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. 2017 Jun 20;7:3870. doi: 10.1038/s41598-017-04209-3

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Acute CLP257 restores Cl⁻ extrusion in SDH neurons of morphine-treated rats. (a–c) Voltage clamp responses at different holding potentials to 30-ms muscimol puffs (solid line) in SDH neurons following saline (a) or morphine (b) treatments in the presence of a Cl⁻ load (29 mM) to measure Cl⁻ extrusion capacity. In (c), muscimol response in a SDH neuron from a morphine-treated rat obtained after slice pre-incubation with CLP257 (100 µM) for 1 hour. The colored line indicates the response at −55.5 mV (note the change in polarity). (d) I-V relationships for GABA_A currents obtained from neurons in (a–c). (e) Pooled E _GABA in controls (n = 7 cells, blue), in morphine-treated rats (n = 7 cells, red) and following CLP257 treatment (n = 7 cells, orange). E _GABA is significantly more depolarized following morphine treatment as compared to control rats (one-way ANOVA, P = 0.02, Tukey post-hoc *P = 0.02), but not after CLP257 pre-incubation (P = 0.7); MOR vs. MOR + CLP (P = 0.1). Abbreviations: CTR = control; MOR = morphine; SDH = superficial dorsal horn.