Re-evaluation of functional changes in representative GABRA1 variants from each group defined by the ΔlogEC50 method (IMAGE)
Caption
(A) Representative traces of GABA-evoked currents in wild-type (black), F325L (red), T257R (red), V194F (green), P360H (green), F92L (light blue), and V207F (light blue) α1 GABAARs. Black bars indicate 1-second applications of 0.1–1000 μM GABA. A uniform scale bar is shown. (B) Three variants that shifted the dose–response curve to the left (red), three variants that shifted the dose–response curve to the right (light blue), and three variants that did not shift the dose–response curve (green) were selected for further analysis. (C) Bar graph showing the EC50 values of the chosen variants and WT GABAAR. (D) Peak amplitudes of each receptor were normalized to the maximum response of WT GABAAR. Variants in both the left-shifted (red) and non-shifted (green) groups exhibited different directional changes in response amplitude within the same group. (E) Bar graph showing the accumulated charges passing through the receptors across all GABA concentrations tested for the selected variants. The results demonstrated that there are only 3 real gain-of-function variants (N261I, F325L, and P360H) among the 9 selected variants. This is consistent with the bar graphs. (F, G) based on our newly proposed response ratio method, showing the MT/WT amplitude ratio (F) and MT/WT charge flow ratio (G) at the EC50 of the WT receptor. The data shown in the figures represent the mean ± SEM from multiple independent experiments.
Credit
Wenlin Chen, Yee Wah So, Catherine Wang, Yanwen Shen, Michelle Demos, Jun Mu, Tian Rui Zhang, Xiaobin Wu, Linda Huh, Yu Tian Wang, Lidong Liu
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CC BY-NC-ND