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

TitleAn E3-ligase-based method for ablating inhibitory synapses.
Publication TypeJournal Article
Year of Publication2016
AuthorsGross, GG, Straub, C, Perez-Sanchez, J, Dempsey, WP, Junge, JA, Roberts, RW, Trinh, LA, Fraser, SE, De Koninck, Y, De Koninck, P, Sabatini, BL, Arnold, DB
JournalNat Methods
Date Published2016 Aug
KeywordsAnimals, 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

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.

Alternate JournalNat. Methods
PubMed ID27271196
PubMed Central IDPMC5312699
Grant ListR01 AI085583 / AI / NIAID NIH HHS / United States
R01 NS081678 / NS / NINDS NIH HHS / United States
R01 GM083898 / GM / NIGMS NIH HHS / United States
R01 NS081687 / NS / NINDS NIH HHS / United States
R01 NS046579 / NS / NINDS NIH HHS / United States