An E3-ligase-based method for ablating inhibitory synapses

Nat Methods. 2016 Aug;13(8):673-8. doi: 10.1038/nmeth.3894. Epub 2016 Jun 6.

Abstract

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

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

Substances

  • Carrier Proteins
  • Membrane Proteins
  • gephyrin
  • Ubiquitin-Protein Ligases