Fast and sensitive GCaMP calcium indicators for imaging neural populations
Looger, L.; Zhang, Y.; Rozsa, M.; Liang, Y.; Bushey, D.; Wei, Z.; Zheng, J.; Reep, D.; Broussard, G. J.; Tsang, A.; Tsegaye, G.; Narayan, S.; Obara, C. J.; Lim, J.-X.; Patel, R.; Zhang, R.; Ahrens, M. B.; Turner, G. C.; Wang, S. S.; Korff, W.; Schreiter, E. R.; Svoboda, K.; Hasseman, J. P.; Kolb, I.
Show abstract
Calcium imaging with protein-based indicators is widely used to follow neural activity in intact nervous systems. The popular GCaMP indicators are based on the calcium-binding protein calmodulin and the RS20 peptide. These sensors report neural activity at timescales much slower than electrical signaling, limited by their biophysical properties and trade-offs between sensitivity and speed. We used large-scale screening and structure-guided mutagenesis to develop and optimize several fast and sensitive GCaMP-type indicators. The resulting jGCaMP8 sensors, based on calmodulin and a fragment of endothelial nitric oxide synthase, have ultra-fast kinetics (rise times, 2 ms) and still feature the highest sensitivity for neural activity reported for any protein-based sensor. jGCaMP8 sensors will allow tracking of larger populations of neurons on timescales relevant to neural computation.
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