• Title/Summary/Keyword: Developing embryo

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Effects of Electric Stimulation and Activation Conditions on the Fusion and Development of Porcine Somatic Cell Nuclear Transfer Embryos (전기적 융합과 활성화 방법이 돼지 체세포 복제수정란의 체외발달에 미치는 영향)

  • 정기화
    • Journal of Embryo Transfer
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    • v.19 no.1
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    • pp.43-51
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    • 2004
  • The present study was conducted to investigate the effects of fusion and/or activation protocol on in vitro development of porcine somatic cell nuclear transfer (SCNT) embryos. Porcine fetal fibroblast cells were transferred into the perivitelline space of enucleated in vitro matured oocytes. Cell fusion and activation were induced simultaneous fusion/activation (SA) or delayed activation (DA) with or without cytochalasin B (CB) treatment with electic pulses in 0.28 M mannitol-based medium. The SCNT embryos were cultured in vitro for 7 days and stained with Hoechst 33342 to determine the number of nuclei. After 7 days culture, cleavage and blastocyst formation rates were 72.4% and 7.6% in SCNT and 76.3% and 20.4% in parthenotes. To examine the effect of electric field strengths on development of SCNT embryos, oocytes were fused two pulses of 110 V/mm, 130 V/mm or 150 V/mm for 30 sec post-injection. The fusion and cleavage rates in 130 V/mm group (70.2% and 72.6%) and 150 V/mm group (72.6% and 70.5%) were higher (P<0.05) than 110 V/mm group (47.1% and 48.6%), respectively. However, the rate of embryos developing to the blastocyst stage (8.1%, 9.7% and 10.7%) were not different among three groups. The cleavage rates and the blastcyst formation rates were not different among three treatment groups (SA group, 71.4% and 9.7%; SA+CB treatment group, 74.7% and 8.0%; DA+CB treatment group, 70.8% and 11.2%, respectively). And, no different in the number of cells in blastocysts was observed among the three groups (22.5$\pm$12.8, 23.3$\pm$11.2 and 21.6$\pm$10.4, respectively). These result suggest that two pulses of 130 V/mm or 150 V/mm for 30 sec with SA treatment or DA treatment are enough for fusion/activation of porcine somatic cell nuclear transfer (SCNT) embryos to develop to the blastocyst stage.

Egg Antibody Farming and IgY Technology for Food and Biomedical Applications (식품과 생의학을 위한 계란 항체생산과 IgY 기술의 활용)

  • Sim, Jeong S.;Sunwoo, Hoon H.
    • Proceedings of the Korea Society of Poultry Science Conference
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    • 2003.07b
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    • pp.37-54
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    • 2003
  • It has been recognized that the hen. like its mammalian counterparts. provides young chicks with antibodies as protection against hostile invaders. This system facilitates the transfer of specific antibodies from serum to egg yolk. and provides a supply of antibodies called immunoglobulin Y(IgY) to the developing embryo and the hatched chick. The protection against pathogens that the relatively immuno-incompetent newly hatched chick has. is through transmission of antibodies from the mother via the egg. Egg yolk. therefore. can be loaded with a large amount of IgY against pathogens which can immobilize the existing or invading pathogens during the embryo development or in day-old chicks. Thus. the immunization of laying hens to various pathogens results in production of different antigen-specific IgY in eggs. Egg yolk contains 8~20 mg of immunoglobulins (IgY) per $m\ell$ or 136~340 mg per yolk suggesting that more than 30 g of IgY can be obtained from one immunized hen in a year. By immunizing laying hens with antigens and collecting IgY from egg yolk. low cost antibodies at less than $10 per g compared to more than $20.000 per g of mammalian IgG can be obtained. This IgY technology opens new potential market applications in medicine. public health veterinary medicine and food safety. A broader use of IgY technology could be applied as biological or diagnostic tool. nut-raceutical or functional food development. oral-supplementation for prophylaxis. and as pathogen-specific antimicrobial agents for infectious disease control. This paper has emphasized that when IgY-loaded chicken eggs are produced and consumed. the specific antibody binds. immobilizes and consequently reduces or inhibits the growth or colony forming abilities of microbial pathogens. This concept could serve as an alternative agent to replace the use of antibiotics. since today. more and more antibiotics are less effective in the treatment of infections. due to the emergence of drug-resistant bacteria.

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Establishment of the expression system of human HtrA2 in the zebrafish (Zebrafish 동물모델에서 human HtrA2의 expression system 정립에 관한 연구)

  • Cho, Sung-Won;Park, Hyo-Jin;Kim, Goo-Young;Nam, Min-Kyung;Kim, Ho-Young;Ko, In-Ho;Kim, Cheol-Hee;Rhim, Hyang-Shuk
    • Journal of Life Science
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    • v.16 no.4
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    • pp.571-578
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    • 2006
  • HtrA2/Omi, a mitochondrial trypsin-like serine protease, is pivotal in regulating apoptotic cell death. Several lines of recent evidence suggest that HtrA2 is associated with the pathogenesis of neurodegenerative disorders; however, the physiological function of HtrA2 still remains elusive. For studying physiological function of HtrA2 in depth, it is necessary to develop a suitable expression system in the model animal. We therefore utilized the zebrafish as a model animal to establish expression of human HtrA2 (hHtrA2) in vivo. For expression of mature HtrA2 as GFP fusion in zebrafish embryos, the HtrA2 (WT) or (S306A) cDNAs with the C-terminal GFP tag were inserted into the pCS2+ plasmid. Expression patterns of HtrA2 in HEK293 cells were first monitored by immunofluorescence staining and immunoblot assays, showing approximately 64 kDa of the HtrA2-GFP fusion proteins. Subsequently, the hHtrA2 plasmid DNA or in vitro transcribed mRNA was microinjected into zebrafish embryos. The expression patterns of HtrA2 in Zebrafish embryos were monitored by GFP fluorescence in 24 hours-post-fertilization (hpf). Although expression patterns of HtrA2-GFP in developing embryos were different between the injected DNA and mRNA, both nucleic acids revealed good expression levels to further study the physiological role of HtrA2 in vivo. This study provides a suitable condition for expressing hHtrA2 in the zebrafish embryos as well as a method for generating useful system to investigate physiological properties of the specific human genes.

The theory and application of holistic innovation policy: Cases of Finland and Korea (총체적 혁신정책의 이론과 적용: 핀란드와 한국의 사례)

  • Seong, Ji-Eun;Song, Wi-Chin
    • Journal of Korea Technology Innovation Society
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    • v.10 no.3
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    • pp.555-579
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    • 2007
  • This study analysed theory and application of holistic innovation policy in the 3rd generation innovation policy with cases of Finland and Korea. Innovation policy areas of both countries are expanding from simple goal of economic development to extensive goals such as sustainable development, quality of life, balanced growth etc. Also administrative system changed in order to let technical innovation on the center of national operation and reorganized relation and structure of relative policies like manpower, education, region policy. Particularly, Korea is in embryo standing for S&T driven society since Noh's government. But, there are differences in embodiment degree and feature between both countries. Finland reinvigorate field of public opinion due to corporatism tradition as a Scandinavian institution. Government support system developed co-program and co-policy beyond territory of individual ministry to raise policy effectiveness. However, concept of the Korean innovation policy is ahead of the times but maintained framework of developing country. And there is no active participation of private sector and civil society because transition process accomplished on the top-down method, It could hardly progress toward transition in spite of various efforts to carry out a reform.

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Spawning Performance, Embryonic Development and Early Viability under Different Salinity Conditions in a Euryhaline Medaka Species, Oryzias dancena (서로 다른 염분도 조건하에서 광염성 송사리 Oryzias dancena의 산란, 발생 및 초기 생존)

  • Cho, Young-Sun;Lee, Sang-Yoon;Kim, Dong-Soo;Nam, Yoon-Kwon
    • Korean Journal of Ichthyology
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    • v.22 no.1
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    • pp.25-33
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    • 2010
  • Effects of different salinity levels on spawning performance, embryonic development and early viability of a euryhaline medaka species, Oryzias dancena, were examined. O. dancena were able to spawn eggs in a wide range of salinity from 0 to 70$^{\circ}/_{\circ\circ}$, however, the spawning frequency was lowered in complete freshwater (0$^{\circ}/_{\circ\circ}$) and in highly salted water (70$^{\circ}/_{\circ\circ}$). Fertilization success was negatively affected when the environmental salinity was higher than the salt concentration found in normal seawater. Embryonic viability and hatching success were also inversely related with the salinity levels. Typical abnormality was observed in developing embryos incubated at high salinities (30, 45 and 60$^{\circ}/_{\circ\circ}$). In addition, the time to hatch was significantly delayed with increasing salinities: peak hatching occurred at 12~14 days post fertilization (dpf) in freshwater and at least at 17 to 18 dpf in 60$^{\circ}/_{\circ\circ}$. Mean survival rates of the hatched larvae up to 7 days post hatching (dph) were at least 97% in salinity levels ranging from 0 to 30$^{\circ}/_{\circ\circ}$. However, larvae reared in 45 and 60$^{\circ}/_{\circ\circ}$ experienced significant mortality, especially in the early phase, resulting in only 75% and 64% survival rates up to 7 dph, respectively.

Comparison of teratogenecity induced by nano- and micro-sized particles of zinc oxide in cultured mouse embryos

  • Jung, A Young;Jung, Ki Youn;Lin, Chunmei;Yon, Jung-Min;Lee, Jong Geol;Lee, Beom Jun;Yun, Young Won;Nam, Sang-Yoon
    • Korean Journal of Veterinary Research
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    • v.55 no.2
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    • pp.133-139
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    • 2015
  • The increasing uses of zinc oxide nanoparticles (nZnO) in industrial and personal care products raise possible danger of using nZnO in human. To determine whether ZnO induces size-dependent anomalies during embryonic organogenesis, mouse embryos on embryonic day 8.5 were cultured for 2 days under 50, 100, and $150{\mu}g$ of nZnO (< 100 nm) or micro-sized ZnO (mZnO; $80{\pm}25{\mu}m$), after which the morphological changes, cumulative quantity of Zn particles, and expressions of antioxidant and apoptotic genes were investigated. Although embryos exposed to $50{\mu}g$ of ZnO exhibited no defects on organogenesis, embryos exposed to over $100{\mu}g$ of ZnO showed increasing anomalies. Embryos treated with $150{\mu}g$ of nZnO revealed significant changes in Zn absorption level and morphological parameters including yolk sac diameter, head length, flexion, hindbrain, forebrain, branchial bars, maxillary process, mandibular process, forelimb, and total score compared to the same dose of mZnO-treated embryos. Furthermore, CuZn-superoxide dismutase, cytoplasmic glutathione peroxidase (GPx) and phospholipid hydroperoxidase GPx mRNA levels were significantly decreased, but caspase-3 mRNA level was greatly increased in nZnO-treated embryos as compared to normal control embryos. These findings indicate that nZnO has severer teratogenic effects than mZnO in developing embryos.

Spawning Behavior and Development of Eggs and Larvae of the Korea Freshwater Goby, Rhinogobius brunneus (Gobiidae: Perciformes) (한국산 밀어 (Rhinogobius brunneus)의 산란습성, 난발생 및 자어의 형태발달)

  • HAN Kyeong-Ho;KIM Yong Uk;CHOE Kyu-Jung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.31 no.1
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    • pp.114-120
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    • 1998
  • The spawning behavior, development of eggs and larvae of the Korea freshwater goby, Rhinogobius brunneus (Temminck et Schlegel) were studied. The eggs were spawned as a one-layer mass, hanging from the underside of a small pebble, and guarded by one male. The eggs were elliptic, about 1.48 mm in length and 0.65 mm in breadth, with a round top and a somewhat flat base with glutinous fibers. Hatching in the indoor tank with $17.0^{\circ}C$ in mean water temperature started from the 146 hours after fertilization. In the late developing stages, the embryo moved and freely changed its head to face the free tip of the chorion (normal egg) or to the basal end with a boundle of adhesive filaments (agrippa egg). Newly-hatched larvae were $3.10\~3.30$ mm in total length (mean: 3.22 mm), and mouth and anus were not yet open. Melanophores were present on the air- bladder, around the anus, and on the ventral part of the caudal region. The larva $3\~4$ days old transformed to postlarval stage, and they were $3.30\~3.85$ mm in total length (mean: 3.60 mm). As yolk sac and oil globules werw nearly absorbed, mouth and anus were open, and they fed rotifers actively. In $20\~22$ days after hatching the larvae grew to 5.85 mm in 71, and the caudal notochord flex at $45^{\circ}$.

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TEMPOROSPATIAL PATTERNS OF PROGRAMMED CELL DEATH DURING EARLY DEVELOPMENT OF THE MOUSE EMBRYOS (생쥐 배자발생초기의 세포자기사 발현 양상에 관한 연구)

  • Baik, Byeong-Ju;Lee, Seung-Ik;Kim, Jae-Gon;Park, Byung-Yong;Park, Byung-Keon
    • Journal of the korean academy of Pediatric Dentistry
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    • v.28 no.4
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    • pp.709-727
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    • 2001
  • The pattern of programmed cell death(PCD) has been examined during the early developmental period of development in mouse embryos, from embryonic day 4.5(E4.5) to E11.5 Embryos from Balb/c breedings were harvested at various embryonic stages between E4.5 and El1.5. Cell death was analysed by in situ terminal deoxynucleotidyl transferase mediated dUTP nick end labeling(TUNEL) staining in tissue sections and whole embryos. At the blastocyst stage(E4.5), a very few apoptotic cells were found in the inner cell mass of the blastocyst. In the early egg cylinder stage(35.0-5.5), a few apoptotic cells were detected in the embryonic ectoderm, the embryonic endoerm and the proamniotic cavity. In the advanced egg cylinder stage(E5.5-6.5), TUNEL-posifive cells were observed in the extra-embryonic ectoderm and extra-embryonic endoderm as well as in the embryonic ectoderm, embryonic visceral endoderm and proamniotic cavity. In the streak stage(E6.75-7.75), many TUNEL-positive cells were found in the ectoplacental cone. In contrast, only very few apoptotic cells were found in the chorion and extra-embryonic endoderm in extra-embryonic regions. In intra-embryonic region, a few apoptotic cells were randomly found in the embryonic ectoderm, mesoderm and visceral endoderm. At the early somitogenesis stage(E8.0-8.5), most apoptotic cells were observed in the most cranial portion of neural fold (neural ectoderm and adjacent ectoderm). At the mid somitogenesis stage(39.0-9.5), the otic placode first showed TUNEL-positive at this stage. Small number of TUNEL-positive cells were also first seen around optic placode and branchial arches. Three streams of TUNEL-positive cells were clearly seen in the cranial region at 59.5-9.75. At E10.5, apoptotic cells were localized in the developing eye, the junctional portion of medial nasal, lateral nasal and maxillary processes, the lateral portion of branchial arches, the junction of bilateral mandibular processes, and apical ectodermal ridges of limb buds. At E11.5, apoptotic cells were noticeably decreased in most area, except the developing limbs and several somites in the tail region. In this study, the global temporospatial pattern of PCD throughout early development of mouse embryos was discussed. It may provide the basis for further studies on its role in the morphogenesis of the embryo.

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