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Elife. 2017 Oct 13;6. pii: e28270. doi: 10.7554/eLife.28270.

Native KCC2 interactome reveals PACSIN1 as a critical regulator of synaptic inhibition.

Author information

1
Department of Cell and Systems Biology, University of Toronto, Toronto, Canada.
2
Department of Anatomy, Faculty of Medicine, University of Helsinki, Helsinki, Finland.
3
Department of Molecular Genetics, Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Canada.
4
Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto, Canada.
5
Institut Universitaire en Santé Mentale de Québec, Québec, Canada.
6
Department of Psychiatry and Neuroscience, Université Laval, Québec, Canada.
7
Queensland Brain Institute, Clem Jones Centre for Ageing Dementia Research, The University of Queensland, Brisbane, Australia.

Abstract

KCC2 is a neuron-specific K+-Cl- cotransporter essential for establishing the Cl- gradient required for hyperpolarizing inhibition in the central nervous system (CNS). KCC2 is highly localized to excitatory synapses where it regulates spine morphogenesis and AMPA receptor confinement. Aberrant KCC2 function contributes to human neurological disorders including epilepsy and neuropathic pain. Using functional proteomics, we identified the KCC2-interactome in the mouse brain to determine KCC2-protein interactions that regulate KCC2 function. Our analysis revealed that KCC2 interacts with diverse proteins, and its most predominant interactors play important roles in postsynaptic receptor recycling. The most abundant KCC2 interactor is a neuronal endocytic regulatory protein termed PACSIN1 (SYNDAPIN1). We verified the PACSIN1-KCC2 interaction biochemically and demonstrated that shRNA knockdown of PACSIN1 in hippocampal neurons increases KCC2 expression and hyperpolarizes the reversal potential for Cl-. Overall, our global native-KCC2 interactome and subsequent characterization revealed PACSIN1 as a novel and potent negative regulator of KCC2.

KEYWORDS:

Gaba; KCC2; PACSIN1; chloride; interactome; mouse; neuroscience; syndapin

PMID:
29028184
PMCID:
PMC5640428
DOI:
10.7554/eLife.28270
[Indexed for MEDLINE]
Free PMC Article

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