Optimization of a GCaMP Calcium Indicator for Neural Activity Imaging

Genetically encoded calcium indicators (GECIs) are powerful tools for systems neuroscience. Recent efforts in protein engineering have significantly increased the performance of GECIs. The state-of-the art single-wavelength GECI, GCaMP3, has been deployed in a number of model organisms and can reliably detect three or more action potentials in short bursts in several systems in vivo. Through protein structure determination, targeted mutagenesis, high-throughput screening, and a battery of in vitro assays, we have increased the dynamic range of GCaMP3 by severalfold, creating a family of "GCaMP5" sensors. We tested GCaMP5s in several systems: cultured neurons and astrocytes, mouse retina, and in vivo in…

• UD
  U.S. Department of Energy

  Awards: AC02-06CH11357, Contract No. DE-AC02-06CH11357, DE-AC02, 06CH11357, No. DE-AC02-06CH11357, DE-AC02-06CH11357, DE-AC02-
• EL
  Eli Lilly and Company
• NI
  National Institutes of Health

  Awards: R01 NS045193, NS045193, DE-AC02-06CH11357, NIH R01
• OO
  Office of Science

  Awards: DE-AC02-06CH11357, Contract No. DE-AC02-06CH11357, DE-AC02, No. DE-AC02-06CH11357, 06CH11357, AC02-06CH11357
• BE
  Basic Energy Sciences

  Awards: DE-AC02, Contract No. DE-AC02-06CH11357, AC02-06CH11357, 06CH11357
• AN
  Argonne National Laboratory

  Awards: DE-AC02, No. DE-AC02-06CH11357, 06CH11357, AC02-06CH11357, Contract No. DE-AC02-06CH11357