Animal physiology
Hamed Daneshpazhouh; Nasim Hayati Roodbari; Mehdi Dianatpour; Yaser Tahamtani; zahra khodabandeh
Abstract
Freezing is a long-term egg storage method that plays an important role in assisted reproductive methods. The aim of the present study is to investigate the effect of freezing solutions and docetaxel on the expression changes of autophagy genes such as Atg5 and Beclin-1 in mouse MII oocytes after glass ...
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Freezing is a long-term egg storage method that plays an important role in assisted reproductive methods. The aim of the present study is to investigate the effect of freezing solutions and docetaxel on the expression changes of autophagy genes such as Atg5 and Beclin-1 in mouse MII oocytes after glass freezing by cryotop method. To achieve this goal, mouse MII oocytes were collected and frozen in two different concentrations of 15% ethylene glycol, 15% dimethyl sulfoxide and 0.5 M sucrose in group A (VS1) and 7.5% ethylene glycol, glycerol. 7.5% and 0.5 M sucrose were frozen in group B (VS2) and some groups were affected by docetaxel before freezing. After thawing, the eggs were fertilized.The percentage of survival and fertilization of frozen and thawed oocytes was evaluated and the expression changes of genes (Atg5 and Beclin-1) were investigated by RT-PCR method. The results showed that there are significant differences between the percentage of survival and the percentage of fertilization in the freezing groups compared to the control group (P<0.05). The percentage of survival and fertilization in the VS1 group decreased compared to the VS2 group. Also, the percentage of survival and conception of the groups pre-incubated with Docetaxel was higher than the non-incubated groups. This study showed that vitrification with cryotop changes the transcript levels of autophagy genes in frozen-thawed MII oocytes, and pre-incubation of oocytes with docetaxel before vitrification can decrease the transcript levels of Atg5 and Beclin-1 in the experimental groups and above Increase the percentage of survival and the percentage of formation of two-celled embryos.
Naeimeh Dehghani; Mehdi Dianatpour; Seyed Ebrahim Hosseini; Zahra Khodabandeh; Hamed Daneshpazhouh
Abstract
As a stabilizing agent, docetaxel can potentially reduce the damage to the oocyte cytoskeleton during vitrification. The aim of the present study was to investigate the effect of docetaxel on the survival rate and in vitro fertilization of oocytes after vitrification. NMRI mice (8-10 weeks old) were ...
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As a stabilizing agent, docetaxel can potentially reduce the damage to the oocyte cytoskeleton during vitrification. The aim of the present study was to investigate the effect of docetaxel on the survival rate and in vitro fertilization of oocytes after vitrification. NMRI mice (8-10 weeks old) were superovulated by injecting PMSG and HCG. Oocytes are surrounded by cumulus and corona cells and must be denuded by 0.1% hyaluronidase enzyme. The oocytes were then divided into 5 experimental groups including control, docetaxel, docetaxel+vitrification solution; docetaxel+ vitrification and vitrification. Mature oocytes were vitrified in ethylene glycol and dimethyl sulfoxide solutions at 15% concentration and 0.5 M sucrose. After thawing, their survival and fertilization rates were assessed up to the two-cell stage. Staining of the microtubules in the oocytes was performed with alpha-tubulin antibody. The fertilization rate of each group showed a significant decrease compared to the control group (P=0.001). The rate of formation of 2-cell embryos in both vitrified groups (docetaxel+ vitrified and vitrified vitrified) was significantly lower than non-vitrified (control (P=0.001) and docetaxel ((P=0.004)). The results showed that survival and fertilization rates in pre-incubated groups with docetaxel were higher than non-incubated groups, so docetaxel could improve reproductive techniques by reducing the damage to the oocyte cytoskeleton.
