Abstract
The role the cytoskeleton plays in generating and/or maintaining gephyrin-dependent receptor clusters at inhibitory synapses is poorly understood. Here, the effects of actin cytoskeleton disruption were investigated in eGFP-gephyrin-transfected cells and hippocampal neurons. While gephyrin was not associated with microfilaments in transfected cells, it colocalized with G-actin and cytochalasin-D-induced F-actin patches. The linker region between the MoeA and MogA homology domains of gephyrin was required for colocalization with F-actin patches and for the binding of gephyrin to ena/VASP, an actin anti-capping factor that, in vitro, caused gephyrin binding to polymerized actin. In hippocampal neurons, treatment with cytochalasin D resulted in the redistribution of the neuronal ena/VASP homologue Mena into actin patches and, at early stages of development, a reduction in the number of gephyrin clusters. Our data suggest that Mena binding to F-actin allows for gephyrin recruitment to the leading edge of uncapped actin filaments.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Actin Cytoskeleton / metabolism
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Actins / genetics
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Actins / metabolism*
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Animals
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Antineoplastic Agents / pharmacology
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Bridged Bicyclo Compounds, Heterocyclic / pharmacology
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Carrier Proteins / genetics
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Carrier Proteins / metabolism*
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Cell Adhesion Molecules / genetics
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Cell Adhesion Molecules / metabolism*
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Cells, Cultured
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Cytochalasin D / pharmacology
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Cytoskeleton / drug effects
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Cytoskeleton / metabolism*
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Depsipeptides / pharmacology
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Hippocampus / cytology
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Humans
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Membrane Proteins / genetics
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Membrane Proteins / metabolism*
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Microfilament Proteins / genetics
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Microfilament Proteins / metabolism*
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Neurons / cytology
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Neurons / metabolism
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Nucleic Acid Synthesis Inhibitors / pharmacology
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Phosphoproteins / genetics
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Phosphoproteins / metabolism*
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Protein Structure, Tertiary
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Rats
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Synapses / metabolism
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Synapses / ultrastructure
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Thiazoles / pharmacology
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Thiazolidines
Substances
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Actins
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Antineoplastic Agents
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Bridged Bicyclo Compounds, Heterocyclic
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Carrier Proteins
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Cell Adhesion Molecules
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Depsipeptides
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Membrane Proteins
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Microfilament Proteins
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Nucleic Acid Synthesis Inhibitors
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Phosphoproteins
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Recombinant Fusion Proteins
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Thiazoles
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Thiazolidines
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gephyrin
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vasodilator-stimulated phosphoprotein
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jasplakinolide
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Cytochalasin D
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latrunculin A