Effects of the Insemination of Hydrogen Peroxide-Treated Epididymal Mouse Spermatozoa on γH2AX Repair and Embryo Development

Xiao, Jianfeng; Liu, Yanmei; Li, Zhiling; Zhou, Yongcui; Lin, Hong; Wu, Xiaoyan; Chen, Man; Xiao, Wanfen

doi:10.1371/journal.pone.0038742

Cited by 25 publications

(30 citation statements)

References 38 publications

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“…Incubation of sperm with 1500 µM H 2 O 2 did not affect sperm motility, binding or fertilisation, consistent with previous studies [31], [32]. However, as expected we did see increased concentrations of ROS in the sperm and evidence of oxidative damage in the form of increases in sperm mitochondrial ROS and lipid peroxidation.…”

Section: Discussionsupporting

confidence: 92%

Oxidative Stress in Mouse Sperm Impairs Embryo Development, Fetal Growth and Alters Adiposity and Glucose Regulation in Female Offspring

et al. 2014

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Paternal health cues are able to program the health of the next generation however the mechanism for this transmission is unknown. Reactive oxygen species (ROS) are increased in many paternal pathologies, some of which program offspring health, and are known to induce DNA damage and alter the methylation pattern of chromatin. We therefore investigated whether a chemically induced increase of ROS in sperm impairs embryo, pregnancy and offspring health. Mouse sperm was exposed to 1500 µM of hydrogen peroxide (H2O2), which induced oxidative damage, however did not affect sperm motility or the ability to bind and fertilize an oocyte. Sperm treated with H2O2 delayed on-time development of subsequent embryos, decreased the ratio of inner cell mass cells (ICM) in the resulting blastocyst and reduced implantation rates. Crown-rump length at day 18 of gestation was also reduced in offspring produced by H2O2 treated sperm. Female offspring from H2O2 treated sperm were smaller, became glucose intolerant and accumulated increased levels of adipose tissue compared to control female offspring. Interestingly male offspring phenotype was less severe with increases in fat depots only seen at 4 weeks of age, which was restored to that of control offspring later in life, demonstrating sex-specific impacts on offspring. This study implicates elevated sperm ROS concentrations, which are common to many paternal health pathologies, as a mediator of programming offspring for metabolic syndrome and obesity.

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Section: Discussionsupporting

confidence: 92%

Oxidative Stress in Mouse Sperm Impairs Embryo Development, Fetal Growth and Alters Adiposity and Glucose Regulation in Female Offspring

et al. 2014

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“…Despite its importance in DSB repair, H2AX139ph has not been thoroughly investigated during early-embryo development. Few reports have shown the presence of H2AX139ph during embryo development in mice (Luo et al 2006, Adiga et al 2007, Pacchierotti et al 2011, Xiao et al 2012 and rats (Barton et al 2007, Grenier et al 2012, but whether the occurrence of H2AX139ph is correlated with embryo development and quality has not been investigated. Moreover, there were no studies conducted to investigate whether the technologies used for in vitro embryo production affect the occurrence of H2AX139ph.…”

Section: Discussionmentioning

confidence: 99%

Phosphorylated histone H2A.x in porcine embryos produced by IVF and somatic cell nuclear transfer

et al. 2013

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Phosphorylated histone H2A.x (H2AX139ph) is a key factor for the repair of DNA double-strand breaks (DSBs) and the presence of H2AX139ph foci indicates the sites of DSBs. In this study, we characterized the presence of H2AX139ph during in vitro development of porcine embryos produced by IVF and somatic cell nuclear transfer (SCNT). Pronuclear stage embryos produced by IVF had, on average, 9.2 H2AX139ph foci per pronucleus. The number of H2AX139ph foci was higher in the 2-cell-stage embryos than in the 4-cell-stage embryos fixed at 48 h post-fertilization. The percentage of H2AX139ph-positive nuclei was higher in SCNT embryos that were activated with ionomycin (ION) alone than in those activated with ION and strontium chloride (IONCSr 2C ). A negative correlation was found between the percentage of H2AX139ph-positive cells and the total number of cells per embryo in day 7 blastocysts produced by IVF or SCNT. Based on the detection of H2AX139ph foci, the findings of this study indicate that DSBs occur in a high proportion of porcine embryos produced by either IVF or SCNT; fast-cleaving embryos have fewer DSBs than slow-cleaving embryos; the oocyte activation protocol can affect DNA integrity in SCNT embryos; and better-quality blastocysts have fewer DSBs. We propose that the presence of H2AX139ph foci can be a useful marker of embryo quality.

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“…As described in our previous study [21], sperm were collected from the caudae epididymis of mice and incubated in capacitation medium (HTF medium [CooperSurgical, Inc.] containing 1.5% BSA) at 37°C in a 5% CO 2 incubator for 1 h. Female mice were superovulated with consecutive injection of 10 IU pregnant mare serum gonadotropin and 10 IU human chorionic gonadotropin 48 h apart, sacrificed at 13 to 15 h after the human chorionic gonadotropin administration. Fully grown germinal vesicle oocytes were obtained from the ovaries of the mice.…”

Section: Methodsmentioning

confidence: 99%

“…Despite our previous parallel study demonstrated that γH2AX (The DNA damage repair marker) was functional in mouse embryos fertilized with hydrogen peroxide-treated sperm [21], the completeness and correctness of it remain unknown. As stated above, after DNA damage sensing, cell cycle arrest mediated by checkpoint activation would proceed to apoptosis if the damage overwhelmed the repair mechanisms or was incorrectly or partially repaired [15]–[18].…”

Section: Introductionmentioning

confidence: 96%

“…Based on our preestablished DNA-damaged mouse sperm model in which mouse sperm were treated with 1 mM H 2 O 2 to optimally mimic sperm DNA damage caused by cryopreservation-induced ROS, we have discovered that mouse embryos fertilized with hydrogen peroxide-treated sperm showed a delay in cleavage before the blastocyst stage and γH2AX (The DNA damage repair marker) was functional in the early embryos [21]. The delay prompted us the possibility of cell cycle arrest upon checkpoint activation in early mouse embryos similarly fertilized.…”

Section: Introductionmentioning

confidence: 99%

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Zygotic G2/M Cell Cycle Arrest Induced by ATM/Chk1 Activation and DNA Repair in Mouse Embryos Fertilized with Hydrogen Peroxide-Treated Epididymal Mouse Sperm

Wang

et al. 2013

PLoS ONE

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Human sperm cryopreservation for assisted reproduction is compromised by ROS-induced sperm cryodamage. Our previous model study in which mouse sperm were treated with H2O2 to simulate sperm DNA-damage caused by cryopreservation-induced ROS have discovered that mouse embryos fertilized with treated sperm showed a delay in cleavage that might be associated with cell cycle arrest. The DNA-damage checkpoint pathway underlying the delay remained elusive. Moreover, our previous study have also indicated that γH2AX, the DNA-damage repair marker, was functional in mouse embryos similarly fertilized, but the completeness and correctness are unknown and warrant more studies because insufficiency of completeness and correctness of DNA repair would otherwise trigger apoptosis. Based on the aforementioned model, we used embryo culture, inverted microscope, BrdU incorporation and immunofluorescence to explore the cell cycle phase that arrest occurred and the underlying DNA-damage checkpoint pathway in mouse zygotes fertilized with H2O2-treated sperm. We also adopted Tunel to investigate the apoptosis of mouse embryos similarly fertilized at different developmental stages to testify the completeness and correctness of sperm-derived DNA-damage repair. We found G2/M cell cycle arrest in zygotes fertilized with H2O2-treated sperm. ATM (pSer-1981) and Chk1 (pSer-345) activations, rather than ATR (pSer-428) and Chk2 (pThr-68), were detected in zygotes of the treated group. The apoptosis of embryos of different developmental stages of the treated group weren’t different from those of the untreated group. In conclusions, ATM (pSer-1981)-Chk1 (pSer-345) cascade might have mediated G2/M cell cycle arrest and allowed time to facilitate sperm-derived DNA-damage repair in mouse zygotes fertilized with oxygen-stressed sperm, and the DNA-damage repair might be effective.

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Effects of the Insemination of Hydrogen Peroxide-Treated Epididymal Mouse Spermatozoa on γH2AX Repair and Embryo Development

Cited by 25 publications

References 38 publications

Oxidative Stress in Mouse Sperm Impairs Embryo Development, Fetal Growth and Alters Adiposity and Glucose Regulation in Female Offspring

Oxidative Stress in Mouse Sperm Impairs Embryo Development, Fetal Growth and Alters Adiposity and Glucose Regulation in Female Offspring

Phosphorylated histone H2A.x in porcine embryos produced by IVF and somatic cell nuclear transfer

Zygotic G2/M Cell Cycle Arrest Induced by ATM/Chk1 Activation and DNA Repair in Mouse Embryos Fertilized with Hydrogen Peroxide-Treated Epididymal Mouse Sperm

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