마우스 수정란에 있어서 부계 DNA 손상이 부계 DNA 퇴화 및 초기 배발달에 미치는 영향 = Effect of Paternal DNA Damage on Paternal DNA Degradation and Early Embryonic Development in Mouse Embryo: Supporting Evidence by GammaH2AX Expression

다국어 초록 (Multilingual Abstract)

This study was investigated to test whether the zygote recognized the topoisomerase II beta (TOP2B) mediated DNA fragmentation in epididymal spermatozoa or the nuclease degradation in vas deferens spermatozoa by testing for the presence of gammaH2AX (γH2AX). The γH2AX is phosphorylation of histone protein H2AX on serine 139 occurs at sites flanking DNA double-stranded breaks (DSBs). The presence of γH2AX in the pronuclei of mouse zygotes which were injected with DNA broke epididymal spermatozoa was tested by immunohistochemistry at 5 and 9 h post fertilization, respectively. Paternal pronuclei that arose from epididymal spermatozoa treated with divalent cations did not stain for γH2AX at 5 h. On the other hand, in embryos injected with vas deferences spermatozoa that had been treated with divalent cations, γH2AX was only present in paternal pronuclei, and not the maternal pronuclei at 5 h. Interestingly, both pronuclei stained positively for γH2AX for all treatments and controls at 9 h after sperm injection. In conclusion, the embryos recognize DNA that is damaged by nuclease, but not by TOP2B because H2AX in phosphorylated in paternal pronuclei resulting from spermatozoa treated with fragmented DNA from vas deferens spermatozoa treated with divalent cations, but not from epididymal spermatozoa treated the same way.

참고문헌 (Reference)

1 Shaman JA, "TopoisomeraseIIB and an extracellular nuclease interact to digest sperm DNA in an apoptotic-like manner" 75 : 741-748, 2006

2 Yamauchi Y, "Topoisomerase II-mediated breaks in spermatozoa cause the specific degradation of paternal DNA in fertilized oocytes" 76 : 666-672, 2007

3 Sirlin JL, "Timing of DNA synthesis in ovarian oocyte nuclei and pronuclei of the mouse" 18 : 190-194, 1959

4 Leno GH, "The nuclear membrane prevents replication of human G2 nuclei but not G1 nuclei in Xenopus egg extract" 69 : 151-158, 1992

5 Blow JJ, "Preventing re-replication of DNA in a single cell cycle: evidence for a replication licensing factor" 122 : 993-1002, 1993

6 Yamauchi YS, "Paternal pronuclear DNA degradation is functionally linked to DNA replication in mouse oocyte" 77 : 407-415, 2007

7 Gawecka JE, "Mouse zygotes respond to severe sperm DNA damage by delaying paternal DNA replication and embryonic development" 8 : e56385-, 2013

8 Ortega MA, "Mammalian sperm chromatin as a model for chromation funcion in DNA degradation and DNA replication" 57 (57): 43-49, 2011

9 Kimura Y, "Intracytoplasmic sperm injection in the mouse" 52 : 709-720, 1995

10 Redon CE, "Histone gammaH2AX and poly(ADP-ribose)as clinical pharmacodynamic biomarkers" 16 : 4532-4542, 2010

11 김창진, "Evidence of DNA Replication Licensing and Paternal DNA Degradation by MCM7 and ORC2 in the Mouse One-cell Embryo" 대한의생명과학회 23 (23): 372-379, 2017

12 Zielger-Birling C, "Distribution of p53 binding protein 1 (53BP1) and phosphorylated H2A.X during mouse preimplantation development in the absence of DNA damage" 53 : 1003-1011, 2009

13 Rogakou EP, "DNA double stranded breaks induce histone H2AX phosphorylation on serine 139" 273 : 5858-5868, 1998

14 Nashun B, "Changes in the nuclear deposition of histone H2A variants during preimplantation development in mice" 137 : 3785-3794, 2010

15 Blow JJ, "A role for the nuclear envelope in controlling DNA replication within the cell cycle" 332 : 546-548, 1988