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. 2017 Apr 6;7:702. doi: 10.1038/s41598-017-00771-y

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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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Effect of blocking gap junction permeability on GLYT1 activation. (a) Time course of GVBD in oocytes isolated from punctured antral follicles that had been cultured for the period indicated in the presence or absence of the gap junction blocker, AGA (150 µM). Means were significantly different for all time points from 2–10 h inclusive. (b) GLYT1 activation as a function of time in the presence or absence of AGA in the same oocytes as in (a). GLYT1 activity was significantly increased starting at 5 h, with partial activation at 5 h and full activation at 7.5 and 10 h. Each point represents the mean ± s.e.m. of three independent repeats at each time (in A and B). (c) Time course of GVBD in oocytes isolated from COCs cultured with or without AGA identically to the antral follicles in (a). Significant GVBD occurred starting at 1 h and was maximal by 2–3 h. (d) GLYT1 activity in the same oocytes as in (c). There was no significant difference in the presence vs. absence of AGA. Each point represents the mean ± s.e.m. of three independent repeats at each time (in c and d). (e) Time course of GVBD in oocytes isolated from punctured antral follicles in the presence of no addition (Control), Connexin Mimetic Peptide directed against Connexin 43 (Cx43 CMP) or a scrambled peptide. GVBD was significantly increased only by 10 h only in the Cx43 CMP treated group. (f) GLYT1 activity in the same oocytes as in (e) did not increase significantly until starting at 16 h only in the Cx43 CMP group. Each point represents the mean ± s.e.m. of three independent repeats at each time (in e and f). Throughout (a–f), data were analysed by 2-way ANOVA (overall effect between treatments: P < 0.0001 for a,b,c,e,f; P = 0.18 for d) with Bonferroni post-tests between treatments at individual time points (*P < 0.05; ***P < 0.001; NS not significant: P > 0.05).