The aim of this study was to determine if there are any associations between impaired chromatin packaging and the origin of reactive oxygen species (ROS) production. Cytosolic ROS, mitochondrial ROS, DNA protamination, and apoptosis were evaluated with dichlorofluoresceindiacetate (DCFH-DA), dihydrorhodamine 123 (DHR123), chromomycin A3 (CMA3), and YO-Pro-1 (Y1)/propidium iodide (PI), respectively, by flow cytometry (FCM) in 40 infertile individuals. Percentages DCF+ and R123+ sperm were positively associated with percentage CMA3+ sperm and negatively associated with percentage apoptotic sperm. No correlation was observed between CMA3+ sperm and the percentage of apoptotic sperm. Under protamination of sperm is not associated with the origin of ROS production, but their relationship may suggest an association with general physiological dysfunction of sperm. Furthermore, under protamination does make sperm prone to apoptosis. Rather, it is likely that apoptosis is induced by ROS production. Considering that these conclusions are derived from correlative analyses, additional studies including an interventive approach are required.

IntrOductIOnIVM of oocytes is an effective method for the production of mature oocyte used in ART, which includes IVF cloning and Intra Cytoplasmic Sperm Injection (ICSI). IVM is a cost-benefit technique with few side effects for gonadotropin stimulation of IVF, and is a simple treatment for certain infertile couple [1][2][3][4]. The GV-stage oocytes which are stripped of cumulus cells have a reduced developmental capacity when compared with cumulus enclosed GV-stage oocytes [5,6]. Cumulus cells play an important role in oocyte maturation, since they provide and transfer several known and unknown factors that are essential in the regulation of meiotic progression, normal nuclear and cytoplasmic maturation of oocytes and subsequent embryonic development after fertilization [7,8]. Studies have demonstrated that inappropriate culture mediums increase ROS resulting in OS events, DNA damage and low quality of oocytes. These are the main factors that lead to failure in IVM and decrease productivity [9,10]. OS appears to be responsible for many injuries in the embryo and oocyte [9,11,12]. Embryo metabolism generates ROS via several enzymatic mechanisms that penetrate the cell membrane and pass through it to damage cellular molecules such as lipids, proteins, and nucleic acids [13]. BME and CYS are thiol compounds that stimulate glutathione (GSH) synthesis [14]. Despite the fact that there is more stability of CYS than other thiol compounds, the effect of this substance on the growth of oocytes or embryos depending on breed, dose rate and the medium, is shown differently.GSH is one of the components of many biological processes, which includes the construction of DNA, protein metabolism of drugs and chemicals, and it protects the cells during OS events [15]. In addition, GSH and its revival mode of reproduction and early development of organism play a unique role. It can reduce the generation of free radicals such as hydrogen peroxide and oxygen free radicals, which are disruptive to oocytes. It has been shown that GSH plays an important role in oocyte maturation [15]. Intracellular GSH is an essential part in the development of the oocyte cytoplasm. Activities of GSH-related antioxidant properties protect the oocyte at the front of the active molecules of oxygen is toxic. With this consideration, this study was designed to evaluate the effects of antioxidants (BME and CYS) on improving ART invitro which is essential. Therefore, the main purpose of this study was to ascertain whether enriching the oocyte medium with antioxidants, BME and CYS may improve IVM and fertilization and embryo development into blastocyst, of mouse immature oocyte. MAterIAls And MethOdsThis experimental study was carried out in the cellular and molecular research center at Yasuj University of Medical Sciences in Iran. The University ethic committee approved the design of study. Sixteen mice were housed under standard laboratory conditions (temperature of 20±2°C, relative humidity of 40-45% and light-dark cycle of 12:12 hour) and had free access ...