Paternal exposure to cyclophosphamide induces DNA damage and alters the expression of DNA repair genes in the rat preimplantation embryo

被引:70
作者
Harrouk, W
Codrington, A
Vinson, R
Robaire, B
Hales, BF
机构
[1] McGill Univ, Dept Pharmacol, Montreal, PQ H3G 1Y6, Canada
[2] McGill Univ, Dept Therapeut, Montreal, PQ H3G 1Y6, Canada
[3] McGill Univ, Dept Obstet & Gynecol, Montreal, PQ H3G 1Y6, Canada
来源
MUTATION RESEARCH-DNA REPAIR | 2000年 / 461卷 / 03期
基金
英国医学研究理事会;
关键词
spermatozoa; comet assay; nucleotide excision repair; base excision repair; mismatch repair; cyclophosphamide; rat preimplantation embryo; gene expression;
D O I
10.1016/S0921-8777(00)00053-7
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Chronic low dose treatment of male rats with cyclophosphamide, an anticancer alkylating agent, damages male germ cells, resulting in greater than 80% peri-implantation progeny loss. Little transcription or repair takes place in the DNA of post-meiotic male germ cells. The spermatozoal genome regains its transcriptional capacity in the fertilized oocyte. We hypothesized that as a consequence of exposure of male rats to cyclophosphamide DNA damage to the male genome is transmitted to the conceptus; furthermore. this damage leads to alterations in the expression profiles of DNA repair genes during preimplantation development. Male rats were treated with either saline or cyclophosphamide (6 mg/kg/day, 4-6 weeks) and mated to control females; 1-8 cell stage embryos were collected. The alkaline comet assay was used to assess DNA damage in 1-cell embryos. A significantly higher percentage (68%) of the embryos fertilized by cyclophosphamide-exposed spermatozoa displayed a comet indicative of DNA damage, compared to those sired by control males (18%). The in situ transcription/antisense RNA approach was used to determine if DNA damage alters the expression of DNA repair genes in early embryos. Dramatic increases in the transcripts for selected members of the nucleotide excision repair family (XPC, XPE and PCNA), mismatch repair family (PMS1), and recombination repair family (RAD50) were found in 1-cell stage embryos shed by cyclophosphamide-treated males compared to controls, while decreases in the expression of base excision repair family members (UNG1, UNG2, and XRCC1) and in recombination repair transcripts (RAD54) were observed. By the 8-cell stage, transcripts for specific members of the nucleotide excision repair family (XPC) and mismatch repair family (MSH2, PMS2) were elevated greatly in control embryos compared to embryos sired by drug-treated males; in contrast, transcripts for other members of the nucleotide excision repair family (XPE, PCNA), as well as some of the base excision repair family (UNG1), were elevated in embryos sired by drug-treated males. Therefore, DNA damage incurred in spermatozoa, following cyclophosphamide exposure is associated with alterations in the expression profiles of DNA repair genes in preimplantation embryos as early as the I-cell stage. Genotoxic stress may disturb the nuclear remodeling and reprogramming events that follow fertilization and precede zygotic genome activation. (C) 2000 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:229 / 241
页数:13
相关论文
共 55 条
[41]  
1::AID-MRD1&gt
[42]  
3.0.CO
[43]  
2-0
[44]  
TAYLOR LE, 1995, J CRAN GENET DEV BIO, V15, P13
[45]  
TEBBS RS, 1998, REPROD DEV TOXICOLOG, P113
[46]   The role of homologous recombination processes in the repair of severe forms of DNA damage in mammalian cells [J].
Thacker, J .
BIOCHIMIE, 1999, 81 (1-2) :77-85
[47]   A TIME-COURSE STUDY OF CHRONIC PATERNAL CYCLOPHOSPHAMIDE TREATMENT IN RATS - EFFECTS ON PREGNANCY OUTCOME AND THE MALE REPRODUCTIVE AND HEMATOLOGIC SYSTEMS [J].
TRASLER, JM ;
HALES, BF ;
ROBAIRE, B .
BIOLOGY OF REPRODUCTION, 1987, 37 (02) :317-326
[48]   CHRONIC LOW-DOSE CYCLOPHOSPHAMIDE TREATMENT OF ADULT MALE-RATS - EFFECT ON FERTILITY, PREGNANCY OUTCOME AND PROGENY [J].
TRASLER, JM ;
HALES, BF ;
ROBAIRE, B .
BIOLOGY OF REPRODUCTION, 1986, 34 (02) :275-283
[49]   PATERNAL CYCLOPHOSPHAMIDE TREATMENT OF RATS CAUSES FETAL LOSS AND MALFORMATIONS WITHOUT AFFECTING MALE-FERTILITY [J].
TRASLER, JM ;
HALES, BF ;
ROBAIRE, B .
NATURE, 1985, 316 (6024) :144-146
[50]   PCNA, a multifunctional ring on DNA [J].
Tsurimoto, T .
BIOCHIMICA ET BIOPHYSICA ACTA-GENE STRUCTURE AND EXPRESSION, 1998, 1443 (1-2) :23-39