Open AccessPaternal DNA Damage Resulting From Various Sperm Treatments Persists After Fertilization and Is Similar Before and After DNA Replication
Author(s) -
Yamauchi Yasuhiro,
Riel Jonathan M.,
Ward Monika A.
Publication year - 2012
Publication title -
journal of andrology
Language(s) - English
Resource type - Journals
eISSN - 1939-4640
pISSN - 0196-3635
DOI - 10.2164/jandrol.111.013532
Subject(s) - dna damage , sperm , biology , comet assay , dna fragmentation , chromatin , human fertilization , intracytoplasmic sperm injection , dna , andrology , dna replication , microbiology and biotechnology , genetics , embryo , in vitro fertilisation , medicine , programmed cell death , apoptosis
In spite of its highly condensed state, sperm DNA is vulnerable to damage that can originate from oxidative stress, the activity of sperm‐specific nucleases, or both. After fertilization, in the oocyte, paternal chromatin undergoes dramatic changes, and during this extensive remodeling, it can be both repaired and degraded, and these processes can be linked to DNA synthesis. Here, we analyzed sperm response to damage‐inducing treatments both before and after fertilization and before or after zygotic DNA replication. Epididymal mouse spermatozoa were either frozen without cryoprotection (FT) or treated with detergent Triton X‐100 coupled with dithiothreitol (TX+DTT) to induce DNA damage. Fresh, untreated sperm served as control. Immediately after preparation, spermatozoa from 3 groups were taken for comet assay, or for intracytoplasmic sperm injection into prometaphase I oocytes to visualize prematurely condensed single‐chromatid chromosomes, or into mature metaphase II oocytes to visualize chromosomes after DNA replication. Comet assay revealed increased DNA fragmentation in treated sperm when compared with control, with FT sperm more severely affected. Chromosome analysis demonstrated paternal DNA damage in oocytes injected with treated, but not with fresh, sperm, with FT and TX+DTT groups now yielding similar damage. There were no differences in the incidence of abnormal paternal karyoplates before and after DNA synthesis in all examined groups. This study provides evidence that subjecting sperm to DNA damage–inducing treatments results in degradation of highly condensed sperm chromatin when it is still packed within the sperm head, and that this DNA damage persists after fertilization. The difference in DNA damage in sperm subjected to 2 treatments was ameliorated in the fertilized oocytes, suggesting that some chromatin repair might have occurred. This process, however, was independent of DNA synthesis and took place during oocyte maturation.