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. 2020 Jun 9;31(10):107735. doi: 10.1016/j.celrep.2020.107735

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© 2020 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

The Mobility of Desensitized KARs Affects the Kinetics of Mixed AMPAR/KAR-Mediated Currents

(A) Left: scatter dot plot of decay time constants (τ) of eEPSCs mediated by AMPA and mixed AMPA/LiGluK2 receptors in neurons expressing only endogenous AMPARs (untransfected, black) or expressing LiGluK2 (blue) or LiGluK2Δ16 (green). Right: representative averaged traces of eEPSCs mediated by AMPARs, AMPA/LiGluK2 receptors, and AMPA/LiGluK2Δ16 receptors. Note that for visualization purposes, the stimulation artifacts of black and green traces have been omitted.

(B) Scatter dot plot of decay time constants (τ) of eEPSC mediated by LiGluK2 (blue) or LiGluK2Δ16 receptors (green) recorded in the presence of GYKI 53655 (left) and corresponding normalized representative traces (right).

(C) Top: representative AMPA/LiGluK2 receptors-mediated mixed eEPSCs recorded before and after the delivery of the desensitizing train. Bottom: magnification of the framed areas showing AMPA/LiGluK2 receptors-mediated mixed eEPSC before, during, and after the train. Inset: superimposed normalized slow component of mixed eEPSC decay before (black) and after (blue) the train, showing acceleration in the “after” current.

(D) Left: weighted time constant (τ_weighted) of mixed eEPSC before and after the train. Right: relative weight of the AMPAR and LiGluK2 components in AMPA/LiGluK2 receptors-mediated mixed eEPSCs before and after the train. Please note that after the desensitizing train, current decay kinetics was accelerated, the AMPAR-mediated component increased, and the KAR-mediated component decreased.

(E) Top: representative traces of AMPA/LiGluK2Δ16 receptors-mediated mixed eEPSC before and after the desensitizing train. Bottom: magnification as in (C). Inset: superimposed normalized slow component of mixed AMPA/LiGluK2Δ16 eEPSC before (black) and after (green) the train.

(F) Left: weighted time constant (τ_weighted) of AMPA/LiGluK2Δ16 mixed eEPSC before and after the train, indicating unchanged current decay kinetics. Right: unaffected relative weight of AMPAR and KAR components in AMPA/LiGluK2Δ16 mixed eEPSCs before and after the train.

Data are presented as mean ± SEM, ^∗∗∗p < 0.001; ns, non-significant. See also Figure S4.