An E3-ligase-based method for ablating inhibitory synapses

作者：Gross Garrett G; Straub Christoph; Perez Sanchez Jimena; Dempsey William P; Junge Jason A; Roberts Richard W; Trinh Le A; Fraser Scott E; De Koninck Yves; De Koninck Paul; Sabatini Bernardo L; Arnold Don B^*

来源：Nature Methods, 2016, 13(8): 673-+.

DOI：10.1038/nmeth.3894

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.

• 出版日期2016-8