Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Zinc availability regulates exit from meiosis in maturing mammalian oocytes

Abstract

Cellular metal ion fluxes are known in alkali and alkaline earth metals but are not well documented in transition metals. Here we describe major changes in the zinc physiology of the mammalian oocyte as it matures and initiates embryonic development. Single-cell elemental analysis of mouse oocytes by synchrotron-based X-ray fluorescence microscopy (XFM) revealed a 50% increase in total zinc content within the 12–14-h period of meiotic maturation. Perturbation of zinc homeostasis with a cell-permeable small-molecule chelator blocked meiotic progression past telophase I. Zinc supplementation rescued this phenotype when administered before this meiotic block. However, after telophase arrest, zinc triggered parthenogenesis, suggesting that exit from this meiotic step is tightly regulated by the availability of a zinc-dependent signal. These results implicate the zinc bolus acquired during meiotic maturation as an important part of the maternal legacy to the embryo.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Synchrotron-based X-ray fluorescence microscopy reveals intracellular distribution of the transition elements in oocytes and early embryos.
Figure 2: The heavy-metal chelator TPEN disrupts asymmetric division of the oocyte, which can be rescued by exogenous zinc.
Figure 3: Zinc insufficient oocytes experience a meiotic block following telophase I.
Figure 4: Zinc supplementation following telophase-like arrest induces spontaneous activation of the zinc-insufficient oocyte.
Figure 5: Zinc-insufficient eggs can undergo a true fertilization event but show a delayed pronuclear stage and abnormal calcium oscillations upon activation.
Figure 6: Summary of results.

Similar content being viewed by others

References

  1. Beyersmann, D. & Haase, H. Functions of zinc in signaling, proliferation and differentiation of mammalian cells. Biometals 14, 331–341 (2001).

    Article  CAS  Google Scholar 

  2. Turski, M.L. & Thiele, D.J. New roles for copper metabolism in cell proliferation, signaling, and disease. J. Biol. Chem. 284, 717–721 (2009).

    Article  CAS  Google Scholar 

  3. Zhang, A.S. & Enns, C.A. Iron homeostasis: recently identified proteins provide insight into novel control mechanisms. J. Biol. Chem. 284, 711–715 (2009).

    Article  CAS  Google Scholar 

  4. Yamasaki, S. et al. Zinc is a novel intracellular second messenger. J. Cell Biol. 177, 637–645 (2007).

    Article  CAS  Google Scholar 

  5. Galaris, D., Skiada, V. & Barbouti, A. Redox signaling and cancer: the role of “labile” iron. Cancer Lett. 266, 21–29 (2008).

    Article  CAS  Google Scholar 

  6. Eide, D.J. Zinc transporters and the cellular trafficking of zinc. Biochim. Biophys. Acta 1763, 711–722 (2006).

    Article  CAS  Google Scholar 

  7. Kambe, T., Weaver, B.P. & Andrews, G.K. The genetics of essential metal homeostasis during development. Genesis 46, 214–228 (2008).

    Article  CAS  Google Scholar 

  8. O'Halloran, T.V. Transition metals in control of gene expression. Science 261, 715–725 (1993).

    Article  CAS  Google Scholar 

  9. Valko, M., Morris, H. & Cronin, M.T. Metals, toxicity and oxidative stress. Curr. Med. Chem. 12, 1161–1208 (2005).

    Article  CAS  Google Scholar 

  10. Outten, C.E. & O'Halloran, T.V. Femtomolar sensitivity of metalloregulatory proteins controlling zinc homeostasis. Science 292, 2488–2492 (2001).

    Article  CAS  Google Scholar 

  11. Finney, L.A. & O'Halloran, T.V. Transition metal speciation in the cell: insights from the chemistry of metal ion receptors. Science 300, 931–936 (2003).

    Article  CAS  Google Scholar 

  12. Bruinsma, J.J., Jirakulaporn, T., Muslin, A.J. & Kornfeld, K. Zinc ions and cation diffusion facilitator proteins regulate Ras-mediated signaling. Dev. Cell 2, 567–578 (2002).

    Article  CAS  Google Scholar 

  13. Nomizu, T., Falchuk, K.H. & Vallee, B.L. Zinc, iron, and copper contents of Xenopus laevis oocytes and embryos. Mol. Reprod. Dev. 36, 419–423 (1993).

    Article  CAS  Google Scholar 

  14. Sun, L., Chai, Y., Hannigan, R., Bhogaraju, V.K. & Machaca, K. Zinc regulates the ability of Cdc25C to activate MPF/cdk1. J. Cell. Physiol. 213, 98–104 (2007).

    Article  CAS  Google Scholar 

  15. Falchuk, K.H. & Montorzi, M. Zinc physiology and biochemistry in oocytes and embryos. Biometals 14, 385–395 (2001).

    Article  CAS  Google Scholar 

  16. Falchuk, K.H., Montorzi, M. & Vallee, B.L. Zinc uptake and distribution in Xenopus laevis oocytes and embryos. Biochemistry 34, 16524–16531 (1995).

    Article  CAS  Google Scholar 

  17. Stitzel, M.L. & Seydoux, G. Regulation of the oocyte-to-zygote transition. Science 316, 407–408 (2007).

    Article  CAS  Google Scholar 

  18. Gosden, R.G. Oogenesis as a foundation for embryogenesis. Mol. Cell. Endocrinol. 186, 149–153 (2002).

    Article  CAS  Google Scholar 

  19. Gandolfi, T.A. & Gandolfi, F. The maternal legacy to the embryo: cytoplasmic components and their effects on early development. Theriogenology 55, 1255–1276 (2001).

    Article  CAS  Google Scholar 

  20. Jeruss, J.S. & Woodruff, T.K. Preservation of fertility in patients with cancer. N. Engl. J. Med. 360, 902–911 (2009).

    Article  CAS  Google Scholar 

  21. Picton, H., Briggs, D. & Gosden, R. The molecular basis of oocyte growth and development. Mol. Cell. Endocrinol. 145, 27–37 (1998).

    Article  CAS  Google Scholar 

  22. Perreault, S.D., Barbee, R.R. & Slott, V.L. Importance of glutathione in the acquisition and maintenance of sperm nuclear decondensing activity in maturing hamster oocytes. Dev. Biol. 125, 181–186 (1988).

    Article  CAS  Google Scholar 

  23. Ajduk, A., Malagocki, A. & Maleszewski, M. Cytoplasmic maturation of mammalian oocytes: development of a mechanism responsible for sperm-induced Ca2+ oscillations. Reprod. Biol. 8, 3–22 (2008).

    Article  Google Scholar 

  24. Cooley, L. Oogenesis: variations on a theme. Dev. Genet. 16, 1–5 (1995).

    Article  CAS  Google Scholar 

  25. Martell, A.E. & Smith, R.M. NIST critical stability constants of metal complexes. in NIST Standard Reference Database 46, v5.0 (Plenum, New York, 1998).

  26. Suhy, D.A., Simon, K.D., Linzer, D.I. & O'Halloran, T.V. Metallothionein is part of a zinc-scavenging mechanism for cell survival under conditions of extreme zinc deprivation. J. Biol. Chem. 274, 9183–9192 (1999).

    Article  CAS  Google Scholar 

  27. Arslan, P., Di Virgilio, F., Beltrame, M., Tsien, R.Y. & Pozzan, T. Cytosolic Ca2+ homeostasis in Ehrlich and Yoshida carcinomas. A new, membrane-permeant chelator of heavy metals reveals that these ascites tumor cell lines have normal cytosolic free Ca2+. J. Biol. Chem. 260, 2719–2727 (1985).

    CAS  PubMed  Google Scholar 

  28. Barrett, S.L. & Albertini, D.F. Allocation of gamma-tubulin between oocyte cortex and meiotic spindle influences asymmetric cytokinesis in the mouse oocyte. Biol. Reprod. 76, 949–957 (2007).

    Article  CAS  Google Scholar 

  29. Brunet, S. & Maro, B. Cytoskeleton and cell cycle control during meiotic maturation of the mouse oocyte: integrating time and space. Reproduction 130, 801–811 (2005).

    Article  CAS  Google Scholar 

  30. Santos, F. & Dean, W. Using immunofluorescence to observe methylation changes in mammalian preimplantation embryos. Methods Mol. Biol. 325, 129–137 (2006).

    CAS  PubMed  Google Scholar 

  31. Santos, F., Hendrich, B., Reik, W. & Dean, W. Dynamic reprogramming of DNA methylation in the early mouse embryo. Dev. Biol. 241, 172–182 (2002).

    Article  CAS  Google Scholar 

  32. Stricker, S.A. Comparative biology of calcium signaling during fertilization and egg activation in animals. Dev. Biol. 211, 157–176 (1999).

    Article  CAS  Google Scholar 

  33. Ducibella, T. et al. Egg-to-embryo transition is driven by differential responses to Ca2+ oscillation number. Dev. Biol. 250, 280–291 (2002).

    Article  CAS  Google Scholar 

  34. Tóth, S., Huneau, D., Banrezes, B. & Ozil, J.P. Egg activation is the result of calcium signal summation in the mouse. Reproduction 131, 27–34 (2006).

    Article  Google Scholar 

  35. Nagy, A., Gertsenstein, M., Vintersten, K. & Behringer, R. (eds.). Manipulating the Mouse Embryo: A Laboratory Manual (Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, USA, 2003).

  36. Taki, M., Wolford, J.L. & O'Halloran, T.V. Emission ratiometric imaging of intracellular zinc: design of a benzoxazole fluorescent sensor and its application in two-photon microscopy. J. Am. Chem. Soc. 126, 712–713 (2004).

    Article  CAS  Google Scholar 

  37. Wickramasinghe, D., Ebert, K.M. & Albertini, D.F. Meiotic competence acquisition is associated with the appearance of M-phase characteristics in growing mouse oocytes. Dev. Biol. 143, 162–172 (1991).

    Article  CAS  Google Scholar 

  38. Sorensen, R.A. & Wassarman, P.M. Relationship between growth and meiotic maturation of the mouse oocyte. Dev. Biol. 50, 531–536 (1976).

    Article  CAS  Google Scholar 

  39. Erickson, G.F. & Sorensen, R.A. In vitro maturation of mouse oocytes isolated from late, middle, and pre-antral graafian follicles. J. Exp. Zool. 190, 123–127 (1974).

    Article  CAS  Google Scholar 

  40. Araki, K. et al. Meiotic abnormalities of c-mos knockout mouse oocytes: activation after first meiosis or entrance into third meiotic metaphase. Biol. Reprod. 55, 1315–1324 (1996).

    Article  CAS  Google Scholar 

  41. Eppig, J.J., Schultz, R.M., O'Brien, M. & Chesnel, F. Relationship between the developmental programs controlling nuclear and cytoplasmic maturation of mouse oocytes. Dev. Biol. 164, 1–9 (1994).

    Article  CAS  Google Scholar 

  42. Ozil, J.P., Banrezes, B., Toth, S., Pan, H. & Schultz, R.M. Ca2+ oscillatory pattern in fertilized mouse eggs affects gene expression and development to term. Dev. Biol. 300, 534–544 (2006).

    Article  CAS  Google Scholar 

  43. Vogt, S. MAPS: A set of software tools for analysis and visualization of 3D X-ray fluorescence data sets. J. Phys. IV France 104, 635–638 (2003).

    Article  CAS  Google Scholar 

  44. Ibáñez, E., Sanfins, A., Combelles, C.M., Overstrom, E.W. & Albertini, D.F. Genetic strain variations in the metaphase-II phenotype of mouse oocytes matured in vivo or in vitro. Reproduction 130, 845–855 (2005).

    Article  Google Scholar 

Download references

Acknowledgements

The authors gratefully acknowledge J. Jozefik, S. Kiesewetter and D. Mackovic for animal care and concerns. We would also like to thank the P01 Histology Core (T. Wellington, director), the Analytical Services Laboratory and the Quantitative Bioelement Imaging Center in the Chemistry of Life Processes Institute at Northwestern University for reagents and discussions regarding sample processing. This work is supported by US National Institutes of Health grants P01 HD021921 and GM38784, the W.M. Keck Foundation Medical Research Award and the Chicago Biomedical Consortium Spark Award. A.M.K. was a fellow of the Reproductive Biology Training Grant HD007068. Use of the Advanced Photon Source at Argonne National Laboratory was supported by the Office of Basic Energy Sciences in the Office of Science of the US Department of Energy, under contract no. DE-AC02-06CH11357.

Author information

Authors and Affiliations

Authors

Contributions

A.M.K., T.V.O. and T.K.W. designed the research and wrote the manuscript. A.M.K. performed the research. S.V. provided XFM data analysis and technical support.

Corresponding authors

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

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Methods, Supplementary Scheme 1, Supplementary Figures 1–4 and Supplementary Tables 1–3 (PDF 9941 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kim, A., Vogt, S., O'Halloran, T. et al. Zinc availability regulates exit from meiosis in maturing mammalian oocytes. Nat Chem Biol 6, 674–681 (2010). https://doi.org/10.1038/nchembio.419

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nchembio.419

This article is cited by

Search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing