원문정보
초록
영어
In mammal, unfertilized oocytes remain in the oviduct or under in vitro culture, which is called "oocyte aging". This asynchrony negatively affects fertilization in pre- and post-implantation embryo development. Caffeine a phosphodiesterase inhibitor is known to rescue oocyte aging in several species. The objective of this study is to determine the cytoskeleton distribution in aged oocytes and the embryo developmental ability of aged oocytes in the present or absence of caffeine during maturation. Caffeine treatment increased the incidence of normal spindle assembly of aged oocytes (treatment, 67.57±4.11% aging, 44.61±6.4%) and no significant differences compared to control group. Fluorescence values were compared using ROS (Reactive oxidation species) stain. Fluorescence values appear of control group intensity rate (51.53.±3.80), aging group (68.10±5.54) and treatment of caffeine (45.04±2.98). Aged oocytes that were derived from addition of caffeine to the IVM (in vitro maturation) medium had significantly increased 2-cell that developed to the blastocyst stage compared to the aging group. Blastocysts, derived from caffeine treatment group, significantly increased the total cell number compare aging (90.44±10.18 VS 67.88±7.72). Apoptotic fragments of genomic DNA were measured in individual embryo using TUNEL assay. Blastocyst derived from caffeine treatment group decreased significantly the apoptotic index compared to blastocyst derived from aging group. In conclusion, we inferred that the caffeine treatment during oocyte aging can improve the developmental rate and quality in bovine embryos developing in vitro.
목차
INTRODUCTION
MATERIALS AND METHODS
Preparation of Bovine Oocytes and In Vitro Aging of Bovine Oocytes
In Vitro Fertilization of Bovine Oocytes and Embryo Culture
During In Vitro Maturation Treatment of Caffeine
Immunofluorescence and Measurement of ROS
Confocal Microscopy
Real Time Reverse Transcription Polymerase Chain Reaction(real time RT-PCR)
Statistical Analysis
RESULTS
Changes of Meiotic Resumption in Bovine Oocytes Aged For 42 h
Changes of Fertility and Development in Bovine Oocytes Aged for 42 h
Effect of Meiotic Resumption in Aging Bovine Oocytes with Caffeine Treatment
Effect of Fertility and Development in Aging Bovine Oocytes with Caffeine Treatment
Effect of Apoptosis in Bovine Oocytes Aged for 42 h with and without Caffeine Treatment
DISCUSSION
REFERENCES
