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Sci Signal. 2019 Oct 15;12(603). pii: eaaw9315. doi: 10.1126/scisignal.aaw9315.

Developmentally regulated KCC2 phosphorylation is essential for dynamic GABA-mediated inhibition and survival.

Author information

1
Department of Neurophysiology, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka 431-3192, Japan.
2
Institute of Biomedical and Clinical Sciences, Medical School, College of Medicine and Health, University of Exeter, Hatherly Laboratories, Exeter EX4 4PS, UK.
3
Department of Neurosurgery, Yale School of Medicine, New Haven, CT 06510, USA.
4
Human Aging Research Institute, School of Life Sciences, Nanchang University, Nanchang, Jiangxi 330031, China.
5
Department of Anesthesiology, Vanderbilt University School of Medicine, Nashville, TN 37232, USA.
6
Division of Nephrology, Department of Medicine, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA 02215, USA.
7
The Broad Institute of Harvard and the Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
8
Laboratory of Human Genetics and Genomics, The Rockefeller University, New York, NY, USA.
9
Department of Genetics, Yale University School of Medicine, New Haven, CT 06511, USA.
10
Department of Neurophysiology, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka 431-3192, Japan. kristopher.kahle@yale.edu axfukuda@hama-med.ac.jp.
11
Advanced Research Facilities and Services, Preeminent Medical Photonics Education and Research Center, Hamamatsu University School of Medicine, Hamamatsu, Shizuoka 431-3192, Japan.
12
Departments of Neurosurgery, Pediatrics, and Cellular and Molecular Physiology, Centers for Mendelian Genomics, Yale School of Medicine, New Haven, CT 06510, USA. kristopher.kahle@yale.edu axfukuda@hama-med.ac.jp.

Abstract

Despite its importance for γ-aminobutyric acid (GABA) inhibition and involvement in neurodevelopmental disease, the regulatory mechanisms of the K+/Cl- cotransporter KCC2 (encoded by SLC12A5) during maturation of the central nervous system (CNS) are not entirely understood. Here, we applied quantitative phosphoproteomics to systematically map sites of KCC2 phosphorylation during CNS development in the mouse. KCC2 phosphorylation at Thr906 and Thr1007, which inhibits KCC2 activity, underwent dephosphorylation in parallel with the GABA excitatory-inhibitory sequence in vivo. Knockin mice expressing the homozygous phosphomimetic KCC2 mutations T906E/T1007E (Kcc2E/E ), which prevented the normal developmentally regulated dephosphorylation of these sites, exhibited early postnatal death from respiratory arrest and a marked absence of cervical spinal neuron respiratory discharges. Kcc2E/E mice also displayed disrupted lumbar spinal neuron locomotor rhythmogenesis and touch-evoked status epilepticus associated with markedly impaired KCC2-dependent Cl- extrusion. These data identify a previously unknown phosphorylation-dependent KCC2 regulatory mechanism during CNS development that is essential for dynamic GABA-mediated inhibition and survival.

PMID:
31615901
DOI:
10.1126/scisignal.aaw9315

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