Calcium-permeable channelrhodopsins for the photocontrol of calcium signalling.

Autor: Fernandez Lahore, Rodrigo G., Pampaloni, Niccolò P., Peter, Enrico, Heim, M.-Marcel, Tillert, Linda, Vierock, Johannes, Oppermann, Johannes, Walther, Jakob, Schmitz, Dietmar, Owald, David, Plested, Andrew J. R., Rost, Benjamin R., Hegemann, Peter
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Zdroj: Nature Communications; 12/21/2022, Vol. 13 Issue 1, p1-18, 18p
Abstrakt: Channelrhodopsins are light-gated ion channels used to control excitability of designated cells in large networks with high spatiotemporal resolution. While ChRs selective for H+, Na+, K+ and anions have been discovered or engineered, Ca2+-selective ChRs have not been reported to date. Here, we analyse ChRs and mutant derivatives with regard to their Ca2+ permeability and improve their Ca2+ affinity by targeted mutagenesis at the central selectivity filter. The engineered channels, termed CapChR1 and CapChR2 for calcium-permeable channelrhodopsins, exhibit reduced sodium and proton conductance in connection with strongly improved Ca2+ permeation at negative voltage and low extracellular Ca2+ concentrations. In cultured cells and neurons, CapChR2 reliably increases intracellular Ca2+ concentrations. Moreover, CapChR2 can robustly trigger Ca2+ signalling in hippocampal neurons. When expressed together with genetically encoded Ca2+ indicators in Drosophila melanogaster mushroom body output neurons, CapChRs mediate light-evoked Ca2+ entry in brain explants. To date, no Ca2 + -selective channelrhodopsins have been characterized. In this study, Fernandez Lahore et al. report two calcium-permeable channelrhodopsins (CapChR1 and 2) for the photocontrol of calcium signalling in excitable tissue. [ABSTRACT FROM AUTHOR]
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