ABSTRACT
Background: Prenatal drug exposure, as a common public health concern, is associated with an increased risk of adverse effects on early embryo development
Objective: To investigate the in vitro development of - embryo from experimentally Kerack-addicted mice
Materials and Methods: Twenty-five female mice were studied in five groups: control, vehicle, and three experimental groups of Kerack-dependent mice [I, II, and III] which received different doses of Kerack for 14 days. After the establishment of addiction model [7 days], experimental groups I, II, and III were given Kerack intraperitoneally at the doses of 5, 35, and 70 mg/kg, twice a day for a period of 7 days, respectively. The vehicle group received normal saline and lemon juice whilst the control group just received water and food. Morulae were obtained through oviduct flashing. The survived embryos were cultured in T6+ 5mg/ml bovine serum albumin. The developmental rates up to hatched stage daily and embryo quality [differential staining and Tunnel staining] were also assessed
Results: The developmental potential of embryos obtained from the addicted mother was significantly decreased in comparison with control group. There was a significant reduction in the rate of blastocyst formation in the high dose Kerack dependent group. However, in addicted mice there was reduction in the total cell number [40.92% vs. 65.08% in control] and, inner cell mass percentage [17.17% vs. 26.15% in control] while apoptotic cells numbers were increased [7.17 vs. 1.46 in control] [p<0.05]
Conclusion: The Kerack addiction during pregnancy retards preimplantation development and induces apoptosis
Subject(s)
Animals, Laboratory , Substance-Related Disorders , In Vitro Techniques , Mice , ApoptosisABSTRACT
The objective of this study was to investigate the effect of vitrification and in vitro maturation on the mitochondrial distribution and ATP content of oocytes. The oocytes at Germinal Vesicle [GV] and Metaphase II [MII] stages were recovered from 6-8 week old NMRI strain female mice. The oocytes were divided into vitrified and non-vitrified groups. Vitrification was done by the cryotop method using ethylene glycol, dimethylsulfoxide and sucrose as cryoprotectants. The GV oocytes were cultured in maturation medium for 24 hrs. The collected in vitro matured oocytes [IVM-MII] and ovulated metaphase II [OV-MII] oocytes were inseminated with capacitated sperm. The ATP content of the oocytes was measured by luciferin-luciferase reaction. Distribution of oocyte mitochondria was studied using Mito Tracker Green staining under fluorescent microscope. The survival rates of vitrified oocytes at GV and MII stages were 87.39 and 89.5%, respectively. There was no significant difference in the developmental and hatching rates of vitrified and non-vitrified oocytes. The ATP content of GV and MII oocytes derived from in vivo and in vitro condition was not significantly different in vitrified and non-vitrified samples. The pattern of mitochondrial distribution in vitrified and non-vitrified GV and MII oocytes was similar but it was different between MII oocytes collected from fallopian tube and in vitro matured MII oocytes. However, the florescent intensity of mitochondrial staining was different in all the groups in the study. Vitrification did not affect mouse oocyte developmental competence, ATP content at different developmental stages but some alteration was seen in mitochondria distribution of in vitro matured oocytes in comparison to their controls