Detection of reactive oxygen species (ROS) and apoptosis in human fragmented embryos

被引:395
作者
Yang, HW
Hwang, KJ
Kwon, HC [1 ]
Kim, HS
Choi, KW
Oh, KS
机构
[1] Ajou Univ, Sch Med, Dept Obstet & Gynecol, Div Reprod Endocrinol & Infertil, Suwon 442749, South Korea
[2] PL Infertil Clin, Seoul, South Korea
关键词
apoptosis; embryo fragmentation; reactive oxygen species;
D O I
10.1093/humrep/13.4.998
中图分类号
R71 [妇产科学];
学科分类号
100211 ;
摘要
In human in-vitro fertilization (IVF)-embryo transfer, the in-vitro culture environment differs from in-vivo conditions.at the oxygen concentration is higher, and in such conditions the mouse embryos show a higher concentration of reactive oxygen species (ROS) in simple culture media. ROS are believed to cause damage to cell membranes and DNA fragmentation in somatic cells. This study was conducted to ascertain the level of H2O2 concentration within embryos and the morphological features of cell damaged induced by H2O2. A total of 62 human oocytes and embryos (31 fragmented, 15 non-fragmented embryos, 16 unfertilized oocytes) was obtained from the IVF-embryo transfer programme, The relative intensity of H2O2 concentrations within embryos was measured using 2',7'-dichlorodihydrofluorescein diacetate by Quanti cell 500 fluorescence imaging and DNA fragmentation was observed with transmission electron microscopy and an in-situ apoptosis detection kit. The H2O2 concentrations were significantly higher in fragmented embryos (72.21 +/- 9.62, mean +/- SEM) compared to non-fragmented embryos (31.30 +/- 3.50, P < 0.05) and unfertilized oocytes (30.75 +/- 2.67, P < 0.05), Apoptosis was observed only in fragmented embryos, and was absent in non-fragmented embryos. microscopic findings confirmed apoptotic bodies and cytoplasmic condensation in the fragmented blastomeres. We conclude that there is a direct relationship between increased H2O2 concentration and apoptosis, and that further studies should be undertaken to confirm these findings.
引用
收藏
页码:998 / 1002
页数:5
相关论文
共 31 条
[1]   BENEFICIAL EFFECT OF EDTA ON DEVELOPMENT OF MOUSE ONE-CELL EMBRYOS IN CHEMICALLY DEFINED MEDIUM [J].
ABRAMCZUK, J ;
SOLTER, D ;
KOPROWSKI, H .
DEVELOPMENTAL BIOLOGY, 1977, 61 (02) :378-383
[2]   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
[3]  
ARENDS MJ, 1990, AM J PATHOL, V136, P593
[4]   COUPLED OXIDATION OF NADPH WITH THIOLS AT NEUTRAL PH [J].
BACCANARI, DP .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1978, 191 (01) :351-357
[5]   DIRECT DEMONSTRATION OF SUPEROXIDE ANION PRODUCTION DURING OXIDATION OF REDUCED FLAVIN AND OF ITS CATALYTIC DECOMPOSITION BY ERYTHROCUPREIN [J].
BALLOU, D ;
PALMER, G ;
MASSEY, V .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1969, 36 (06) :898-&
[6]   EFFECT OF OXYGEN CONCENTRATION ON IN-VITRO FERTILIZATION AND EMBRYO CULTURE IN THE HUMAN AND THE MOUSE [J].
DUMOULIN, JCM ;
VANVUCHELEN, RCM ;
LAND, JA ;
PIETERS, MHEC ;
GERAEDTS, JPM ;
EVERS, JLH .
FERTILITY AND STERILITY, 1995, 63 (01) :115-119
[7]  
ERENUS M, 1991, FERTIL STERIL, V56, P707
[8]  
GAIDO ML, 1991, J BIOL CHEM, V266, P18580
[9]   INCREASED GENERATION OF REACTIVE OXYGEN SPECIES IN EMBRYOS CULTURED INVITRO [J].
GOTO, Y ;
NODA, Y ;
MORI, T ;
NAKANO, M .
FREE RADICAL BIOLOGY AND MEDICINE, 1993, 15 (01) :69-75
[10]   OXIDATIVE STRESS ON MOUSE EMBRYO DEVELOPMENT INVITRO [J].
GOTO, Y ;
NODA, Y ;
NARIMOTO, K ;
UMAOKA, Y ;
MORI, T .
FREE RADICAL BIOLOGY AND MEDICINE, 1992, 13 (01) :47-53