Electron microscopy analysis of histone acetylation and DNA strand breaks in mouse elongating spermatids using a dual labelling approach

Nkoma, Geneviève Bikond; Leduc, Frédéric; Jaouad, Leila; Boissonneault, Guylain

doi:10.1111/j.1439-0272.2009.00999.x

Cited by 3 publications

(3 citation statements)

References 7 publications

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“…Accordingly, DNA methyltransferase DNMT1 is regulated by H3K9me3-heterochromatin protein 1α complex and crosstalk between DNMTs and histone methyltransferases is obvious [ 63 – 65 ]. Although histone acetylation in general accompanies gene expression and in some cases causes DNA fragmentation [ 66 ], H3K9me3 is specifically associated with gene silencing and heterochromatin establishment [ 67 , 68 ]. These changes are beneficial for chromatin stability and cell longevity; however, they suppress gene expression [ 67 , 69 , 70 ].…”

Section: Discussionmentioning

confidence: 99%

SIRT1-dependent modulation of methylation and acetylation of histone H3 on lysine 9 (H3K9) in the zygotic pronuclei improves porcine embryo development

Adámková

Petr

et al. 2017

J Animal Sci Biotechnol

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BackgroundThe histone code is an established epigenetic regulator of early embryonic development in mammals. The lysine residue K9 of histone H3 (H3K9) is a prime target of SIRT1, a member of NAD+-dependent histone deacetylase family of enzymes targeting both histone and non-histone substrates. At present, little is known about SIRT1-modulation of H3K9 in zygotic pronuclei and its association with the success of preimplantation embryo development. Therefore, we evaluated the effect of SIRT1 activity on H3K9 methylation and acetylation in porcine zygotes and the significance of H3K9 modifications for early embryonic development.ResultsOur results show that SIRT1 activators resveratrol and BML-278 increased H3K9 methylation and suppressed H3K9 acetylation in both the paternal and maternal pronucleus. Inversely, SIRT1 inhibitors nicotinamide and sirtinol suppressed methylation and increased acetylation of pronuclear H3K9. Evaluation of early embryonic development confirmed positive effect of selective SIRT1 activation on blastocyst formation rate (5.2 ± 2.9% versus 32.9 ± 8.1% in vehicle control and BML-278 group, respectively; P ≤ 0.05). Stimulation of SIRT1 activity coincided with fluorometric signal intensity of ooplasmic ubiquitin ligase MDM2, a known substrate of SIRT1 and known limiting factor of epigenome remodeling.ConclusionsWe conclude that SIRT1 modulates zygotic histone code, obviously through direct deacetylation and via non-histone targets resulting in increased H3K9me3. These changes in zygotes lead to more successful pre-implantation embryonic development and, indeed, the specific SIRT1 activation due to BML-278 is beneficial for in vitro embryo production and blastocyst achievement.Electronic supplementary materialThe online version of this article (10.1186/s40104-017-0214-0) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

SIRT1-dependent modulation of methylation and acetylation of histone H3 on lysine 9 (H3K9) in the zygotic pronuclei improves porcine embryo development

Adámková

Petr

et al. 2017

J Animal Sci Biotechnol

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“…H4, di-acetylated at lysines K5K8 positions, interacts with a single bromodomain (BD1) of double testis-specific, bromodomain (BRDT) which facilitates its displacement [20-22]. Hyperacetylated H4 (H4ac), observed to co-exist with DNA strand breaks in step9-11 elongating murine spermatids, is not co-localised to nicked DNA [23,24]. Rather, the induction of physiological strand breaks in the DNA of step9-11 elongating murine spermatids correlated well with the co-incident expression of topoisomeraseIIβ (TOPIIβ) and tyrosyl DNA phisphodiesterase1 enzymes that induce as well resolve TOPIIβ-mediated DNA nicks as well as γ-H2AX, marker of double strand breaks [25].…”

Section: Introductionmentioning

confidence: 99%

Putative molecular mechanism underlying sperm chromatin remodelling is regulated by reproductive hormones

et al. 2012

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BackgroundThe putative regulatory role of the male reproductive hormones in the molecular mechanism underlying chromatin condensation remains poorly understood. In the past decade, we developed two adult male rat models wherein functional deficits of testosterone or FSH, produced after treatments with 20 mg/Kg/d of cyproterone acetate (CPA) per os, for a period of 15 days or 3 mg/Kg/d of fluphenazine decanoate (FD) subcutaneously, for a period of 60 days, respectively, affected the rate of sperm chromatin decondensation in vitro. These rat models have been used in the current study in order to delineate the putative roles of testosterone and FSH in the molecular mechanism underlying remodelling of sperm chromatin.ResultsWe report that deficits of both testosterone and FSH affected the turnover of polyubiquitylated histones and led to their accumulation in the testis. Functional deficits of testosterone reduced expression of MIWI, the 5-methyl cap binding RNA-binding protein (PIWIlike murine homologue of the Drosophila protein PIWI/P-element induced wimpy testis) containing a PAZ/Piwi-Argonaut-Zwille domain and levels of histone deacetylase1 (HDAC1), ubiquitin ligating enzyme (URE-B1/E3), 20S proteasome α1 concomitant with reduced expression of ubiquitin activating enzyme (ube1), conjugating enzyme (ube2d2), chromodomain Y like protein (cdyl), bromodomain testis specific protein (brdt), hdac6 (histone deacetylase6), androgen-dependent homeobox placentae embryonic protein (pem/RhoX5), histones h2b and th3 (testis-specific h3). Functional deficits of FSH reduced the expression of cdyl and brdt genes in the testis, affected turnover of ubiquitylated histones, stalled the physiological DNA repair mechanism and culminated in spermiation of DNA damaged sperm.ConclusionsWe aver that deficits of both testosterone and FSH differentially affected the process of sperm chromatin remodelling through subtle changes in the ‘chromatin condensation transcriptome and proteome’, thereby stalling the replacement of ‘dynamic’ histones with ‘inert’ protamines, and altering the epigenetic state of condensed sperm chromatin. The inappropriately condensed chromatin affected the sperm chromatin cytoarchitecture, evident from subtle ultrastructural changes in the nuclei of immature caput epididymal sperm of CPA- or FD-treated rats, incubated in vitro with dithiothreitol.

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“…The sperm chromatin state is mainly determined during spermatids’ differentiation (spermiogenesis) resulting from the sequential replacement of the majority of histones by transition proteins and protamines, which are highly basic arginine-rich proteins. Removal of histones is likely facilitated by hyperacetylation [ 5 – 9 ], leading to a more open chromatin state [ 10 ]. From studies using histone deacetylases inhibitors, induced hyperacetylation of histones in cultured cells leads to a greater vulnerability to DNA damage and DNA double-strand breaks (DSBs) are observed [ 11 , 12 ].…”

Section: Introductionmentioning

confidence: 99%

“Breaking news” from spermatids

Gouraud

Brazeau

Grégoire

et al. 2013

Basic Clin. Androl.

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During the haploid phase of spermatogenesis, spermatids undergo a complex remodeling of the paternal genome involving the finely orchestrated replacement of histones by the highly-basic protamines. The associated striking change in DNA topology is characterized by a transient surge of both single- and double-stranded DNA breaks in the whole population of spermatids which are repaired before spermiation. These transient DNA breaks are now considered part of the normal differentiation program of these cells. Despite an increasing interest in the study of spermiogenesis in the last decade and the potential threat to the haploid genome, the origin of these DNA breaks still remains elusive. This review briefly outlines the current hypotheses regarding possible mechanisms that may lead to such transient DNA fragmentation including torsional stress, enzyme-induced breaks, apoptosis-like processes or oxidative stress. A better understanding of the origin of these DNA breaks will lead to further investigations on the genetic instability and mutagenic potential induced by the chromatin remodeling.

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Electron microscopy analysis of histone acetylation and DNA strand breaks in mouse elongating spermatids using a dual labelling approach

Cited by 3 publications

References 7 publications

SIRT1-dependent modulation of methylation and acetylation of histone H3 on lysine 9 (H3K9) in the zygotic pronuclei improves porcine embryo development

SIRT1-dependent modulation of methylation and acetylation of histone H3 on lysine 9 (H3K9) in the zygotic pronuclei improves porcine embryo development

Putative molecular mechanism underlying sperm chromatin remodelling is regulated by reproductive hormones

“Breaking news” from spermatids

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