• Development and Reproduction
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• v.12 no.1
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• pp.19-30
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• 2008

Proper development of fertilized oocyte to blastocyst is a key step in mammalian development to implantation. During development of preimplantation embryos, the mammalian embryo needs supply the energy substrate for keep viability. Usually mammalian oocyte get substrate especially energy substrate from oviduct and uterus, because it does not store much substrate into cytoplasm during oogenesis. Carbohydrates are known as a main energy substrate for preimplantation stage embryos. Glucose, lactate and pyruvate are essential component in preimplantation embryo culture media and there are stage specific preferences to them. Glucose transporter and $H^+$-monocarboxylate cotransporter are a main mediator for carbohydrate transport and those expression levels are primarily under the control of intrinsic or extrinsic factors like insulin and glucose. Other organic substances, amino acids, lipids and nucleotides are used as energy substance and cellular regulation factor. Though since 1960s, successful development of fertilized embryo to blastocyst has been accomplished with chemically defined medium for example BWW and give rise to normal offspring in mammals, the role of metabolites and the regulation of intermediary metabolism are still poorly understood. Glucose may permit expression of metabolic enzymes and transporters in compacting morula, capable of generating the energy required for blastocyst formation. In addition, it has been suggested that the cytokines can modulate the metabolic rate of carbohydrate in embryos and regulate the preimplantation embryonic development through control the metabolic rate. Recently we showed that lactate can be used as a mediator for preimplantation embryonic development. Those observations indicate that metabolites of carbohydrate are required by the early embryo, not only as an energy source, but also as a key substrate for other regulatory and biosynthetic pathways. In addition metabolites of carbohydrate may involve in cellular activity during development of preimplantation embryos. It is suggested that through these regulation and with other regulation mechanisms, embryo and uterus can prepare the embryo implantation and further development, properly.

• 대한생식의학회:학술대회논문집
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• 2002.11a
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• pp.57.1-57.1
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• 2002

• Development and Reproduction
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• v.4 no.1
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• pp.29-35
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• 2000

A phosphatidylinositol 3-kinase (PI3K) is a upstream component of insulin signaling by which protein synthesis can be stimulated in many systems. To elucidate involvement of PI3K and its downstream mammalian target of rapamycin (mTOR) in the insulin signaling in pleimplantation mouse embryos, 8-cell embryos were cultured to blastocysts in the presence or absence of insulin and／or inhibitor drugs. The number of blastomeres per blastocyst, protein synthesis, and protein phosphorylation were examined. There was significant difference in embryonic development to blastocyst stage and hatching was potentiated by the insulin supplementation. The increase in the mean celt numbers per blastocyst was apparent in the insulin culture. Wortmannin, a PI3K inhibitor and rapamycin, an inhibitor of mTOR abolished the stimulatory effect of insulin on morphological development mitosis and protein synthesis. In autoradiography, phosphoproteins pp22 and pp30 which undergo phosphorylation in response to insulin were identified. Taken together, it can be suggested that PI3K and mTOR engaged in insulin signaling in the mouse embryo 8-cell onward and mediate embryotropic offset of insulin.

• Korean Journal of Environmental Biology
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• v.30 no.2
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• pp.128-135
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• 2012

The individual toxicity of lead (Pb) and zinc (Zn) has been investigated by using the sea urchin (Hemicentrotus pulcherrimus) germ cell and pluteus-larvae. The gametotoxic and embryotoxic effects of Pb and Zn on H. pulcherrimus were each investigated at 31, 63, 125, 250, 500 ppb and 16, 31, 63, 125, 250 ppb, respectively. Spawning was induced by 0.5 M KCl solution and the fertilization and normal embryogenesis rates test were performed for 10 min and 64 h after fertilization, respectively. In exposure to Pb, the fertilization rate was not significantly changed compared with control but normal embryogenesis rate was significantly decreased with concentration dependent manner. Fertilization and normal embryogenesis rates showed a significant decreased with concentration dependent manner in exposed to Zn. The normal embryogenesis rates were significantly inhibited in exposed to Pb ($EC_{50}$=45.13 ppb, 95% Cl=40.12~50.05 ppb) and Zn ($EC_{50}$=19.82 ppb, 95% Cl=18.26~21.31 ppb). In exposure to Pb and Zn, the NOEC of normal embryogenesis rate was <31.25 and <15.63 ppb, respectively. The LOEC showed each 31.25 and 15.63 ppb in exposed to Pb and Zn. These results suggest that the early embryo development of H. pulcherrimus is highly sensitive to heavy metals such as Pb and Zn, H. pulcherrimus can be used as a test organism for risk assessment in marine ecosystems.

• Korean Journal of Environment and Ecology
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• v.18 no.3
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• pp.340-345
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• 2004

The embryotoxic effects of PCB (Aroclor 1248) on a Korean frog, Rana dybowskii was deter-mined by using the FETAX (Frog Embryo Teratogenesis Assay-Xenopus) protocol. The rates of mortality and malformed larvae were investigated by probit analysis. The results showed that PCB is highly embryolethal. From LC$_{50}$ of 1.48ppb and from EC$_{50}$ of 0.25ppb and TI of 5.7 were derived, which indicates PCB is to be considered a teratogenic compound. Specific malformations occurred in 62.0% as edema at the 0.1ppb, in 32.0% as tail deformations at the 1ppb, and in 68.0% as profound deformations at the 5ppb of PCB concentration which living embryos were exposed to. PCB suppressed the growth of head-tail length at a relatively low concentration (1.0ppb), and therefore growth inhibition as assessed by embryo length can be used as a sensitive indicator to evaluate the toxicity of pollutants in the environment. In conclusion, PCB must be considered highly embryotoxic to Rana dybowskii.

• The Science & Technology
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• v.34 no.9 s.388
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• pp.43-68
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• 2001

인간배아란 무엇인가 - 조직분화단계까지의 세포집단이 대상, 연구 아직 초보단계 안전성 확보가 관건/인간배아 연구 찬성한다 - 질병발생기전 이해 난치병 극복 길 열어, 인간 삶의 질 향상이 목표 연구허용 마땅/인간배아 연구 반대한다 - 수정란 자체가 생명체 파괴 있을 수 없어, 인간존엄성 어느 누구도 해 입힐 수 없다/인간배아 연구와 윤리 - 인간조재ㆍ정체성 혼란에 불안과 두려움, 과학과 종교ㆍ인문사회계 견해 차 좁혀야/주요 선진국 연구동향 - 부시 미 연방정부 예산 지원 발표 큰 파문, 영국 가장 개방적, 독일은 일체 연구금지

• Journal of the korean veterinary medical association
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• v.41 no.10
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• pp.918-928
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• 2005

체세포 핵이식을 통한 정상인과 환자 체세포 유래의 다능성 인간배아 줄기세포주 확립 및 응용 - 체세포 핵이식을 통한 다능성 인간 배아 줄기세포주 확립의 기술적 측면 - 인간배아줄기세포의 분화 : 발생생물학적 접근 및 세포치료에 대한 전망 - 마우스 배아줄기세포 기능들의 호르몬 조절 - 간엽줄기세포를 이용한 골조직 공학 - 조혈모세포의 재생 - 제대혈 유래 간엽줄기세포를 이용한 세포치료 - 난치성 혈액종양질환에서 수지상세포를 이용한 세포면역치료법 확립

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2001.08a
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• pp.45-47
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• 2001

생쥐 착상전 초기배아에서 insulin과 tumor necrosis factor $\alpha$ (TNF$\alpha$)에 의한 배아의 형태 발생, 세포증식, apoptosis 및 MAPK활성의 변화를 조사하였다. Insulin에 의해 형태발생 및 포배당 세포수가 증가되었으며 TNF $\alpha$ 처리시 유의하게 감소하였다. TNT$\alpha$ 전처리시 insulin에 의한 발생 및 세포수 증가 촉진효과가 상쇄되었으며 TNF$\alpha$는 배아내 caspase-3의 활성을 증가시켰다. Insulin은 단시간내에 포배에서 mitogen activated protein kinase (MAPK or Erk1/2)의 활성을 증가시켰다. TNF$\alpha$는 포배내 MAPK의 활성을 감소시켰다. Insulin 처리 전 TNF $\alpha$를 전처리한 경우 insulin에 의한 MAPK 활성의 증가가 상쇄되었다. 배발생을 촉진하는 인슐린 신호전달 과정은 MAPK cascade를 경유하며 TNF $\alpha$를 경유한 신호전달과 Crosstalk이 존재하는 것으로 사료된다.

• Development and Reproduction
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• v.10 no.2
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• pp.105-113
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• 2006

Reactive oxygen species(ROS) generated in cellular metabolism have an effect on cell maturation and development. In human reproductive tract, oxidative injury by ROS may induce female infertility. Also, oxidative injury may be responsible for developmental retardation and arrest of mammalian preimplantation embryos. Activating transcription factor 4(ATF4) is a member of the cyclic-AMP response element-binding(CREB) familiy of basic region- leucine zipper(bZip). ATF4 is known to regulate stress response to protect cell from various stress factors and inducer of apoptisis. The purpose of this study was to investigate whether ATF4 is involved in the defensive mechanism in oxidative stress condition during the development of mouse preimplantation embryos. To verify the expression of ATF4 in oxidative stress condition, 2-cell stage embryos were cultured in HTF media containing 0.1mM, 0.5mM or 1mM hydrogen peroxide($H_2O_2$) for 1hr(2-cell), 8hr(4-cell), 17hr(8-cell), 24hr(morula), 48hr(early blastocyst) or 64hr(late blastocyst). The developmental rate decreased in the 0.1mM $H_2O_2$ treated group compared with control group. In embryos treated with 0.5mM and 1mM $H_2O_2$ showed 2-cell block. As a results of the semi-quantitative RT-PCR analysis of SOD1, ATF4 and Bax gene expression, SOD1, ATF4 and Bax genes were increased in 0.1mM, 0.5mM, 1mM $H_2O_2$ treated groups compared with control group. In 2-cell embryos, expression of SOD1, ATF4 and Bax genes were notably increased in 0.1mM, 0.5mM, 1mM $H_2O_2$ treated groups compared with control group. Immunofluorescence analysis showed that ATF4 protein was localized at the cytoplasm of preimplantation embryos. The increase in ATF4 immunoreactivety was observed in the 0.1mM, 0.5mM, 1mM $H_2O_2$ treated groups compared with control group. It suggests that oxidative stress by $H_2O_2$ induces expression of ATF4 and may be involved in protection mechanism in preimplantation embryos from oxidative injury.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2004.07a
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• pp.31-31
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• 2004