Imaging Fast Calcium Currents beyond the Limitations of Electrode Techniques

International audienceThe present understanding of Ca2+ channel function is derived from the use of the patch clamp technique. In particular, the measurement of fast cellular Ca2+ currents is routinely achieved using whole-cell voltage clamp recordings. This experimental approach, however, is not applicable to the study of local native Ca2+ channels during physiological changes of membrane potential in complex cells, since the voltage clamp configuration constrains the membrane potential to a given value. Here, we report for the first time that Ca2+ currents from individual cells can be quantitatively measured beyond the limitations of the voltage clamp approach using fast Ca2+ imaging with low-affinity indicators. The optical measurement of the Ca2+ current was correlated with the membrane potential, simultaneously measured with a voltage-sensitive dye to investigate the activation of Ca2+ channels along the apical dendrite of the CA1 hippocampal pyramidal neuron during the back-propagation of an action potential. To validate the method, we analysed the voltage-dependence of high- and low-voltage gated Ca2+ channels. In particular, we measured the Ca2+ current component mediated by T-type channels and we investigated the mechanisms of recovery from inactivation of these channels. This method is expected to become a reference approach to investigate Ca2+ channels in their native physiological environment