• Title/Summary/Keyword: 배아 발생

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Effect of Cobalt (II) on the Fertilization and Embryo Development of the Sea Urchin ($Hemicentrotus$ $pulcherrimus$) (코발트(II)가 말똥성게($Hemicentrotus$ $pulcherrimus$)의 수정 및 배아 발생에 미치는 영향)

  • Hwang, Un-Ki;Ryu, Hyang-Mi;Choi, Yong-Hwan;Lee, Seung-Min;Kang, Han-Seung
    • Korean Journal of Environmental Biology
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    • v.29 no.4
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    • pp.251-257
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    • 2011
  • Cobalt is a naturally occurring element found in rocks, soil, water and/or is among the harmful pollutants as generated by industrialized. In the environment, cobalt has two oxidation states, cobalt (II) (Co (II)) and cobalt (III) (Co (III)). If coastal water is contaminated by cobalt, it through the food chain can have an impact on marine ecosystems. Therefore, we examined the gametotoxic and embryotoxic effects of Co (II) at various concentrations (10, 100, 500, 1000, 2500 ppb) in the sea urchin $Hemicentrotus$ $pulcherrimus$. Spawning was induced by injecting 1 mL of 0.5 M KCl into coelomic cavity. Males released white or cream-colored sperms and females released yellow or orange-colored eggs. Experiment was begun within 30 min the collection of both gametes. The fertilization and embryo development rates test were performed for 10 min and 64 h after fertilization, respectively. The fertilization rates in the control condition (not including Co (II)) and experimental group were not significantly changed. The embryo development rates in the control condition were greater than 90% and were significantly decreased with concentration dependent manner. The normal embryogenesis rate was significantly inhibited in exposed to cobalt (II) ($EC_{50}$=71.84 ppb, 95% Cl=16.71-203.36 ppb). The NOEC and LOEC of normal embryogenesis rate were <10 ppb and 10 ppb, respectively. These results suggest that the early embryo stages of $H.$ $pulcherrimus$ have toxic effect at greater than 10 ppb of Co (II) concentration.

Effect of Fragment Removal on Development of Human Fragmented Embryos in IVF-ET Program (IVF-ET 시술 시 파편제거가 파편화된 인간배아의 발달에 미치는 효과)

  • Chi, Hee-Jun;Koo, Jung-Jin;Lee, Ju-Ok;Ryu, Hyoung-Eun;Kim, Kwang-Rae;Park, Chan;Roh, Sung-Il
    • Clinical and Experimental Reproductive Medicine
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    • v.37 no.4
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    • pp.339-348
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    • 2010
  • Objective: To investigate the beneficial effect of fragment removal on the subsequent cell division and clinical outcome of the fragmented human embryos. Methods: A prospective study was performed in Hanna Women's Clinic and Mizmedi Hospital. Sixty couples undergoing In vitro fertilization-embryo transfer (IVF-ET) program were participated in the present study. The microsurgical fragment removal was performed in 106 fragmented embryos of 29 patients before the transfer. As a control group, 122 fragmented embryos of 31 patients were transferred without the fragment removal. Effects of fragment removal on morphological changes and clinical outcomes of fragmented embryos were investigated. Results: Mean morphological grade (G2.79) of fragmented embryos was significantly improved after the fragment removal(G1.63, p<0.001). Most of the fragmented embryos did not show a regeneration of fragments after the fragment removal during the subsequent development, and a beneficial effect of fragment removal on the development of the fragment removed embryos was observed. Implantation and pregnancy rates of fragment removed embryos were 12.3% and 31.3%, whereas the rates of control group embryos were 6.6% and 22.5%, respectively. There was no statistical significance in the rates between the two groups because of the low number of trials. Conclusion: Microsurgical fragment removal improved the subsequent development as well as the morphological grade of fragmented embryos. The fragment removal may be beneficial for neighboring blastomeres by repairing the intercellular communication and removing the secretion of the potential toxic materials by fragments.

Evaluation of Water Quality Using Fire-bellied Toad (Bombina orientalis) Embryos in Tancheon Basin (무당개구리 배아를 이용한 탄천 수계 수질에 대한 생물학적 평가)

  • Park, Chan Jin;Song, Sang Ha;Kim, Dae Han;Gye, Myung Chan
    • Korean Journal of Environmental Biology
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    • v.33 no.4
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    • pp.425-432
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    • 2015
  • Pollution in the fresh water system in urban area has the adverse effect on the amphibians population. Restoration activity of amphibian in the urban stream has been growing in Korea as well as western country. For successful restoration water quality of urban stream should be sufficient for survival and normal development of amphibian. To monitor the biological safety of surface water in the Tancheon basin, the capital area of Korea, a 6-day exposure Bombina orientalis embryo developmental toxicity assay was examined. The toxicity of surface water of Tancheon mainstream were lower than those of tributaries of Tancheon. The survival rate of embryos negatively correlated with total dissolved solid, turbidity and electrical conductivity whereas the developmental abnormality and growth retardation of embryos was positively correlated with total dissolved solid, turbidity and electrical conductivity. An amphibian developmental toxicity assay would be helpful for the selection of point for construction of habitat and reintroduction of amphibian in interrupted urban stream.

Effects of Nitric Oxide Donor Supplementation on Copper Deficient Embryos and Nitric Oxide-Mediated Downstream Signaling (Nitric Oxide Donor 첨가가 구리 결핍 배아의 발달과 Nitric Oxide 하위 신호전달체계에 미치는 영향)

  • Yang, Soo-Jin
    • Journal of Nutrition and Health
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    • v.41 no.8
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    • pp.691-700
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    • 2008
  • One suggested mechanism underlying copper (Cu) deficiency teratogenicity is a low availability of nitric oxide (NO), signaling molecule which is essential in developmental processes. Increased superoxide anions secondary to decreased activities of Cu-zinc superoxide dismutase (Cu-Zn SOD) in Cu deficiency can interact with NO to form peroxynitrite, which can nitrate proteins at tyrosine residues. In addition, peroxynitrite formation can limit NO bioavailability. We previously reported low NO availability and increased protein nitration in Cu deficient (Cu-) embryos. In the current study, we tested whether Cu deficiency alters downstream signaling of NO by assessing cyclic GMP (cGMP) and phosphorylated vasodilator-stimulating phosphoprotein (VASP) levels, and whether NO supplementation can affect these targets as well as protein nitration. Gestation day 8.5 embryos from Cu adequate (Cu+) or Cu- dams were collected and cultured in either Cu+ or Cu- media for 48 hr. A subset of embryos was cultured in Cu- media supplemented with a NO donor (DETA/NONOate; 20 ${\mu}M$) and/or Cu-Zn SOD. Cu-/Cu- embryos showed a higher incidence of embryonic and yolk sac abnormalities, low NO availability, blunted dose-response in NO concentrations to increasing doses of acetylcholine, low mRNA expression of endothelial nitric oxide synthase (eNOS), increased levels of 3-nitrotyrosine (3-NT) compared to Cu+/Cu+ controls. cGMP concentrations tended to be low in Cu-/Cu- embryos, and they were significantly lower in Cu-/Cu- yolk sacs than in controls. Levels of phosphorylated VASP at serine 239 (P-VASP) were similar in all groups. NO donor supplementation to the Cu- media ameliorated embryonic and yolk sac abnormalities, and resulted in increased levels of cGMP without altering levels of P-VASP and 3-NT. Taken together, these data support the concept that Cu deficiency limits NO availability and alters NO/cGMP-dependent signaling in Cu- embryos and yolk sacs, which contributes to Cu deficiency-induced abnormal development.

Serological detection of barley stripe mosaic virus infection in the seeds of barley and wheat cultivars grown in Korea (혈청학적방법에 의한 보리와 밀종자의 보리${\cdot}$줄무늬 모자이크 바이러스 감염상조사)

  • La Yong-Joon;Park Yang-Kyo
    • Korean journal of applied entomology
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    • v.18 no.1 s.38
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    • pp.29-33
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    • 1979
  • An attempt was made to find out whether barley stripe mosaic virus (BSMV) occurs in barley and wheat cultivars in Korea. Modified Hamilton's technique for serological detection of barley stripe mosaic virus was used to assay for BSMV in mature seed embryos. Of the 51 barley and wheat seed lots assayed, BSMV was detected in 34 of them. BSMV was detected in covered, naked, and matting barley as well as in wheat. The level of BSMV infection in the infected cultivars varied from $2\~35\%$. This is the first reported occurrence of barley stripe mosaic virus in Korea.

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Comparison of Cell Numbers in Inner Cell Mass vs Trophectoderm and Establishment of ES-like Cells Derived from Day 7 to 9 IVF Bovine Blastocysts (소 체외수정 유래 7~9일령 배반포내에서 내세포괴 대영양배엽세포의 비율 및 배아간세포 확립율의 비교)

  • 공일근
    • Korean Journal of Animal Reproduction
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    • v.21 no.2
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    • pp.157-165
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    • 1997
  • 포유류 배반포배의 epiblast는 내세포괴에 포함되어 있으며, 이 epiblast cells이 배 및 태아의 생식세포와 일반 체세포로 분화된다 (Beddington, 1986; Lawson 등, 1991). 그런데 조기에 발달된 부화배반포기 배가 지연발생된 부화배반포기 배보다도 많은 epiblast cells을 가지고 있다고 한다(Talbot 등, 1995). 그래서 본 연구에서는 체외수정 유래의 배반포배의 발육속도에 따른 내세포괴/영양배엽세포의 비율 및 배아간세포 확립 효율을 비교하여 발달일령 간에 차이가 있는지를 규명하고자 하였다. 공시한 소의 난포란을 TCM-199에 0.5$\mu\textrm{g}$/ml FSH, 5$\mu\textrm{g}$/ml LH, 10% FBS, 100 units/ml penicilin, 및 100$\mu\textrm{g}$/ml streptomycin을 첨가하여 39$^{\circ}C$, 5% CO2 조건하에서 24시간 동안 체외성숙한 후, 5$\mu\textrm{g}$/ml heparin으로 수정능이 획득된 1$\times$106 sperm/ml의 정자로 체외수정을 유도하였다. 체외수정 후 18~20시간에 과립막세포를 vortexing으로 제거하여 얻은 모든 체외수정란을 3mg/ml BSA, 20${\mu}\ell$/ml NEM amino acids, 40${\mu}\ell$/ml BME amino acids, 10mM glycine 및 1mM alanine이 함유된 CR1aa 배양액에서 BRL 단층세포와 공배양을 실시하였다. 수정후 7, 8 및 9일재 (체외수정일 : 0일)에 확장배반포기까지 발달한 수정란을 이중염색 및 배아간세포의 확립 실험에 공시하였다. 체외수정후 24시간에 분할된 총 1,145개의 수정란이 7, 8 및 9일째에 후기 배반포기까지 각각 222(15.6%), 103(7.2%) 및 52(3.6%)로 발달하여 총 377개 (26.4%)가 발달하였다. 내세포괴/영양배엽세포의 비율은 7일 및 8일째 배반포배에서 각각 47.2$\pm$11.9/95.1$\pm$24.4개 (33/67%) 및 40.3$\pm$12.4/83.3$\pm$26.9개 (33/67%)로서 9일째 배반포배의 19.3$\pm$8.1/62.3$\pm$23.1개 (24/76%) 보다 유의적(P<0.05)으로 높았다. ES-like cells을 확립하기 위하여 후기배반포기 배를 mouse embryonic fibroblast 단층 공배양기에 옮긴 후 5일에 내세포괴의 부착 여부를 판정하고, 10일에 배아간세포의 확립 여부를 판정하였다. 그 결과 7, 8 및 9일째의 배반포기배의 각각 47.7% (82/172), 30.9%(22/71) 및 15.6%(5/32)에서 배아간세포가 확립되었다. 이상의 결과에서 배반포기까지의 발육이 빠른 수정란에서 영양배엽에 대한 내세포괴세포의 비율이 높았고 배아간세포의 확립율도 높다는 사실을 입증할 수 있었으며, 이와 같은 결과에서 체외수정란 유래 배반포배의 질을 결정할 수 있을 것으로 생각된다.

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Genetic Studies on the Sea Urchin Embryogenesis and Skeletogenesis (성게의 발생과 뼈대형성의 유전학적 연구)

  • Lee, Youn-Ho
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.6 no.4
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    • pp.265-273
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    • 2001
  • The sea urchin has been used as sea food in many countries. This species has also been an important organism of embryological studies for more than a century. In recent years, sea urchin embryos are being used as testing materials for toxicity of pollutants and toxins. Usefulness of sea urchin embryos as experimental models comes from the easiness in obtaining sea urchin samples and a lot of gametes, in rearing embryos in the laboratory, in observing the cellular movement and organ formation during the embryogenesis and in manipulating blastomeres and genetic maferials. The sea urchin in itself is a key organism for the understanding of deuterostome evolution from the protostomes and of indirect development of marine invertebrates which undergo the planktotrophic larval stage. A fertilized sea urchin egg goes through rapid cleavage and becomes a 60 cell embryo 7hr after fertilization. It then develops into a morula, a blastula, a gastrula and finally a pluteus larva approximately 70 hr after fertilization. At the 60 cell stage, the embryo comprises of five territories that express territory-speciflc genes and later form different organs. Micromeres at the vegetal pole ingress into the blastoceol and become the primary mesenchyme cells(PMCs). PMCs express genes involved in skeletogenesis such as SM30, SM37, SM50, PM27, msp130. Among the genes, SM37 and SM50 are considered to be members of a gene family which is characterized by early blastula expression, Glycine-Proline-Glutamine rich repeat structures and spicule matrix forming basic proteins. Genetic studies on the sea urchin embryos help understand the molecular basis of indirect development of marine invertebrates and also of the biomineralization common to the animal kingdom.

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