A kainate receptor subunit promotes the recycling of the neuron-specific K+-Cl- co-transporter KCC2 in hippocampal neurons

J Biol Chem. 2017 Apr 14;292(15):6190-6201. doi: 10.1074/jbc.M116.767236. Epub 2017 Feb 24.


Synaptic inhibition depends on a transmembrane gradient of chloride, which is set by the neuron-specific K+-Cl- co-transporter KCC2. Reduced KCC2 levels in the neuronal membrane contribute to the generation of epilepsy, neuropathic pain, and autism spectrum disorders; thus, it is important to characterize the mechanisms regulating KCC2 expression. In the present study, we determined the role of KCC2-protein interactions in regulating total and surface membrane KCC2 expression. Using quantitative immunofluorescence in cultured mouse hippocampal neurons, we discovered that the kainate receptor subunit GluK2 and the auxiliary subunit Neto2 significantly increase the total KCC2 abundance in neurons but that GluK2 exclusively increases the abundance of KCC2 in the surface membrane. Using a live cell imaging assay, we further determined that KCC2 recycling primarily occurs within 1-2 h and that GluK2 produces an ∼40% increase in the amount of KCC2 recycled to the membrane during this time period. This GluK2-mediated increase in surface recycling translated to a significant increase in KCC2 expression in the surface membrane. Moreover, we found that KCC2 recycling is enhanced by protein kinase C-mediated phosphorylation of the GluK2 C-terminal residues Ser-846 and Ser-868. Lastly, using gramicidin-perforated patch clamp recordings, we found that the GluK2-mediated increase in KCC2 recycling to the surface membrane translates to a hyperpolarization of the reversal potential for GABA (EGABA). In conclusion, our results have revealed a mechanism by which kainate receptors regulate KCC2 expression in the hippocampus.

Keywords: K+-Cl− co-transporter (KCC2); cation-chloride co-transporter (CCC); gamma-aminobutyric acid (GABA); glutamate receptor; membrane recycling; neuron; protein kinase C (PKC); protein phosphorylation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Membrane / genetics
  • Cell Membrane / metabolism*
  • Cells, Cultured
  • Hippocampus / cytology
  • Hippocampus / metabolism*
  • Membrane Potentials / physiology*
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Mice
  • Mice, Knockout
  • Neurons / cytology
  • Neurons / metabolism*
  • Receptors, Kainic Acid / genetics
  • Receptors, Kainic Acid / metabolism*
  • Symporters / genetics
  • Symporters / metabolism*


  • Gluk2 kainate receptor
  • Membrane Proteins
  • NETO2 protein, mouse
  • Receptors, Kainic Acid
  • Symporters
  • potassium-chloride symporters

Grant support