The effects of vitrification on gene expression in mature mouse oocytes by nested quantitative PCR

被引:26
|
作者
Habibi, Afrooz [1 ]
Farrokhi, Naser [2 ]
da Silva, Fernando Moreira [3 ]
Bettencourt, Bruno F. [4 ]
Bruges-Armas, Jacome [4 ]
Amidi, Fardin [5 ]
Hosseini, Ahmad [1 ]
机构
[1] Shaheed Beheshti Univ Med Sci & Hlth Serv, Sch Med, Cellular & Mol Biol Res Ctr, Tehran, Iran
[2] Shahrood Univ Technol, Dept Agron & Plant Breeding, Fac Agr, Shahrood, Iran
[3] Univ Azores, Dept Agr Sci, Angra Do Heroismo, Portugal
[4] Hosp Santo Espirito Angra Heroismo, SEEBMO, Angra Do Heroismo, Azores, Portugal
[5] Univ Tehran Med Sci, Sch Med, Dept Anat, Tehran, Iran
关键词
Cryotop; Gene expression; Nested quantitative; PCR (nqPCR); Vitrified oocyte (MII); EARLY EMBRYONIC-DEVELOPMENT; MATER; PROFILES; PATTERNS; MICE; DNA;
D O I
10.1007/s10815-010-9453-0
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
This study was conducted on the effects of vitrification cryotop method on gene expression of mature oocytes in Mus musculus. Transcript analyses of three mouse genes, namely Mater, Hook1 and Sod1, were performed upon non-vitrified and vitrified oocytes with different concentrations of dimethyl sulfoxide (DMSO) and ethylene glycol (EG),15%: 7.5% DMSO + 7.5% EG, and 30%: 15% DMSO + 15% EG, using cryotop following normalization of transcripts with Hprt1 by nested quantitative PCR. Vitrification caused down-regulation of Mater and Hook1 and up-regulation of Sod1 when lower concentrations of cryoprotectants were used as opposed to the control group. The relative expression of Sod1 in vit(2) (30% v/v) was significantly higher than vit(1) (15% v/v)(.) Quantitative transcript analysis of Mater and Hook1 for the vit(2) condition failed to produce any data. Survival rates were the same for both vitrification treatments and significantly lower than control group. Although vit(1) treatment had lower survival rate compared to control group, it demonstrated better stability comparing to vit(2) based on the transcript analysis.
引用
收藏
页码:599 / 604
页数:6
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