• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2004년도 춘계학술발표대회
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• pp.283-283
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• 2004

The eggs of most animal species are surrounded by an extracellular matrix known as chorion, egg envelope, egg coat, or zona pellucida. Development of fish embryo usually takes several days in an aquatic environment. During embryonic development, the chorion must protect embryo from physical damage and microbial infection in the exposed aquatic environment. (omitted)

• BMB Reports
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• 제40권1호
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• pp.22-28
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• 2007

MyoD, expressed in skeletal muscle lineages of vertebrate embryo, is one of muscle-specific basic helix-loop-helix (bHLH) transcription factors, which plays a key role in the determination and differentiation of all skeletal muscle lineages. In this study, a cDNA of grass carp MyoD was cloned and characterized from total RNA of grass carp embryos by RT-PCR. The full-length cDNA of grass carp MyoD is 1597 bp. The cDNA sequence analysis reveals an open reading frame of 825 bp coding for a protein of 275 amino acids, which includes a bHLH domain composed of basic domain (1-84th amino acids) and HLH domain (98-142th amino acids), without signal peptide. Then the MyoD cDNA of grass carp was cloned to yeast expression vector pPICZ$\alpha$A and transformed into P. pastoris GS115 strain, the recombinant MyoD protein with a molecular weight of about 31KD was obtained after inducing for 2d with 0.5% methanol in pH 8.0 BMGY medium, and the maximum yield was about 250 mg/L in shaking-flask fermentation. The results were expected to benefit for further studies on the crystal structure and physiological function of fish MyoD.

• Clinical and Experimental Reproductive Medicine
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• 제36권2호
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• pp.101-109
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• 2009

목 적: 소 수정란의 telomeric DNA 함량 및 텔로머레이스 활성도 (telomerase activity)를 분석하여 초기 배아형성과정에서 텔로미어의 변화 양상을 규명하고자 하였다. 연구방법: 소의 초기 배 발생단계별 체외수정란을 대상으로 양적형광보인법 (Q-FISH)을 이용한 telomeric DNA의 함량 분석과 Telomeric Repeat Amplification Protocol (TRAP)로서 텔로머레이스 활성도를 분석하였다. 결 과: 소의 초기 수정란의 telomeric DNA 함유율은 발생과정이 진행됨에 따라 유의적인 양적 증가를 나타내었다. 배반포배의 질 (quality)에 따른 텔로미어의 함량 차이는 나타나지 않았다. 소 수정란의 8 세포기, 상실배기 및 배반포기의 모든 초기 배 단계에서 텔로머레이스 활성도를 보였으며, 상실배에서 증가하여 배반포배에서 가장 강한 활성도를 나타내었다. 결 론: 소의 초기 배아형성과정이 진행되면서 텔로미어의 함량 및 텔로머레이스 활성도는 점진적으로 증가하고 이들 간에는 정 (positive)의 상관관계를 가진다. Telomeric DNA 함량의 증가를 유지하기 위하여 텔로머레이스의 활성화는 필수적인 것으로 보여진다. 배반포기 배아에서 높은 텔로미어 함량과 강한 텔로머레이스 활성도는 이 시기가 배발생 및 분화과정 중 가장 세포적 활성도가 높은 시점임을 의미하는 것으로 사료된다.

• 대한생식의학회:학술대회논문집
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• 대한불임학회 2005년도 제49차 추계학술대회
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• pp.107-108
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• 2005

• 한국수산과학회지
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• 제29권2호
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• pp.157-164
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• 1996

태생 경골어류 망상어, Ditrema temmincki의 임신기 동안 체내자어는 건중량과 체액삼투압의 증가를 나타냈으며, 모체는 난소강액의 증가와 함께 난소 내부조직상의 현저한 변화를 나타냈다. 체내자어는 난황흡수 완료시기인 전장 6.0 mm부터 전장 60.0 mm 사이에 373.76mg의 건중량 증가를 나타냈으며, 체액삼투압은 출산직전인 전장 64.0mm에는 796.8mOsmo1/kg으로 나타났다. 모체의 난소외막은 수정 이후 뚜렷히 비후되기 시작하여 임신중기인 3월에 가장 비후되고, 난소습곡은 임신후기인 4월말에 가장 팽창되며, 혈관의 팽창 및 혈구의 증가는 출산직전에 최고에 달한다. 이와같은 변화양상은 임신기 동안 체내자어와 모체의 영양${\cdot}$환경적 적응현상을 의미하는 것이라 하겠다.

• 한국가축번식학회지
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• 제26권4호
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• pp.385-394
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• 2002

This study was constructed the correlations of the embryonic developmental rates and the frequency of chromosome aberration using ear-skin-fibroblast cell in nuclear transfer (NT) derived embryos. Karyoplast-oocyte complexes were fused and activated simultaneously, then cultured for seven days to assess development. The developmental rates of NT and in vitro fertilization (IVF) embryos were 55.4% vs 63.5%, 31.7% vs 33% and 13.4% vs 16.8% in 2 cell, 8 cell and blastocyst, respectively. Firstly, the frequency of chromosome aberrations were evaluated using fluorescent in situ hybridization (FISH) technique with porcine chromosome 1 submetacentric specific probe. Chromosome aberration was detected at day 3 on the embryo culture, the percentages of chromosomal aneuploidy in NT and IVF embryos at 4-cell stage were 40%, 31.3%, respectively. Secondly, embryonic fragmentation was evaluated at 4-cell stage embryo. Frequency of embryonic fragmentations was in 51.3% of NT, 61.3% of IVF, 28.9% of parthenogenetic activation at 4-cell stage. The proportion of fragmentation in NT embryos was higher than activation embryos. This result indicates that chromosomal abnormalities and embryonic fragments are associated with low developmental rate in porcine NT embryo. It is also suggest that abnormal porcine embryos produced by NT related with lower implantation rate, increased abortion rate and production of abnormal fetuses.

• 한국양식학회지
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• 제12권3호
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• pp.197-204
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• 1999

Successful produciton of seedlings for marine fish species are highly influenced by the aquisiton of sufficient number of good-quality eggs on required time. human chorionic gonadotropin has been used to induce successful ovulation for the purpose. Rabbitfish, siganus canaliculatus, is inhabited along the coast of Cheju Province and treated valuably for raw fish and fish roasted with seasoning. Female wild-captured rabbitfish (314 to 279 g in body weigth) were injected into intraperitoneal space with human chorionic gonadotropin (HCG) of 5,00,1,000, and 1,500 IU/kg body weight during spawning season of July 14 (trial 1), and July 27 (trial 2). Test fish in trial 1 were injected two times, first on July 14 and second on July 21 with one weekinterval because of no ovulation symptoms, but just one time in trial 2. All females injected both 1,000 and 1,500 IU/kg body weight ovulated in all trials. Among 500 IU/kg injection treatments, just one female ovulated in trial 1 showing 25% of ovulation rate, but none ovulated in trial 2. Ovulation was not occurred in control goups during test period. Time to reach ovulation after injection was 172 to 270 hours in trial 1 and 77 to 132 hours in trial 2, showing shorter as spawning season approached. Generally, ovulation of smaller females were late than that of larger ones. Gonadosomatic index (GSI) and pseudo-gonadosomatic index (Pseudo-GSI) were ranged 22 to 33% and 19 to 28%, respectively. One female spawned 406,200 to 1,032,000 eggs, and obtainable eggs per 100g of body weight were calculated by 130,000 to 190,000. Spawning rates were comparatively high by 96.0 to 98.4%. Rates to reach embryo-formation and hatching were higher in 500 high by 96.0 to 98.4%. Rates to reach embryo-formation and hatching were higher in 500 and 1,000 IU/kg treatments than in 1,500 IU/kg treatment. The result of present study demontrates that HCG treatment during spawning season could become very useful for ovulation of rabbitfish and the suitable dosage was suggested as 1,000 IU/kg of body weight.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.101-101
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• 2002

GH-transgenic coho salmon (Oncorhynchus kitsutch) juveniles in tGH＊T$_3$and tGH*PTU were fed with the diets containing 1 ug/g fish of 3,5,3'-triiodo-L-thyronine (T$_3$) and 30 ug/g fish of 6-n-propyl-2- thiouracil (PTU), respectively, to assess the effect of these drugs on the change of physiological activity, growth and survival rate in comparison with normal transgenic (tGH＊C) and nontransgenic coho salmon (Wild) for 90 days. Although the daily food intakes of all transgenic (tGH)-groups were higher than Wild, the amount was reduced by exogenous PTU supply. The fred efficiencies of tGH-groups were lower than Wild, but the efficiency was reduced both by T$_3$and PTU. The survival rate of tGH-group was significantly higher than that of Wild, but there was no significant difference among tGH-groups. Although the growth of tGH-coho salmon was faster than Wild. the growth rate of transgenic salmon was increased by exogenous T$_3$, but was reduced by PTU Plasma TT$_4$levels of tGH-groups was approximately 2-fold higher relative to Wild, but there were no difference of plasma TT$_4$levels among tGH-groups. plasma TT$_3$level or tGH-coho salmon was increased by exogenous T$_3$administration, but was reduced by exogenous PTU. In addition, although plasma GH levels of all tGH-groups were higher than that of Wild, the GH level in plasma of transgenic coho salmon was increased by exogenous T$_3$and reduced by exogenous PTU. In the meantime, the transgenic fishes also displayed head, jaw and opercular abnormalities typical of the offsets of this gene construct in coho salmon, indicating that some imbalance in growth processes has been induced. However, the abnormalities of transgenic coho salmon was reduced following exogenous PTU administration.

• Clinical and Experimental Reproductive Medicine
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• 제47권4호
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• pp.293-299
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• 2020

Objective: The goal of the present study was to investigate the rate of chromosomal aneuploidies in surplus embryos after sex determination at the cleavage stage. Then, the same chromosomal aneuploidies were evaluated in blastocysts after extended culture. Methods: Sixty-eight surplus embryos were biopsied at the cleavage stage and incubated for an additional 3 days to allow them to reach the blastocyst stage. The embryos were reanalyzed via fluorescence in situ hybridization (FISH) to examine eight chromosomes (13, 15, 16, 18, 21, 22, X, and Y) in both cleavage-stage embryos and blastocysts. Results: Although the total abnormality rate was lower in blastocysts (32.35%) than in cleavage-stage embryos (45.58%), the difference was not significant (p=0.113). However, when we restricted the analysis to autosomal abnormalities, we observed a significant difference in the abnormality rate between the cleavage-stage embryos (44.11%) and the blastocysts (17.64%, p=0.008). A higher rate of sex chromosomal abnormalities was also observed in cleavage-stage embryos (29.4%) than in blastocysts (14.70%, p=0.038). Conclusion: The data indicated that embryo biopsy should be conducted at the blastocyst stage rather than the cleavage stage. The results also emphasized that examination of common chromosomal aneuploidies apart from sex selection cycles can be conducted in the blastocyst stage with the FISH method.

• 한국가축번식학회지
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• 제20권3호
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• pp.323-331
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• 1996

포유동물에 있어서 조기 성 판정기술은 축산에 있어서의 성별 육종프로그램이나 인간의 X-염색체 관련 열성유전병의 산전진단 등 여러 분야에 응용될 수 있다. 초기배에 대한 성 판정은 성염색체에 존재하는 특이한 염기서열을 증폭시키는 polymerase chain reaction (PCR)과 X와 Y 염색체에 대한 특이적 probe를 이용하는 fluorescent in situ hybridization (FISH)에 의하여 수행될 수 있다. 1992년과 93년, 2개년도에 걸쳐 본 연구실에서 돼지의 3.3 kb 웅성특이 DNA 절편(pEM39)을 cloning하였다. 본 연구는 pEM39가 성특이 DNA-probe로 이용될 수 있는지를 조사하기 위해 PCR과 FISH를 이용하였다. 돼지 난자는 도축장에서 구입한 돼지 난소로부터 채취되었고, 체외배양후 체외수정되었다. 한편 처녀발생나자를 negative control로 이용하였다. 2 세포기의 수정란을 선발한 후 PCR을 통하여 DNA를 분석한 결과, 10개의 수정란 중 6개는 자성, 다른 4개는 웅성으로 판정되었으며, FISH를 수행한 결과, done된 웅성특이 DNA 단편은 돼지 간조직과 초기배에서 웅성특이성을 보였다. 또한 FISH와 karyotyping을 수행한 결과 clone된 웅성특이 DNA 단편이 Y 염색체 q-arm의 heterochromatic region에 위치함을 알 수 있었다. 이러한 결과로 보아 clone된 웅성특이 DNA 단편이 초기배의 성을 조기판정하는데 있어 유용하리라 사료되며, PCR에 의한 초기배의 성 판정에 있어 신뢰할만할 지표가 될 수 있을 것이다.