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Quantitative mapping of zinc fluxes in the mammalian egg reveals the origin of fertilization-induced zinc sparks

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Abstract

Fertilization of a mammalian egg initiates a series of ‘zinc sparks’ that are necessary to induce the egg-to-embryo transition. Despite the importance of these zinc-efflux events little is known about their origin. To understand the molecular mechanism of the zinc spark we combined four physical approaches that resolve zinc distributions in single cells: a chemical probe for dynamic live-cell fluorescence imaging and a combination of scanning transmission electron microscopy with energy-dispersive spectroscopy, X-ray fluorescence microscopy and three-dimensional elemental tomography for high-resolution elemental mapping. We show that the zinc spark arises from a system of thousands of zinc-loaded vesicles, each of which contains, on average, 106 zinc atoms. These vesicles undergo dynamic movement during oocyte maturation and exocytosis at the time of fertilization. The discovery of these vesicles and the demonstration that zinc sparks originate from them provides a quantitative framework for understanding how zinc fluxes regulate cellular processes.

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Figure 1: Vital zinc probe reveals cortical compartments in the female gamete in mouse.
Figure 2: Labile zinc is cortically localized in the oocyte and tracks with CG staining.
Figure 3: Zinc fixation enables ultrastructural identification of zinc-enriched cortical compartments by STEM-EDS.
Figure 4: XFM and tomography provide zinc quantification and mapping within the egg.
Figure 5: Live-cell fluorescence zinc imaging demonstrates that intracellular zinc compartments are the source of the extracellular zinc spark.
Figure 6: The zinc flux during egg activation is regulated by a quantitative loss of cortical zinc compartments.

Change history

  • 23 December 2014

    In the version of this Article previously published online, the NIH grant number T32GM105538 was missing from the Acknowledgements section. This has now been corrected in all versions of the Article.

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Acknowledgements

The authors thank members of the O'Halloran, Woodruff and Dravid labs for scientific discussions and advice. We thank E. W. Roth for the preparation of electron microscopy samples and J-H. Chung and J. Shangguan for help with chemical syntheses. Equipment and experimental guidance were provided by the following core facilities at Northwestern University: the Integrated Molecular Structure Education and Research Center, the Biological Imaging Facility, the Quantitative Bioelemental Imaging Center, the Electron Probe Instrumentation Centre and the Keck Biophysics Facility. This research used resources of the Advanced Photon Source, a US Department of Energy (DOE) Office of Science User Facility operated for the DOE Office of Science by Argonne National Laboratory under contract no. DE-AC02-06CH11357. This work was supported by a Medical Research Award from the W. M. Keck Foundation, a SPARK Award from the Chicago Biomedical Consortium and the National Institutes of Health (P01 HD021921, GM38784, U54HD076188 and T32GM105538).

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E.L.Q., R.B., F.E.D., B.Y.K., V.P.D., T.K.W. and T.V.O. designed the research. E.L.Q., R.B., F.E.D., B.Y.K., S.A.G. and A.R.B. performed the research. S.C.G., S.V. and S.C. helped design and implement XFM experiments and process and analyse the data. E.L.Q., R.B., F.E.D., T.K.W. and T.V.O. wrote the manuscript. All the authors discussed the results and commented on the manuscript.

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Correspondence to Teresa K. Woodruff or Thomas V. O'Halloran.

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Que, E., Bleher, R., Duncan, F. et al. Quantitative mapping of zinc fluxes in the mammalian egg reveals the origin of fertilization-induced zinc sparks. Nature Chem 7, 130–139 (2015). https://doi.org/10.1038/nchem.2133

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