Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2015 Sep 3;6:8038. doi: 10.1038/ncomms9038

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Copyright © 2015, Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved.

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

PMC Copyright notice

(a–c) KCC2 mutants exhibit a depolarized E_Cl. (a) Examples of I–V relationships in HEK293 cells transfected either with GlyR α₂ alone (no KCC2), or GlyR α₂ with a wild-type KCC2 construct. I_Gly amplitude was plotted against the holding potential and the intercept of the line of fit of this relation with the abscissa was taken as E_Cl. Superimposed traces show examples of glycine-evoked currents recorded from individual cells at different holding potentials (–80 to +20 mV). (b) Examples of I–V relationships in HEK293 cells co-transfected with GlyR α₂ and mutant KCC2, as indicated. E_Cl in mutant KCC2 is depolarized in comparison to wild-type (fit of I–V relation indicated by dashed-line). Calibration bars for 3a and 3b: 1nA, 1sec. (c) Summary chart plotting the distribution of E_Cl for each cell group: no KCC2: –14.7±0.8 mV (n=5), GlyR-KCC2: –29±3.5 mV (n=9), G551D: –17.4±1.2 mV (n=5), L426P: –17.9±2.9 mV (n=5), L311H: –15±3.1 mV (n=9); *P=0.006, one-way analysis of variance (ANOVA). Grey symbols represent measurements from individual cells. Boxes represent the mean±standard error of the mean. GlyR α₂: glycine receptor α₂, Wt-KCC2: wild-type K⁺-Cl⁻ co-transporter. (d–f) The rate of Cl⁻ extrusion is impaired in KCC2 mutants. (d) I_Gly amplitude plotted against time in a HEK293 cell co-transfected with GlyRα₂ and wild-type KCC2 and held at different membrane potentials as indicated in the top image. Vertical bars indicate the times of successive glycine applications. Example traces show glycine-evoked currents at times indicated by letters in italic. Calibration bars: 1 nA, 300 ms. (e) Rate of recovery of I_Gly amplitude after switching back the holding potential from +40 to −80 mV. Fine lines replace symbols and error bars are omitted for legibility. An exponential decay function was used to fit I_Gly amplitude ratio in each cell group. (f) Summary bar graph of the time-constant of I_Gly recovery in each group (no KCC2: 224±56.7 s, n=6; WT-KCC2: 59.5±7.7 s, n=13; L426P: 162.5±42.1 s, n=4; G551D: 340.1±137 s, n=4; L311H: 88.1±11.5 s, n=7). *Different from WT-KCC2 (P<0.01), ANOVA.