Abstract
The ability to monitor membrane potential (V m) and calcium (Ca2+) transients at multiple locations on the same neuron can facilitate further progress in our understanding of neuronal function. Here we describe a method to combine V m and Ca2+ imaging using styryl voltage sensitive dyes and Fura type UV-excitable Ca2+ indicators. In all cases V m optical signals are linear with membrane potential changes, but the calibration of optical signals on an absolute scale is presently possible only in some neurons. The interpretation of Ca2+ optical signals depends on the indicator Ca2+ buffering capacity relative to the cell endogenous buffering capacity. In hippocampal CA1 pyramidal neurons, loaded with JPW-3028 and 300 μM Bis-Fura-2, V m optical signals cannot be calibrated and the physiological Ca2+ dynamics are compromised by the presence of the indicator. Nevertheless, at each individual site, relative changes in V m and Ca2+ fluorescence signals under different conditions can provide meaningful new information on local dendritic integration. In cerebellar Purkinje neurons, loaded with JPW-1114 and 1 mM Fura-FF, V m optical signals can be calibrated in terms of mV and Ca2+ optical signals quantitatively reveal the physiological changes in free Ca2+. Using these two examples, the method is explained in detail.
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Acknowledgements
This work was supported by the University of Basel and the NIH Grant RO1NS42739. We are grateful to Leslie Loew and Joe Wuskel for kindly providing voltage-sensitive dyes, and to Helene Pierre for technical help.
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Canepari, M., Vogt, K. & Zecevic, D. Combining Voltage and Calcium Imaging from Neuronal Dendrites. Cell Mol Neurobiol 28, 1079–1093 (2008). https://doi.org/10.1007/s10571-008-9285-y
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DOI: https://doi.org/10.1007/s10571-008-9285-y