ZFIN ID: ZDB-PUB-170622-19
An E3-ligase-based method for ablating inhibitory synapses
Gross, G.G., Straub, C., Perez-Sanchez, J., Dempsey, W.P., Junge, J.A., Roberts, R.W., Trinh, l.e. .A., Fraser, S.E., De Koninck, Y., De Koninck, P., Sabatini, B.L., Arnold, D.B.
Date: 2016
Source: Nature Methods   13: 673-8 (Journal)
Registered Authors: Dempsey, William, Fraser, Scott E., Roberts, Richard, Trinh, Le
Keywords: none
MeSH Terms:
  • Animals
  • Carrier Proteins/metabolism*
  • Cells, Cultured
  • Embryo, Mammalian/cytology
  • Embryo, Mammalian/metabolism
  • Female
  • Hippocampus
  • Male
  • Membrane Proteins/metabolism*
  • Motor Disorders/metabolism
  • Motor Disorders/pathology
  • Neurons/cytology
  • Neurons/metabolism*
  • Patch-Clamp Techniques/methods*
  • Rats
  • Rats, Sprague-Dawley
  • Spine/cytology
  • Spine/metabolism
  • Synapses/physiology*
  • Synaptic Transmission/physiology*
  • Ubiquitin-Protein Ligases/metabolism*
  • Ubiquitination
  • Zebrafish
PubMed: 27271196 Full text @ Nat. Methods
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.