Effects of Melatonin on Gene Expression of IVM/IVF Porcine Embryos

The effect of melatonin on in vitro embryo development and the expression of antioxidant enzyme gene in preimplantation porcine embryos was determined by modified semi-quantitative single cell RT-PCR. Porcine embryos derived from in vitro maturation lin vitro fertilization were cultured in 5% CO2 and 20% O-2 at 37 degreesC in NCSU23 medium. Melatonin was added to medium at concentration of 1 nM, 5 nM, and 10 nM. When treated with 1 nM (39.0%) of melatonin, the developmental rate of embryos beyond the morula stage were higher than that of control group (31.0%) (p < 0.05). Number of inner cell mass and tropectoderm cell in control (23.0 +/- 0.5 and 17.3 +/- 0.8), 1 nM (23.6 +/- 0.6 and 19.0 +/- 0.5), and 5 nM (23.3 +/- 1.1 and 16.3 +/- 0.8) treated with melatonin were higher than in 10 nM (20.0 +/- 0.5 and 13.3 +/- 0.8) treated with melatonin (p < 0.05). To develop an mRNA phenotypic map for the expression of catalase, bax and caspase-3, single cell RT-PCR analysis were carried out in porcine IVM/IVF embryo. Catalase was detected in 0, 1 and 5 nM supplemented with melatonin, but bax and caspase-3 were detected in 10 nM treated with melatonin.