An E3-ligase-based method for ablating inhibitory synapses.
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.
Department of Biology, Section of Molecular and Computational Biology, University of Southern California, Los Angeles, Los Angeles, California, USA.
Garrett G
GG
Gross
Howard Hughes Medical Institute, Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA.
Christoph
C
Straub
Centre de recherche, Institut universitaire en santé mentale de Québec, Québec, Québec, Canada.|Université Laval, Québec, Québec, Canada.
Jimena
J
Perez-Sanchez
Department of Biology, Section of Molecular and Computational Biology, University of Southern California, Los Angeles, Los Angeles, California, USA.
William P
WP
Dempsey
Department of Biology, Section of Molecular and Computational Biology, University of Southern California, Los Angeles, Los Angeles, California, USA.
Jason A
JA
Junge
Department of Chemistry, University of Southern California, Los Angeles, Los Angeles, California, USA.
Richard W
RW
Roberts
Department of Biology, Section of Molecular and Computational Biology, University of Southern California, Los Angeles, Los Angeles, California, USA.
Le A
le A
Trinh
Department of Biology, Section of Molecular and Computational Biology, University of Southern California, Los Angeles, Los Angeles, California, USA.
Scott E
SE
Fraser
Centre de recherche, Institut universitaire en santé mentale de Québec, Québec, Québec, Canada.|Université Laval, Québec, Québec, Canada.
Yves
0000-0002-5779-9330
Y
De Koninck
Centre de recherche, Institut universitaire en santé mentale de Québec, Québec, Québec, Canada.|Université Laval, Québec, Québec, Canada.
Paul
P
De Koninck
Howard Hughes Medical Institute, Department of Neurobiology, Harvard Medical School, Boston, Massachusetts, USA.
Bernardo L
BL
Sabatini
Department of Biology, Section of Molecular and Computational Biology, University of Southern California, Los Angeles, Los Angeles, California, USA.
Don B
DB
Arnold
D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism; D005260:Female; D006624:Hippocampus; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D000068079:Motor Disorders / Q000378:metabolism / Q000473:pathology; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D018408:Patch-Clamp Techniques / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013131:Spine / Q000166:cytology / Q000378:metabolism; D013569:Synapses / Q000502:physiology*; D009435:Synaptic Transmission / Q000502:physiology*; D044767:Ubiquitin-Protein Ligases / Q000378:metabolism*; D054875:Ubiquitination; D015027:Zebrafish
D002352:Carrier Proteins; D008565:Membrane Proteins; C073441:gephyrin; D044767:Ubiquitin-Protein Ligases