title: Fluorescent protein voltage probes derived from ArcLight that respond to membrane voltage changes with fast kinetics.
abstract: We previously reported the discovery of a fluorescent protein voltage probe, ArcLight, and its derivatives that exhibit large changes in fluorescence intensity in response to changes of plasma membrane voltage. ArcLight allows the reliable detection of single action potentials and sub-threshold activities in individual neurons and dendrites. The response kinetics of ArcLight (τ1-on ~10 ms, τ2-on ~ 50 ms) are comparable with most published genetically-encoded voltage probes. However, probes using voltage-sensing domains other than that from the Ciona intestinalis voltage sensitive phosphatase exhibit faster kinetics. Here we report new versions of ArcLight, in which the Ciona voltage-sensing domain was replaced with those from chicken, zebrafish, frog, mouse or human. We found that the chicken and zebrafish-based ArcLight exhibit faster kinetics, with a time constant (τ) less than 6 ms for a 100 mV depolarization. Although the response amplitude of these two probes (8-9%) is not as large as the Ciona-based ArcLight (~35%), they are better at reporting action potentials from cultured neurons at higher frequency. In contrast, probes based on frog, mouse and human voltage sensing domains were either slower than the Ciona-based ArcLight or had very small signals.
authors: Cellular and Molecular Physiology, Yale University School of Medicine, New Haven, Connecticut, United States of America ; The John B. Pierce Laboratory, Inc., New Haven, Connecticut, United States of America.
authors: Zhou
authors: Z
authors: Han
authors: Lei
authors: L
authors: Jin
authors: Jelena
authors: J
authors: Platisa
authors: Lawrence B
authors: LB
authors: Cohen
authors: Bradley J
authors: BJ
authors: Baker
authors: Vincent A
authors: VA
authors: Pieribone
mesh_terms: D000200:Action Potentials; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D007700:Kinetics; D008164:Luminescent Proteins / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D009068:Movement; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism*; D013045:Species Specificity
chemical_list: C576393:Arclight fusion protein; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins