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NO is necessary and sufficient for egg activation at fertilization

Nature volume 406, pages 633–636 (2000)Cite this article

Abstract

The early steps that lead to the rise in calcium and egg activation at fertilization are unknown but of great interest—particularly with the advent of in vitro fertilization techniques for treating male infertility and whole-animal cloning by nuclear transfer. This calcium rise is required for egg activation and the subsequent events of development in eggs of all species1,2. Injection of intact sperm or sperm extracts can activate eggs, suggesting that sperm-derived factors may be involved. Here we show that nitric oxide synthase is present at high concentration and active in sperm after activation by the acrosome reaction. An increase in nitrosation within eggs is evident seconds after insemination and precedes the calcium pulse of fertilization. Microinjection of nitric oxide donors or recombinant nitric oxide synthase recapitulates events of egg activation, whereas prior injection of oxyhaemoglobin, a physiological nitric oxide scavenger, prevents egg activation after fertilization. We conclude that nitric oxide synthase and nitric-oxide-related bioactivity satisfy the primary criteria of an egg activator: they are present in an appropriate place, active at an appropriate time, and are necessary and sufficient for successful fertilization.

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Figure 1: NOS in S. purpuratus gametes.
Figure 2: NOS activity in S. purpuratus gametes.
Figure 3: Nitrosation after S. purpuratus gamete activation.
Figure 4: NO is necessary and sufficient for egg activation.

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Acknowledgements

We thank H. Tosuji for assistance with DAF, V. Lance for haemoglobin preparation, R. Lewis and H. Schulman for helpful comments, and K. Foltz and S. Feinstein for their support during R.C.K.'s stay at UCSB. RCK is a pre-doctoral fellow of the Howard Hughes Medical Institute. This work was supported by NIH grants to D.E. and J.B. and NSF grants to S.S. and S.T.

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Authors and Affiliations

  1. Neurosciences Program, Stanford University School of Medicine, Stanford University, Stanford, 94305, California, USA

    Richard C. Kuo & Stuart H. Thompson

  2. Department of Biological Sciences Hopkins Marine Station, Stanford University, Pacific Grove, 93950, California, USA

    Richard C. Kuo, Stuart H. Thompson, Chris Patton & David Epel

  3. Cornell Nanofabrication Facility, Knight Laboratory, Cornell University, Ithaca, 14853, New York , USA

    Gregory T. Baxter

  4. Department of Biology, University of New Mexico, Albuquerque, 87131 , New Mexico, USA

    Stephen A. Stricker

  5. Department of Cell Biology, Duke University Medical Center, Durham, 27710, North Carolina, USA

    Joseph Bonaventura

  6. Nicholas School of the Environment, Duke Marine Biomedical Center, Pivers Island, 28516, North Carolina , USA

    Joseph Bonaventura

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  1. Richard C. Kuo
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Correspondence to David Epel.

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Kuo, R., Baxter, G., Thompson, S. et al. NO is necessary and sufficient for egg activation at fertilization. Nature 406, 633–636 (2000). https://doi.org/10.1038/35020577

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