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. 2019 Dec 5;9:18395. doi: 10.1038/s41598-019-54693-y

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© The Author(s) 2019

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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Oocytes utilise AMP for ATP production via the adenosine salvage pathway. Iodoacetamide (IAC) was used to inhibit creatine kinase activity in order to perturb the interconversion of ADP and ATP. COCs (A) or denuded oocytes (B) were cultured for 3 h ± ¹³C₅-AMP and then intra-oocyte ¹³C₅-AMP, ¹³C₅-ADP and ¹³C₅-ATP were measured by LC-MS/MS (n = 4 biological replicates). COCs were cultured with ¹³C₅-AMP ± the gap-junction uncoupler carbenoxolone (CBX) for 3 h and intra-oocyte ¹³C₅-ATP was measured (n = 4 biological replicates) (C). COCs were cultured for 3 h with increasing concentrations of ¹³C₅-AMP and the peak areas of intra-oocyte ¹³C₅-ATP and endogenous ATP were measured (n = 1 biological replicate) (D). COCs were cultured for 3 h ± ¹³C₅-AMP (1 mM) and intra-oocyte ¹³C₅-ATP was measured (n = 4 biological replicates) (E). ns, not significantly different (P ˃ 0.05, t-test), ^#Not detected; data are mean ± SEM.