Red-shifted voltage-sensitive fluorescent proteins.
Electrical signals generated by nerve cells provide the basis of brain function. Whereas single or small numbers of cells are easily accessible using microelectrode recording techniques, less invasive optogenetic methods with spectral properties optimized for in vivo imaging are required for elucidating the operation mechanisms of neuronal circuits composed of large numbers of neurons originating from heterogeneous populations. To this end, we generated and characterized a series of genetically encoded voltage-sensitive fluorescent proteins by molecular fusion of the voltage-sensing domain of Ci-VSP (Ciona intestinalis voltage sensor-containing phosphatase) to red-shifted fluorescent protein operands. We show how these indicator proteins convert voltage-dependent structural rearrangements into a modulation of fluorescence output and demonstrate their applicability for optical recording of individual or simultaneous electrical signals in cultured hippocampal neurons at single-cell resolution without temporal averaging.
Perron
Amelie
A
Laboratory for Neuronal Circuit Dynamics, RIKEN Brain Science Institute, Wako-shi, Saitama 351-0198, Japan.
Mutoh
Hiroki
H
Launey
Thomas
T
Knöpfel
Thomas
T
D000818:Animals; D045744:Cell Line, Tumor; D055724:Electrophysiological Phenomena; D007700:Kinetics; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism*; D008839:Microelectrodes; D009474:Neurons / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000235:genetics / Q000378:metabolism; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D000097573:Red Fluorescent Protein
D008164:Luminescent Proteins; D010744:Phosphoric Monoester Hydrolases; D011993:Recombinant Fusion Proteins; D000097573:Red Fluorescent Protein; C509671:voltage-sensor-containing phosphatase, Ciona intestinalis