Phospho-Cdc25 correlates with activating G2/M checkpoint in mouse zygotes fertilized with hydrogen peroxide-treated mouse sperm
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The presence of oxidative stress in sperm cryopreservation induces sperm DNA damage. Our previous study has discovered that γH2AX, the DNA-damaged marker, was activated in the early mouse embryos fertilized with hydrogen peroxide (H2O2)-treated sperm. Furthermore, we found that checkpoint proteins ATM and Chk1 were phosphorylated and activated in the early mouse embryos. On the basis of previous researches, we examined the effects of sperm DNA damage on cell cycle arrest in mouse zygotes fertilized with H2O2-treated sperm. Development of fertilized eggs arrested at the PN disappearance stage. At 19 and 24 hours post-insemination (hpi), the percentage of zygotes at the PN disappearance stage was higher in H2O2-treated group compared to the control group. Immunofluorescence staining revealed Phospho-Cdc25C (Ser216) and Phospho-Cdc25B (Ser323) in or surrounding a single pronucleus, following insemination with H2O2-treated sperm. Our study suggests that fertilization with DNA-damaged sperm results in cell cycle arrest mediated by G2/M checkpoint activation in one of the pronuclei in mouse zygotes fertilized with H2O2-treated sperm; Phospho-Cdc25C and Phospho-Cdc25B correlate with activating G2/M checkpoint in zygotes fertilized with H2O2-treated sperm.
KeywordsSperm cryopreservation Oxidative stress Embryo development G2/M checkpoint Cdc25
We are grateful for the support from Center for Neuroscience, Shantou University Medical College for the utilization of the laser confocal microscopy.
Conflict of interest
The authors have declared that no conflict of interest exists.
This study was supported by the National Natural Science Foundation of China (No. 81070542 and No. 30872771), the Natural Science Foundation of Guangdong Province of China (No. 10151503102000020 and No. 81515031102000010). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
- 3.Lim JJ, Shin TE, Song SH, Bak CW, Yoon TK, Lee DR (2010) Effect of liquid nitrogen vapor storage on the motility, viability, morphology, deoxyribonucleic acid integrity, and mitochondrial potential of frozen–thawed human spermatozoa. Fertil Steril 94:2736–2741. doi: 10.1016/j.fertnstert.2010.02.051 PubMedCrossRefGoogle Scholar
- 21.Wang B, Li Z, Wang C, Chen M, Xiao J, Wu X, Xiao W, Song Y, Wang X (2013) 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. PLoS ONE 8:e73987. doi: 10.1371/journal.pone.0073987 PubMedCrossRefPubMedCentralGoogle Scholar
- 28.Virro MR, Larson-Cook KL, Evenson DP (2004) Sperm chromatin structure assay (SCSA) parameters are related to fertilization, blastocyst development, and ongoing pregnancy in in vitro fertilization and intracytoplasmic sperm injection cycles. Fertil Steril 81:1289–1295. doi: 10.1016/j.fertnstert.2003.09.063 PubMedCrossRefGoogle Scholar
- 29.Pregl Breznik B, Kovacic B, Vlaisavljevic V (2013) Are sperm DNA fragmentation, hyperactivation, and hyaluronan-binding ability predictive for fertilization and embryo development in in vitro fertilization and intracytoplasmic sperm injection? Fertil Steril 99:1233–1241. doi: 10.1016/j.fertnstert.2012.11.048 PubMedCrossRefGoogle Scholar
- 31.Upadhya D, Kalthur G, Kumar P, Rao BS, Adiga SK (2010) Association between the extent of DNA damage in the spermatozoa, fertilization and developmental competence in preimplantation stage embryos. J Turk Ger Gynecol Assoc 11:182–186. doi: 10.5152/jtgga.2010.34 PubMedCrossRefPubMedCentralGoogle Scholar
- 38.Chen F, He ML, Bower J, Sbarra D, C M Lin M, Kung HF and Shi X (2002) RETRACTED: Ubiquitination and degradation of Cdc25C contribute to As(III)-induced G2/M cell cycle arrest. J Biol Chem. doi: 10.1074/jbc.M107813200