A comparative study of oxidative DNA damage in mammalian spermatozoa

被引:128
作者
Bennetts, LE
Aitken, RJ [1 ]
机构
[1] Univ Newcastle, Sch Life & Environm Sci, Discipline Biol Sci, Callaghan, NSW 2308, Australia
[2] Univ Newcastle, ARC Ctr Excellence Biotechnol & Dev, Callaghan, NSW 2308, Australia
[3] Univ Newcastle, Hunter Med Res Inst, Callaghan, NSW 2308, Australia
关键词
DNA damage; protamine; oxidative stress; mitochondrial DNA; spermatozoa; tammar wallaby;
D O I
10.1002/mrd.20285
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 [生物化学与分子生物学]; 081704 [应用化学];
摘要
Alkaline gel electrophoresis, pulsed field gel electrophoresis, and quantitative PCR analyses (QPCR) of the nuclear (nDNA) and mitochondrial (mtDNA) genomes were used to assess DNA integrity in the spermatozoa of three species exposed to oxidative stress. In human and murine spermatozoa, the mtDNA was significantly more susceptible to H2O2-mediated damage than nDNA. In both eutherian species, exposure to 250 mu M H2O2 induced around 0.6 lesions/10 kb of mtDNA. The mtDNA of human spermatozoa was particularly vulnerable to oxidative stress; 0.25, 1, and 5 mM H2O2 inducing DNA damage equivalent to 0.62, 1.34, and 1.42 lesions/ 10 kb, respectively. Such results emphasize the diagnostic significance of mtDNA as a biomarker of oxidative stress in the male germ line. In contrast, no damage could be detected by QPCR in the nDNA of either eutherian species, on exposure to H2O2 at doses as high as 5 mM. However, electrophoretic analysis indicated that severe oxidative stress could induce detectable nDNA fragmentation in human, but not murine spermatozoa. The mtDNA of tammar wallaby spermatozoa was relatively resistant to oxidative stress, only exhibiting damage (0.6 lesions/10 kb DNA) on exposure to 5 mM H2O2. By contrast, the nDNA of wallaby spermatozoa was significantly more susceptible to this oxidant than the other species. Such vulnerability is consistent with the lack of disulfide cross-linking in marsupial sperm chromatin and suggests that chromatin condensation during epididymal maturation may be important in establishing the resistance of these cells to the genotoxic effects of reactive oxygen species. (c) 2005 Wiley-Liss, Inc.
引用
收藏
页码:77 / 87
页数:11
相关论文
共 101 条
[1]
REACTIVE OXYGEN SPECIES GENERATION AND HUMAN SPERMATOZOA - THE BALANCE OF BENEFIT AND RISK [J].
AITKEN, J ;
FISHER, H .
BIOESSAYS, 1994, 16 (04) :259-267
[2]
Aitken RJ, 1998, J CELL SCI, V111, P645
[3]
Aitken RJ, 1999, J REPROD FERTIL, V115, P1
[4]
Oxidative stress, DNA damage and the Y chromosome [J].
Aitken, RJ ;
Krausz, C .
REPRODUCTION, 2001, 122 (04) :497-506
[5]
AITKEN RJ, 1995, J CELL SCI, V108, P2017
[6]
Oxidative stress and male reproductive biology [J].
Aitken, RJ ;
Baker, MA .
REPRODUCTION FERTILITY AND DEVELOPMENT, 2004, 16 (05) :581-588
[7]
Relative impact of oxidative stress on the functional competence and genomic integrity of human spermatozoa [J].
Aitken, RJ ;
Gordon, E ;
Harkiss, D ;
Twigg, JP ;
Milne, P ;
Jennings, Z ;
Irvine, DS .
BIOLOGY OF REPRODUCTION, 1998, 59 (05) :1037-1046
[8]
GENERATION OF REACTIVE OXYGEN SPECIES, LIPID-PEROXIDATION, AND HUMAN-SPERM FUNCTION [J].
AITKEN, RJ ;
CLARKSON, JS ;
FISHEL, S .
BIOLOGY OF REPRODUCTION, 1989, 41 (01) :183-197
[9]
AITKEN RJ, 1988, J ANDROL, V9, P367
[10]
AITKEN RJ, 1987, J REPROD FERTIL, V81, P459