Intracellular oxygen metabolism during bovine oocyte and preimplantation embryo development.

Autor: McKeegan PJ; Reproduction and Early Development Research Group, Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, Clarendon Way, Leeds, LS2 9JT, UK. paul.mckeegan@hyms.ac.uk.; Centre for Anatomical and Human Sciences, Hull York Medical School, University of Hull, Hull, HU6 7RX, UK. paul.mckeegan@hyms.ac.uk., Boardman SF; Reproduction and Early Development Research Group, Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, Clarendon Way, Leeds, LS2 9JT, UK.; CARE Fertility, Manchester, England, UK., Wanless AA; Reproduction and Early Development Research Group, Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, Clarendon Way, Leeds, LS2 9JT, UK.; Assisted Conception Unit, Ninewells Hospital, Dundee, Scotland, UK., Boyd G; Reproduction and Early Development Research Group, Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, Clarendon Way, Leeds, LS2 9JT, UK.; Department of Biological Sciences, University of York, Wentworth Way, York, YO10 5DD, England, UK., Warwick LJ; Reproduction and Early Development Research Group, Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, Clarendon Way, Leeds, LS2 9JT, UK.; St James's University Hospital, Beckett Street, Leeds, LS9 7TF, England, UK., Lu J; Reproduction and Early Development Research Group, Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, Clarendon Way, Leeds, LS2 9JT, UK., Gnanaprabha K; Reproduction and Early Development Research Group, Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, Clarendon Way, Leeds, LS2 9JT, UK.; GCRM Fertility, 21 Fifty Pitches Way, Glasgow, G51 4FD, Scotland, UK., Picton HM; Reproduction and Early Development Research Group, Leeds Institute of Cardiovascular and Metabolic Medicine, School of Medicine, University of Leeds, Clarendon Way, Leeds, LS2 9JT, UK.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2021 Oct 28; Vol. 11 (1), pp. 21245. Date of Electronic Publication: 2021 Oct 28.
DOI: 10.1038/s41598-021-99512-5
Abstrakt: We report a novel method to profile intrcellular oxygen concentration (icO 2 ) during in vitro mammalian oocyte and preimplantation embryo development using a commercially available multimodal phosphorescent nanosensor (MM2). Abattoir-derived bovine oocytes and embryos were incubated with MM2 in vitro. A series of inhibitors were applied during live-cell multiphoton imaging to record changes in icO 2 associated with mitochondrial processes. The uncoupler carbonyl cyanide-p-trifluoromethoxyphenylhydrazone (FCCP) uncouples mitochondrial oxygen consumption to its maximum, while antimycin inhibits complex III to ablate mitochondrial oxygen consumption. Increasing oxygen consumption was expected to reduce icO 2 and decreasing oxygen consumption to increase icO 2 . Use of these inhibitors quantifies how much oxygen is consumed at basal in comparison to the upper and lower limits of mitochondrial function. icO 2 measurements were compared to mitochondrial DNA copy number analysed by qPCR. Antimycin treatment increased icO 2 for all stages tested, suggesting significant mitochondrial oxygen consumption at basal. icO 2 of oocytes and preimplantation embryos were unaffected by FCCP treatment. Inner cell mass icO 2 was lower than trophectoderm, perhaps reflecting limitations of diffusion. Mitochondrial DNA copy numbers were similar between stages in the range 0.9-4 × 10 6 copies and did not correlate with icO 2 . These results validate the MM2 probe as a sensitive, non-toxic probe of intracellular oxygen concentration in mammalian oocytes and preimplantation embryos.
(© 2021. The Author(s).)
Databáze: MEDLINE
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