Article abstract
Nature Methods 5, 805 - 811 (2008)
Published online: 10 August 2008 | doi:10.1038/nmeth.1243
A genetically encoded calcium indicator for chronic in vivo two-photon imaging
Marco Mank1, Alexandre Ferrão Santos1, Stephan Direnberger1, Thomas D Mrsic-Flogel1,3, Sonja B Hofer1,3, Valentin Stein1, Thomas Hendel1, Dierk F Reiff1, Christiaan Levelt2, Alexander Borst1, Tobias Bonhoeffer1, Mark Hübener1 & Oliver Griesbeck1
Abstract
Neurons in the nervous system can change their functional properties over time. At present, there are no techniques that allow reliable monitoring of changes within identified neurons over repeated experimental sessions. We increased the signal strength of troponin C–based calcium biosensors in the low-calcium regime by mutagenesis and domain rearrangement within the troponin C calcium binding moiety to generate the indicator TN-XXL. Using in vivo two-photon ratiometric imaging, we show that TN-XXL exhibits enhanced fluorescence changes in neurons of flies and mice. TN-XXL could be used to obtain tuning curves of orientation-selective neurons in mouse visual cortex measured repeatedly over days and weeks. Thus, the genetically encoded calcium indicator TN-XXL allows repeated imaging of response properties from individual, identified neurons in vivo, which will be crucial for gaining new insights into cellular mechanisms of plasticity, regeneration and disease.
- Max Planck Institute of Neurobiology, Am Klopferspitz 18, D-82152 Martinsried, Germany.
- Netherlands Institute for Neuroscience, Royal Netherlands Academy for Arts and Sciences, Meibergdreef 471105 BA, Amsterdam, The Netherlands.
- Present address: Department of Physiology, University College London, London WC1E 6JJ, UK.
Correspondence to: Oliver Griesbeck1 e-mail: griesbeck@neuro.mpg.de
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