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. 2020 Mar 16;10:4804. doi: 10.1038/s41598-020-61677-w

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© The Author(s) 2020

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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Functional inhibition of wild-type α3GlyRs by cAMP signaling. (A) Current traces from cells expressing α3GlyRs alone or together with EP2 receptors or with bPAC. Sample traces were obtained after 0, 1 and 10 minutes (‘) of recording. (B) Time course of glycine-evoked currents (500 μM) in the absence and presence of blue light in cells co-expressing α3GlyRs and bPAC. (C) Summary of the glycine-activated current inhibition elicited by EP2 receptor activation with PGE₂ (n = 4, red) or by bPAC stimulation with blue light (n = 11, blue) after 10 minutes of recording. Control conditions (PGE₂ without the expression of EP2 receptors (n = 5, white), +bPAC under no light conditions (n = 10, black) and cells stimulated with blue light without bPAC expressed (n = 5, cyan) are also shown. Differences were significant between the groups treated with PGE₂ (*P < 0.05) and between the groups expressing bPAC (***P < 0.001). ANOVA followed by Bonferroni post-hoc test, F(4, 34) = 8.99. (D) Plasma membrane expression of α3GlyRs after EP2 receptor activation or after bPAC stimulation with blue light. Calibration bar, 5 μm. (E) The graph shows the quantification of the receptor-associated signals (left) and number of receptor clusters (right). Differences were not significant, (Fluorescence intensity, F(3, 51) = 0.25; number of receptor clusters, F(3, 31) = 1.1). Relative fluorescence intensity: control (vehicle), n = 13; no light, n = 13; EP2 + PGE2, n = 12; blue light, n = 14. Number of clusters: control (vehicle), n = 8; no light, n = 8; EP2 + PGE2, n = 8; blue light, n = 8.