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Oxidative phosphorylation-dependent and -independent oxygen consumption by individual preimplantation mouse embryos

Oxidative phosphorylation-dependent and -independent oxygen consumption by individual preimplantation mouse embryos
Oxidative phosphorylation-dependent and -independent oxygen consumption by individual preimplantation mouse embryos
The self-referencing electrode technique was employed to noninvasively measure gradients of dissolved oxygen in the medium immediately surrounding developing mouse embryos and, thereby, characterized changes in oxygen consumption and utilization during development. A gradient of depleted oxygen surrounded each embryo and could be detected >50 microm from the embryo. Blastocysts depleted the surrounding medium of 0.6+/-0.1 microM of oxygen, whereas early cleavage stage embryos depleted the medium of only 0.3+/-0.1 microM of oxygen, suggesting a twofold increase in oxygen consumption at the blastocyst stage. Mitochondrial oxidative phosphorylation (OXPHOS) accounted for 60-70% of the oxygen consumed by blastocysts, while it accounted for only 30% of the total oxygen consumed by cleavage-stage embryos. The amount of oxygen consumed by non-OXPHOS mechanisms remained relatively constant throughout preimplantation development. By contrast, the amount of oxygen consumed by OXPHOS in blastocysts is greater than that consumed by OXPHOS in cleavage-stage embryos. The amount of oxygen consumed by one-cell embryos was modulated by the absence of pyruvate from the culture medium. Treatment of one-cell embryos and blastocysts with diamide, an agent known to induce cell death in embryos, resulted in a decline in oxygen consumption, such that the medium surrounding dying embryos was not as depleted of oxygen as that surrounding untreated control embryos. Together these results validate the self-referencing electrode technique for analyzing oxygen consumption and utilization by preimplantation embryos and demonstrate that changes in oxygen consumption accompany important physiological events, such as development, response to medium metabolites, or cell death
1866-1874
Trimarchi, James R.
dc15c269-2b07-41fb-b3e5-a9ac457c7994
Liu, Lin
51bc0635-5ce3-4ade-9edf-624ec8a1750b
Porterfield, D. Marshall
ad367ffb-cd69-4c9e-ac0c-f7f048538518
Smith, Peter J.S.
003de469-9420-4f12-8f0e-8e8d76d28d6c
Keefe, David L.
a1d0a08b-d76a-4d64-b2dd-4d1a5fc04451
Trimarchi, James R.
dc15c269-2b07-41fb-b3e5-a9ac457c7994
Liu, Lin
51bc0635-5ce3-4ade-9edf-624ec8a1750b
Porterfield, D. Marshall
ad367ffb-cd69-4c9e-ac0c-f7f048538518
Smith, Peter J.S.
003de469-9420-4f12-8f0e-8e8d76d28d6c
Keefe, David L.
a1d0a08b-d76a-4d64-b2dd-4d1a5fc04451

Trimarchi, James R., Liu, Lin, Porterfield, D. Marshall, Smith, Peter J.S. and Keefe, David L. (2000) Oxidative phosphorylation-dependent and -independent oxygen consumption by individual preimplantation mouse embryos. Biology of Reproduction, 62 (6), 1866-1874. (doi:10.1095/?biolreprod62.6.1866). (PMID:10819794)

Record type: Article

Abstract

The self-referencing electrode technique was employed to noninvasively measure gradients of dissolved oxygen in the medium immediately surrounding developing mouse embryos and, thereby, characterized changes in oxygen consumption and utilization during development. A gradient of depleted oxygen surrounded each embryo and could be detected >50 microm from the embryo. Blastocysts depleted the surrounding medium of 0.6+/-0.1 microM of oxygen, whereas early cleavage stage embryos depleted the medium of only 0.3+/-0.1 microM of oxygen, suggesting a twofold increase in oxygen consumption at the blastocyst stage. Mitochondrial oxidative phosphorylation (OXPHOS) accounted for 60-70% of the oxygen consumed by blastocysts, while it accounted for only 30% of the total oxygen consumed by cleavage-stage embryos. The amount of oxygen consumed by non-OXPHOS mechanisms remained relatively constant throughout preimplantation development. By contrast, the amount of oxygen consumed by OXPHOS in blastocysts is greater than that consumed by OXPHOS in cleavage-stage embryos. The amount of oxygen consumed by one-cell embryos was modulated by the absence of pyruvate from the culture medium. Treatment of one-cell embryos and blastocysts with diamide, an agent known to induce cell death in embryos, resulted in a decline in oxygen consumption, such that the medium surrounding dying embryos was not as depleted of oxygen as that surrounding untreated control embryos. Together these results validate the self-referencing electrode technique for analyzing oxygen consumption and utilization by preimplantation embryos and demonstrate that changes in oxygen consumption accompany important physiological events, such as development, response to medium metabolites, or cell death

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Published date: June 2000
Organisations: University of Southampton

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Local EPrints ID: 190239
URI: http://eprints.soton.ac.uk/id/eprint/190239
PURE UUID: ea76eee1-004c-4f6d-a01f-83d28a7a9bce
ORCID for Peter J.S. Smith: ORCID iD orcid.org/0000-0003-4400-6853

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Date deposited: 20 Jun 2011 09:08
Last modified: 10 Dec 2019 01:38

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Contributors

Author: James R. Trimarchi
Author: Lin Liu
Author: D. Marshall Porterfield
Author: David L. Keefe

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