Readthrough acetylcholinesterase (AChE-R) and regulated necrosis

pharmacological targets for the regulation of ovarian functions?

J. Blohberger, L. Kunz, D. Einwang, U. Berg, D. Berg, Sergio Ojeda, Gregory Dissen, T. Fröhlich, G. J. Arnold, H. Soreq, H. Lara, A. Mayerhofer

    Research output: Contribution to journalArticle

    21 Citations (Scopus)

    Abstract

    Proliferation, differentiation and death of ovarian cells ensure orderly functioning of the female gonad during the reproductive phase, which ultimately ends with menopause in women. These processes are regulated by several mechanisms, including local signaling via neurotransmitters. Previous studies showed that ovarian non-neuronal endocrine cells produce acetylcholine (ACh), which likely acts as a trophic factor within the ovarian follicle and the corpus luteum via muscarinic ACh receptors. How its actions are restricted was unknown. We identified enzymatically active acetylcholinesterase (AChE) in human ovarian follicular fluid as a product of human granulosa cells. AChE breaks down ACh and thereby attenuates its trophic functions. Blockage of AChE by huperzine A increased the trophic actions as seen in granulosa cells studies. Among ovarian AChE variants, the readthrough isoform AChE-R was identified, which has further, non-enzymatic roles. AChE-R was found in follicular fluid, granulosa and theca cells, as well as luteal cells, implying that such functions occur in vivo. A synthetic AChE-R peptide (ARP) was used to explore such actions and induced in primary, cultured human granulosa cells a caspase-independent form of cell death with a distinct balloon-like morphology and the release of lactate dehydrogenase. The RIPK1 inhibitor necrostatin-1 and the MLKL-blocker necrosulfonamide significantly reduced this form of cell death. Thus a novel non-enzymatic function of AChE-R is to stimulate RIPK1/MLKL-dependent regulated necrosis (necroptosis). The latter complements a cholinergic system in the ovary, which determines life and death of ovarian cells. Necroptosis likely occurs in the primate ovary, as granulosa and luteal cells were immunopositive for phospho-MLKL, and hence necroptosis may contribute to follicular atresia and luteolysis. The results suggest that interference with the enzymatic activities of AChE and/or interference with necroptosis may be novel approaches to influence ovarian functions.

    Original languageEnglish (US)
    Pages (from-to)e1685
    JournalCell Death and Disease
    Volume6
    DOIs
    StatePublished - 2015

    Fingerprint

    Acetylcholinesterase
    Pharmacology
    Granulosa Cells
    Cell Death
    Luteal Cells
    Follicular Fluid
    Acetylcholine
    Ovary
    Follicular Atresia
    Theca Cells
    Luteolysis
    Ovarian Follicle
    Endocrine Cells
    Corpus Luteum
    Gonads
    Muscarinic Receptors
    Menopause
    Caspases
    L-Lactate Dehydrogenase
    Cholinergic Agents

    ASJC Scopus subject areas

    • Medicine(all)

    Cite this

    Readthrough acetylcholinesterase (AChE-R) and regulated necrosis : pharmacological targets for the regulation of ovarian functions? / Blohberger, J.; Kunz, L.; Einwang, D.; Berg, U.; Berg, D.; Ojeda, Sergio; Dissen, Gregory; Fröhlich, T.; Arnold, G. J.; Soreq, H.; Lara, H.; Mayerhofer, A.

    In: Cell Death and Disease, Vol. 6, 2015, p. e1685.

    Research output: Contribution to journalArticle

    Blohberger, J, Kunz, L, Einwang, D, Berg, U, Berg, D, Ojeda, S, Dissen, G, Fröhlich, T, Arnold, GJ, Soreq, H, Lara, H & Mayerhofer, A 2015, 'Readthrough acetylcholinesterase (AChE-R) and regulated necrosis: pharmacological targets for the regulation of ovarian functions?', Cell Death and Disease, vol. 6, pp. e1685. https://doi.org/10.1038/cddis.2015.51
    Blohberger, J. ; Kunz, L. ; Einwang, D. ; Berg, U. ; Berg, D. ; Ojeda, Sergio ; Dissen, Gregory ; Fröhlich, T. ; Arnold, G. J. ; Soreq, H. ; Lara, H. ; Mayerhofer, A. / Readthrough acetylcholinesterase (AChE-R) and regulated necrosis : pharmacological targets for the regulation of ovarian functions?. In: Cell Death and Disease. 2015 ; Vol. 6. pp. e1685.
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    abstract = "Proliferation, differentiation and death of ovarian cells ensure orderly functioning of the female gonad during the reproductive phase, which ultimately ends with menopause in women. These processes are regulated by several mechanisms, including local signaling via neurotransmitters. Previous studies showed that ovarian non-neuronal endocrine cells produce acetylcholine (ACh), which likely acts as a trophic factor within the ovarian follicle and the corpus luteum via muscarinic ACh receptors. How its actions are restricted was unknown. We identified enzymatically active acetylcholinesterase (AChE) in human ovarian follicular fluid as a product of human granulosa cells. AChE breaks down ACh and thereby attenuates its trophic functions. Blockage of AChE by huperzine A increased the trophic actions as seen in granulosa cells studies. Among ovarian AChE variants, the readthrough isoform AChE-R was identified, which has further, non-enzymatic roles. AChE-R was found in follicular fluid, granulosa and theca cells, as well as luteal cells, implying that such functions occur in vivo. A synthetic AChE-R peptide (ARP) was used to explore such actions and induced in primary, cultured human granulosa cells a caspase-independent form of cell death with a distinct balloon-like morphology and the release of lactate dehydrogenase. The RIPK1 inhibitor necrostatin-1 and the MLKL-blocker necrosulfonamide significantly reduced this form of cell death. Thus a novel non-enzymatic function of AChE-R is to stimulate RIPK1/MLKL-dependent regulated necrosis (necroptosis). The latter complements a cholinergic system in the ovary, which determines life and death of ovarian cells. Necroptosis likely occurs in the primate ovary, as granulosa and luteal cells were immunopositive for phospho-MLKL, and hence necroptosis may contribute to follicular atresia and luteolysis. The results suggest that interference with the enzymatic activities of AChE and/or interference with necroptosis may be novel approaches to influence ovarian functions.",
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    AU - Ojeda, Sergio

    AU - Dissen, Gregory

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