2D measurement of ion currents associated to the signal transduction of the phototactic alga Chlamydomonas reinhardtii

Our objective was to develop a simple procedure for the detection of light-induced ion currents of photomotile cells in two dimensions. The novel technique was based on the light gradient method (LGM), and the model object was Chlamydomonas reinhardtii, a phototactic unicellular alga, ideal for such experiments. The conventional LGM cuvette was modified such that the electrode pair could be rotated around the sample and pick up the electric signals from arbitrary directions. The experiments were performed with and without the application of an auxiliary light beam preorienting the motile cells. The analysis of the detected traces revealed two main vectorial components of the signal by the help of singular value decomposition (SVD), in concert with previous experimental findings and theoretical considerations suggesting different origins of the “fast” and “slow” components of the photoelectric response of Chlamydomonas and Haematococcus cells. Using plausible assumptions, our method allowed a quantitative analysis of the signal, assigning size and direction to the two vectorial components. The method allows a rapid and accurate way to measure electric signals of photomotive cells in 2D, and particularly to test the physiological activity and in vivo-kinetics of site-directed mutants of ChR1 or ChR2, providing novel photo-electrophysiological methods with important quantitative information.