Non-invasive imaging of mouse embryo metabolism in response to induced hypoxia
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EMBRYO BIOLOGY
Non-invasive imaging of mouse embryo metabolism in response to induced hypoxia Emily A. Seidler 1,2,3
&
T. Sanchez 1 & M. Venturas 1 & D. Sakkas 3 & D. J. Needleman 1
Received: 21 April 2020 / Accepted: 25 June 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract Purpose This study used noninvasive, fluorescence lifetime imaging microscopy (FLIM)-based imaging of NADH and FAD to characterize the metabolic response of mouse embryos to short-term oxygen deprivation. We investigated the response to hypoxia at various preimplantation stages. Methods Mouse oocytes and embryos were exposed to transient hypoxia by dropping the oxygen concentration in media from 5–0% over the course of ~1.5 h, then 5% O2 was restored. During this time, FLIM-based metabolic imaging measurements of oocyte/embryo cohorts were taken every 3 minutes. Experiments were performed in triplicate for oocytes and embryos at the 1- to 8-cell, morula, and blastocyst stages. Maximum hypoxia response for each of eight measured quantitative FLIM parameters was taken from the time points immediately before oxygen restoration. Results Metabolic profiles showed significant changes in response to hypoxia for all stages of embryo development. The response of the eight measured FLIM parameters to hypoxia was highly stage-dependent. Of the eight FLIM parameters measured, NADH and FAD intensity showed the most dramatic metabolic responses in early developmental stages. At later stages, however, other parameters, such as NADH fraction engaged and FAD lifetimes, showed greater changes. Metabolic parameter values generally returned to baseline with the restoration of 5% oxygen. Conclusions Quantitative FLIM-based metabolic imaging was highly sensitive to metabolic changes induced by hypoxia. Metabolic response profiles to oxygen deprivation were distinct at different stages, reflecting differences in metabolic plasticity as preimplantation embryos develop. Keywords Embryo metabolism . Hypoxia . Noninvasive assessment . Metabolic imaging . FLIM . Mitochondria
Introduction
D. Sakkas and D. J. Needleman are the co-senior authors Electronic supplementary material The online version of this article (https://doi.org/10.1007/s10815-020-01872-w) contains supplementary material, which is available to authorized users. * Emily A. Seidler [email protected] 1
Department of Molecular and Cellular Biology and John A. Paulson School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts, USA
2
Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA, USA
3
Boston IVF, Waltham, MA, USA
In the 1960s, it was established that metabolites such as glucose, pyruvate, and lactate are essential for mammalian embryo development in vitro [1–4]. Further studies showed that there is an intricate relationship between specific metabolites and embryo metabolism over the course of an embryo’s preimplantation development [5–8]. For example, early in development (pre-compaction), embryos rely
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