Potential from Hippocampal Neurons by Using a Fluorescence Resonance Energy Transfer-Based Voltage Biosensor. In: Lujan, R., Ciruela, F. (eds) Receptor and Ion Channel Detection in the Brain. Neuromethods, vol 169. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1522-5_31
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DOI: https://doi.org/10.1007/978-1-0716-1522-5_31
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