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Sci Rep ; 6: 23976, 2016 Apr 07.
Article in English | MEDLINE | ID: mdl-27052670

ABSTRACT

Growth cones of extending axons navigate to correct targets by sensing a guidance cue gradient via membrane protein receptors. Although most signaling mechanisms have been clarified using an in vitro approach, it is still difficult to investigate the growth cone behavior in complicated extracellular environment of living animals due to the lack of tools. We develop a system for the light-dependent activation of a guidance receptor, Deleted in Colorectal Cancer (DCC), using Arabidopsis thaliana Cryptochrome 2, which oligomerizes upon blue-light absorption. Blue-light illumination transiently activates DCC via its oligomerization, which initiates downstream signaling in the illuminated subcellular region. The extending axons are attracted by illumination in cultured chick dorsal root ganglion neurons. Moreover, light-mediated navigation of the growth cones is achieved in living Caenorhabditis elegans. The photo-manipulation system is applicable to investigate the relationship between the growth cone behavior and its surrounding environment in living tissue.


Subject(s)
Axon Guidance/physiology , Axons/physiology , Neuronal Outgrowth/physiology , Optogenetics/methods , Receptors, Cell Surface/metabolism , Animals , Animals, Genetically Modified , Axon Guidance/radiation effects , Axons/metabolism , Axons/radiation effects , Caenorhabditis elegans/genetics , Caenorhabditis elegans/metabolism , Caenorhabditis elegans/radiation effects , Chick Embryo , Chickens , Ganglia, Spinal/cytology , Ganglia, Spinal/embryology , Ganglia, Spinal/metabolism , HEK293 Cells , Humans , Immunoblotting , Light , Mice , Microscopy, Fluorescence , Neuronal Outgrowth/radiation effects , Neurons/metabolism , Neurons/physiology , Neurons/radiation effects , Receptors, Cell Surface/genetics , Tumor Suppressor Proteins/genetics , Tumor Suppressor Proteins/metabolism
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