The effects of vitrification on gene expression in mature mouse oocytes by nested quantitative PCR.
OBJECTIVE
this study was conducted on the effects of vitrification cryotop method on gene expression of mature oocytes in Mus musculus.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Habibi
Afrooz
A
Cellular and Molecular Biology Research Center, Medical School of Shaheed Beheshti University of Medical Sciences and Health Services, Parvaneh St. Yaman St. Chamran highway, Tehran, Iran. Habibi_af@yahoo.com
Farrokhi
Naser
N
Moreira da Silva
Fernando
F
Bettencourt
Bruno F
BF
Bruges-Armas
Jácome
J
Amidi
Fardin
F
Hosseini
Ahmad
A
D000818:Animals; D000941:Antigens / Q000235:genetics / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology; D004527:Egg Proteins / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D051379:Mice; D008813:Mice, Inbred ICR; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D009865:Oocytes / Q000187:drug effects*; D016133:Polymerase Chain Reaction; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism; D000072105:Superoxide Dismutase-1
D000941:Antigens; D003451:Cryoprotective Agents; D004527:Egg Proteins; C471056:Hook1 protein, mouse; D008869:Microtubule-Associated Proteins; C416503:Nalp5 protein, mouse; C000606291:Sod1 protein, mouse; D013482:Superoxide Dismutase; D000072105:Superoxide Dismutase-1