GSH Level, Spindle Area andRate of IVF in Presence ofAntioxidant in Mouse OocyteCultured in MEME Medium


Mohammadi Roushandeh A *, Pasbakhsh P , Shokrgozar M.A , Alizadeh Z , Mahmoudi R , Habibi Roudkenar M ,

0

Introduction: The study was carried out effects of different doses of cysteamine on rate of IVM and IVF. Also the study was carried out to study glutathione (GSH) synthesis in presence of cysteamine without cumulus cells in MEME medium. MII spindle area was analysed by immunocytochemistry for quantification of shape and size of it. Materials and Methods: Female mice were primed with 5 IU of PMSG and GV oocytes were retrieved from the ovary 48 hr later for IVM. The IVM medium was supplemented with 0, 50, 100, 200 and 500 mM of cysteamine. Experiments also included a group of ovulated oocytes (in vivo matured) after priming with PMSG and HCG. Cytoplasmic GSH level was measured by DTNB-GR recycling protocol. For IVF MII oocytes were inseminated with mature mouse sperm and 24 hours after insemination rate of two cell embryo was measured. MII Oocytes were fixed and immunostained for microtubules, and chromosomes and then spindle area were analysed. Results: After IVM, an improvement was observed on MII development in 200 μm cysteamine group. Intracytoplasmic GSH level increased in presence of cysteamine in 200μm cysteamine and Highest level of GSH was produced in In vivo. Spindle area in all in vitro groups exept 500μm increased and Spindle area in 200μm cysteamine compare to in vivo group was insignificant(p>0.05). Conclusion: Our results showed that cysteamine improved IVM rate in dose dependant. Rate of two cell embryo increased significantly in 200μm cysteamine compare to control. Also cysteamine induced glutathione synthesis in MII oocyte and improved microtubule organization in 200μm cysteamine group.