Jang, H.Y.
(Department of Genomic Engineering, Genomic Informatics Center, Hankyong National University)
,
Kong, H.S.
(Department of Genomic Engineering, Genomic Informatics Center, Hankyong National University)
,
Choi, K.D.
(Department of Genomic Engineering, Genomic Informatics Center, Hankyong National University)
,
Jeon, G.J.
(Department of Genomic Engineering, Genomic Informatics Center, Hankyong National University)
,
Yang, B.K.
(College of Animal Resources Science, Kangwon National University)
,
Lee, C.K.
(School of Agricultural Biotechnology, Seoul National University)
,
Lee, H.K.
(Department of Genomic Engineering, Genomic Informatics Center, Hankyong National University)
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 /in vitro fertilization were cultured in 5% $...
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 /in vitro fertilization were cultured in 5% $CO_2$ and 20% $O_2$ at $37^{\circ}C$ in NCSU23 medium. Melatonin was added to medium at concentration of 1nM, 5 nM, and 10 nM. When treated with 1nM (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${\pm}$0.5 and 17.3${\pm}$0.8), 1 nM (23.6${\pm}$0.6 and 19.0${\pm}$0.5), and 5 nM (23.3${\pm}$1.1 and 16.3${\pm}$0.8) treated with melatonin were higher than in 10 nM (20.0${\pm}$0.5 and 13.3${\pm}$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.
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 /in vitro fertilization were cultured in 5% $CO_2$ and 20% $O_2$ at $37^{\circ}C$ in NCSU23 medium. Melatonin was added to medium at concentration of 1nM, 5 nM, and 10 nM. When treated with 1nM (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${\pm}$0.5 and 17.3${\pm}$0.8), 1 nM (23.6${\pm}$0.6 and 19.0${\pm}$0.5), and 5 nM (23.3${\pm}$1.1 and 16.3${\pm}$0.8) treated with melatonin were higher than in 10 nM (20.0${\pm}$0.5 and 13.3${\pm}$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.
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문제 정의
The aim of the present study was to examine the effects of melatonin on the development and on antioxidant enzyme genes expression in vitro porcine embryos.
However, whether or not melatonin exerts its effects on embryo development in pigs has not been attempted. This study examined the effects of melatonin on the development and on expression of catalase, bax and caspase-3 genes expression in in vitro porcine embryos. We found that melatonin improved the development rate of morula plus blastocysts when embryos were cultured in NCSU23 medium containing melatonin, and the cell numbers of blatocysts produced by IVM/IVF were significantly increased in 1 nM and 5 nM treated with melatonin.
가설 설정
a,b Values with different superscripts within column are significantly differ. p<0.
a,b Values with different superscripts within column are significantly differ. p<0.
제안 방법
In the experiment, embryos were cultured under an atmosphere of 5% CO2 and 20% at 38.5℃ with different concentration (0, 1, 5 and 10 nM) of melatonin (Sigma, USA) for development.
데이터처리
The SAS mixed linear model program was used to analyze the data. Percentage of developmental stage was based upon the number of 2- to 8- cell embryos cultured in each treatment.
이론/모형
Treatment means were compared for differences through use of Duncan's Modified Multiple Range test.
Percentage of developmental stage was based upon the number of 2- to 8- cell embryos cultured in each treatment. Treatment means were compared for differences through use of Duncan's Modified Multiple Range test.
성능/효과
In conclusion, the present study demonstrates that addition of melatonin in culture medium to early pre-implantation porcine embryos enhances embryonic development. Although our data from porcine embryo development can not be directly applied to human fertilization and embryo transfer in vitro, melatonin might improve the culture conditions for such programs.
In conclusion, the present study demonstrates that addition of melatonin in culture medium to early pre-implantation porcine embryos enhances embryonic development. Although our data from porcine embryo development can not be directly applied to human fertilization and embryo transfer in vitro, melatonin might improve the culture conditions for such programs.
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