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An E3-ligase-based method for ablating inhibitory synapses.

Publication Type:

Journal Article

Source:

Nat Methods, Volume 13, Issue 8, p.673-8 (2016)

Keywords:

Animals, Carrier Proteins, Cells, Cultured, Embryo, Mammalian, Female, Hippocampus, Male, Membrane Proteins, Motor Disorders, Neurons, Patch-Clamp Techniques, Rats, Rats, Sprague-Dawley, Spine, Synapses, Synaptic Transmission, Ubiquitin-Protein Ligases, Ubiquitination, Zebrafish

Abstract:

<p>Although neuronal activity can be modulated using a variety of techniques, there are currently few methods for controlling neuronal connectivity. We introduce a tool (GFE3) that mediates the fast, specific and reversible elimination of inhibitory synaptic inputs onto genetically determined neurons. GFE3 is a fusion between an E3 ligase, which mediates the ubiquitination and rapid degradation of proteins, and a recombinant, antibody-like protein (FingR) that binds to gephyrin. Expression of GFE3 leads to a strong and specific reduction of gephyrin in culture or in vivo and to a substantial decrease in phasic inhibition onto cells that express GFE3. By temporarily expressing GFE3 we showed that inhibitory synapses regrow following ablation. Thus, we have created a simple, reversible method for modulating inhibitory synaptic input onto genetically determined cells.</p>

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