Combining Voltage and Calcium Imaging from Neuronal Dendrites |
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Authors: | Marco Canepari Kaspar Vogt Dejan Zecevic |
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Affiliation: | (1) Biozentrum, Department of Pharmacology and Neurobiology, University of Basel, Klingelbergstrasse 70, 4056 Basel, Switzerland;(2) Department of Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, CT 06520, USA |
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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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Keywords: | Voltage-sensitive dyes Calcium-sensitive dyes Imaging Neuronal dendrites |
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