Mobilisation of Ca2+ stores and flagellar regulation in human sperm by S-nitrosylation: a role for NO synthesised in the female reproductive tract

Gisela Machado-Oliveira, Linda Lefievre, Christopher Ford, M. Belen Herrero, Christopher Barratt, Thomas J. Connolly, Katherine Nash, Aduen Morales-Garcia, Jackson Kirkman-Brown, Steve Publicover

    Research output: Contribution to journalArticle

    28 Citations (Scopus)

    Abstract

    Generation of NO by nitric oxide synthase (NOS) is implicated in gamete interaction and fertilisation. Exposure of human spermatozoa to NO donors caused mobilisation of stored Ca2+ by a mechanism that did not require activation of guanylate cyclase but was mimicked by S-nitroso-glutathione (GSNO; an S-nitrosylating agent). Application of dithiothreitol, to reduce protein -SNO groups, rapidly reversed the actions of NO and GSNO on [Ca2+](i). The effects of NO, GSNO and dithiothreitol on sperm protein S-nitrosylation, assessed using the biotin switch method, closely paralleled their actions on [Ca2+](i). Immunofluorescent staining revealed constitutive and inducible NOS in human oviduct and cumulus (the cellular layer investing the oocyte). 4,5-diaminofluorescein (DAF) staining demonstrated production of NO by these tissues. Incubation of human sperm with oviduct explants induced sperm protein S-nitrosylation resembling that induced by NO donors and GSNO. Progesterone (a product of cumulus cells) also mobilises stored Ca2+ in human sperm. Pre-treatment of sperm with NO greatly enhanced the effect of progesterone on [Ca2+](i), resulting in a prolonged increase in flagellar excursion. We conclude that NO regulates mobilisation of stored Ca2+ in human sperm by protein S-nitrosylation, that this action is synergistic with that of progesterone and that this synergism is potentially highly significant in gamete interactions leading to fertilisation.

    Original languageEnglish
    Pages (from-to)3677-3686
    Number of pages10
    JournalDevelopment
    Volume135
    Issue number22
    Early online date8 Oct 2008
    DOIs
    Publication statusPublished - 2008

    Keywords

    • Calcium
    • Cumulus
    • Motility
    • Nitric oxide
    • Oviduct
    • Sperm
    • Human
    • Nitric oxide synthase
    • Mitochondrial permeability transition
    • Adenine nucleotide translocase
    • Sensitive guanylyl cyclase
    • Human spermatozoa
    • Acrosome reaction
    • In vitro
    • Cumulus cells
    • L-arginine
    • Embryonic development

    Research Output

    • 28 Citations
    • 1 Article

    Communication between female tract and sperm: saying NO* when you mean yes

    Lefièvre, L., Machado-Oliveira, G., Ford, C., Kirkman-Brown, J., Barratt, C. & Publicover, S., 2009, In : Communicative & Integrative Biology. 2, 2, p. 82-5 4 p.

    Research output: Contribution to journalArticle

  • Cite this

    Machado-Oliveira, G., Lefievre, L., Ford, C., Herrero, M. B., Barratt, C., Connolly, T. J., Nash, K., Morales-Garcia, A., Kirkman-Brown, J., & Publicover, S. (2008). Mobilisation of Ca2+ stores and flagellar regulation in human sperm by S-nitrosylation: a role for NO synthesised in the female reproductive tract. Development, 135(22), 3677-3686. https://doi.org/10.1242/dev.024521