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Figure 5. Diazepam modulation of GABA-evoked currents from α1β3 GABAA receptors.Xenopus laevis oocytes were injected with cRNA and subjected to two-electrode voltage clamp electrophysiology as described in the methods. Control currents (Icontrol) were evoked using a GABA concentration corresponding to ~EC5–10 and modulation by diazepam was evaluated by co-applications with GABAcontrol. (A,B) Representative GABA-evoked current traces from oocytes injected with the denoted cRNA mixtures. Bars above each trace indicate GABA, diazepam (Diaz) and flumazenil (Flu) concentrations and application periods. Dotted lines indicate the peak current amplitude by GABAcontrol and “/” a wash period. (C) Concentration-response relationships of diazepam modulation of GABAcontrol-evoked currents at α1β3 receptors stemming from injecting the indicated cRNA mixtures using free subunits or a concatenated β3-α1 construct. Averaged modulatory values were depicted as means ± S.E as a function of the diazepam concentration and fitted to the Hill equation by non-linear regression. Each data point represents experiments from n = 6 oocytes from ≥2 batches. Regression results with 95% confidence intervals for α1 + β3 (30:1) were: EC50 value of 0.040 μM (0.010–0.12) and Emax value of 40% (33–48) whereas results for β3-α1 + α1 (1:2) were: EC50 value of 0.020 μM (0.010–0.04) and Emax value of 51% (46–56). Image published in: Che Has AT et al. (2016) Copyright © 2016, Macmillan Publishers Limited. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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