PUBLICATION
An E3-ligase-based method for ablating inhibitory synapses
- Authors
- 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.
- ID
- ZDB-PUB-170622-19
- 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
Citation
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. (2016) An E3-ligase-based method for ablating inhibitory synapses. Nature Methods. 13:673-8.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping