title: Rapid and permanent neuronal inactivation in vivo via subcellular generation of reactive oxygen with the use of KillerRed.
abstract: Inactivation of selected neurons in vivo can define their contribution to specific developmental outcomes, circuit functions, and behaviors. Here, we show that the optogenetic tool KillerRed selectively, rapidly, and permanently inactivates different classes of neurons in C. elegans in response to a single light stimulus, through the generation of reactive oxygen species (ROS). Ablation scales from individual neurons in single animals to multiple neurons in populations and can be applied to freely behaving animals. Using spatially restricted illumination, we demonstrate that localized KillerRed activation in either the cell body or the axon triggers neuronal degeneration and death of the targeted cell. Finally, targeting KillerRed to mitochondria results in organelle fragmentation without killing the cell, in contrast to the cell death observed when KillerRed is targeted to the plasma membrane. We expect this genetic tool to have wide-ranging applications in studies of circuit function and subcellular responses to ROS.
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Williams
authors.forename: Rachid El
authors.initials: RE
authors.lastname: Bejjani
authors.forename: Paula Mugno
authors.initials: PM
authors.lastname: Ramirez
authors.forename: Sean
authors.initials: S
authors.lastname: Coakley
authors.forename: Shin Ae
authors.initials: SA
authors.lastname: Kim
authors.forename: Hyewon
authors.initials: H
authors.lastname: Lee
authors.forename: Quan
authors.initials: Q
authors.lastname: Wen
authors.forename: Aravi
authors.initials: A
authors.lastname: Samuel
authors.forename: Hang
authors.initials: H
authors.lastname: Lu
authors.forename: Massimo A
authors.initials: MA
authors.lastname: Hilliard
authors.forename: Marc
authors.initials: M
authors.lastname: Hammarlund
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000528:radiation effects; D017173:Caenorhabditis elegans / Q000378:metabolism; D059330:GABAergic Neurons / Q000187:drug effects / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D008027:Light; D018715:Microscopy, Video; D017382:Reactive Oxygen Species / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000378:metabolism
chemical_list: D017382:Reactive Oxygen Species; C508771:killer red protein, Anthomedusae; D049452:Green Fluorescent Proteins; D013482:Superoxide Dismutase