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

    26 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

    Cite this

    Machado-Oliveira, Gisela ; Lefievre, Linda ; Ford, Christopher ; Herrero, M. Belen ; Barratt, Christopher ; Connolly, Thomas J. ; Nash, Katherine ; Morales-Garcia, Aduen ; Kirkman-Brown, Jackson ; Publicover, Steve. / Mobilisation of Ca2+ stores and flagellar regulation in human sperm by S-nitrosylation : a role for NO synthesised in the female reproductive tract. In: Development. 2008 ; Vol. 135, No. 22. pp. 3677-3686.
    @article{22b25a619f5347d1aa38dc27f80248c9,
    title = "Mobilisation of Ca2+ stores and flagellar regulation in human sperm by S-nitrosylation: a role for NO synthesised in the female reproductive tract",
    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.",
    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",
    author = "Gisela Machado-Oliveira and Linda Lefievre and Christopher Ford and Herrero, {M. Belen} and Christopher Barratt and Connolly, {Thomas J.} and Katherine Nash and Aduen Morales-Garcia and Jackson Kirkman-Brown and Steve Publicover",
    year = "2008",
    doi = "10.1242/dev.024521",
    language = "English",
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    Machado-Oliveira, G, Lefievre, L, Ford, C, Herrero, MB, Barratt, C, Connolly, TJ, 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, vol. 135, no. 22, pp. 3677-3686. https://doi.org/10.1242/dev.024521

    Mobilisation of Ca2+ stores and flagellar regulation in human sperm by S-nitrosylation : a role for NO synthesised in the female reproductive tract. / Machado-Oliveira, Gisela; Lefievre, Linda; Ford, Christopher; Herrero, M. Belen; Barratt, Christopher; Connolly, Thomas J.; Nash, Katherine; Morales-Garcia, Aduen; Kirkman-Brown, Jackson; Publicover, Steve.

    In: Development, Vol. 135, No. 22, 2008, p. 3677-3686.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Mobilisation of Ca2+ stores and flagellar regulation in human sperm by S-nitrosylation

    T2 - a role for NO synthesised in the female reproductive tract

    AU - Machado-Oliveira, Gisela

    AU - Lefievre, Linda

    AU - Ford, Christopher

    AU - Herrero, M. Belen

    AU - Barratt, Christopher

    AU - Connolly, Thomas J.

    AU - Nash, Katherine

    AU - Morales-Garcia, Aduen

    AU - Kirkman-Brown, Jackson

    AU - Publicover, Steve

    PY - 2008

    Y1 - 2008

    N2 - 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.

    AB - 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.

    KW - Calcium

    KW - Cumulus

    KW - Motility

    KW - Nitric oxide

    KW - Oviduct

    KW - Sperm

    KW - Human

    KW - Nitric oxide synthase

    KW - Mitochondrial permeability transition

    KW - Adenine nucleotide translocase

    KW - Sensitive guanylyl cyclase

    KW - Human spermatozoa

    KW - Acrosome reaction

    KW - In vitro

    KW - Cumulus cells

    KW - L-arginine

    KW - Embryonic development

    U2 - 10.1242/dev.024521

    DO - 10.1242/dev.024521

    M3 - Article

    VL - 135

    SP - 3677

    EP - 3686

    JO - Development

    JF - Development

    SN - 0950-1991

    IS - 22

    ER -