Article abstract
Nature Methods 5, 797 - 804 (2008)
Published online: 10 August 2008 | doi:10.1038/nmeth.1242
Single-spike detection in vitro and in vivo with a genetic Ca2+ sensor
Damian J Wallace1,6, Stephan Meyer zum Alten Borgloh2,6, Simone Astori2,6, Ying Yang2, Melanie Bausen2, Sebastian Kügler3, Amy E Palmer4, Roger Y Tsien4, Rolf Sprengel2, Jason N D Kerr1, Winfried Denk2 & Mazahir T Hasan2,5
Abstract
Measurement of population activity with single-action-potential, single-neuron resolution is pivotal for understanding information representation and processing in the brain and how the brain's responses are altered by experience. Genetically encoded indicators of neuronal activity allow long-term, cell type–specific expression. Fluorescent Ca2+ indicator proteins (FCIPs), a main class of reporters of neural activity, initially suffered, in particular, from an inability to report single action potentials in vivo. Although suboptimal Ca2+-binding dynamics and Ca2+-induced fluorescence changes in FCIPs are important factors, low levels of expression also seem to play a role. Here we report that delivering D3cpv, an improved fluorescent resonance energy transfer–based FCIP, using a recombinant adeno-associated virus results in expression sufficient to detect the Ca2+ transients that accompany single action potentials. In upper-layer cortical neurons, we were able to detect transients associated with single action potentials firing at rates of <1 Hz, with high reliability, from in vivo recordings in living mice.
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Max Planck Institute for Biological Cybernetics, Spemannstra
e 41, 72076 Tübingen, Germany. -
Max Planck Institute for Medical Research, Jahnstra 29, 69120 Heidelberg, Germany.
- University of Göttingen Medical School, Waldweg 33, 37073 Göttingen, Germany.
- Department of Pharmacology, Howard Hughes Medical Institute, University of California at San Diego, 9500 Gilman Drive, La Jolla, California 92093, USA.
- Max Planck Institute of Neurobiology, Am Klopferspitz 18, 82152 München-Martinsried, Germany.
- These authors contributed equally to this work.
Correspondence to: Mazahir T Hasan2,5 e-mail: mhasan@mpimf-heidelberg.mpg.de
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