• Clinical and Experimental Reproductive Medicine
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• 제27권4호
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• pp.345-351
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• 2000

Objective: Hatching of the blastocyst from the zona pellucida (ZP) is a key event in mammalian implantation. In vivo, two factors have been identified as possible mediators of hatching: lysis of the ZP by substances elaborated either from the embryo or female reproductive tract and pressure exerted on the zona by expansion of the blastocyst. Two methods of zona manipulation were already in use to enhance the ability of embryos to hatch: mechanical PZD and chemical ZD by acidic Tyrode's solution. But several controversies of each method have been reported. The purpose of this study was to investigate the effect of pronase for mouse embryo hatching. Methods: Mouse embryos were obtained following ovulation induction of $F_1$ animals. Fresh and cryo-thawed morula embryos were exposed to 0.5, 1.0, 2.0, 5.0 ${\mu}g/ml$ pronase in Ham's F10 for 72 hrs. Main outcome measures were the rates of partial hatching and completely hatched blastocysts, and cell number of it. Results: In fresh and cryo-thawed group, the rates of completely hatched blastocyst were significantly higher in 5 ${\mu}g/ml$ pronase treatment group than control group. There was no difference in completely hatched blastocyst total cell number between pronase treatment group and control group. This suggest that pronase treatment did not harmful in mouse embryo development. In pronase treatment group, zona pellucida were thinner than control group. Conclusion: The addition of pronase to culture media may accelerate the hatching of embryo. So, enzymatic treatment of the zona may provide a valuable and effective assisted hatching technique for human in-vitro fertilization-embryo transfer.

• 한국어류학회지
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• 제28권4호
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• pp.260-266
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• 2016

능성어, Epinephelus septemfasciatus의 우량 수정란을 생산할 수 있는 어미 선별기준을 설정하기 위하여 해상가두리에서 6년간 키운 시험어의 현태적 특징을 관찰하고 호르몬 처리하여 난질을 관찰 분석하였다. 시험어는 4개의 체중 범위(3.0~4.0, 4.0~5.0, 5.0~6.0, 6.0~7.0 kg)와 뒷지느러미를 기준으로 복부팽만한 정도를 I~IV 단계로 나눈 후 산란빈도, 채란량, 부상률, 수정률, 난발생 생존율 및 부화율을 조사하고, 수정란과 유구의 크기를 측정하였다. 또한 생식소 발달을 확인하기 위하여 복부팽만도 단계별 생식소 중량지수(GSI)를 구하고, 조직학적으로 관찰하였다. 그 결과 어체중이 5.0 kg 이상일 때 모든 개체에서 배란이 유도되었다. 또한 복부팽만도 I, II 단계에서는 배란이 유도되지 않고, III, IV 단계에서만 배란이 유도되었다. 난질은 III 단계가 우수하였고 그 때의 GSI는 $4.68{\pm}0.40%$였다.

• 한국어류학회지
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• 제20권4호
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• pp.255-262
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• 2008

멸종위기야생동식물 1급으로 지정된 꼬치동자개, Pseudobagrus brevicorpus의 종복원을 위해 인위적인 교배를 통하여 획득한 자치어를 대상으로 성분화 과정을 조사하여 성적임성 여부를 확인하였다. 부화 후 4~5일째 자어에서 생식융기 1쌍이 장과 중신관 사이에서 관찰되었고, 그 후 유사분열이 활발히 진행되었다. 부화 후 20일째 치어에서 난원세포의 출현으로 난소 분화가 시작되었으며, 부화 후 30~40일째를 거치면서 염색인기와 주변인기의 난모세포로 성장하였다. 난황은 부화 후 80일째의 자어에서 생성되기 시작하였고, 부화 후 100일째가 되어 eosin에 염색되는 난황구가 생성되었다. 이후 난모세포는 난황이 지속적으로 축적되면서 세포의 크기와 수가 증가하기 시작하였고, 부화 후 약 10개월째에는 난막과 여포세포층에 의해 둘러싸여진 난모세포로 분화하였다. 한편 정소는 부화 후 25일경 치어에서 분화되기 시작하였고, 40일경 유사분열을 통해 정원세포를 둘러싸는 포낭을 형성하였다. 부화 후 100일째가 되면 정소는 정소엽 구조를 형성하였고 hematoxylin에 매우 진하게 염색되는 정모세포로 분화하였다. 부화 후 7개월째 치어의 정소엽 내부에 정세포가 출현한 이후, 부화 후 만 1년을 경과한 치어에서 최초로 성숙한 정자가 출현하였다. 위 결과를 바탕으로 인공적으로 부화된 꼬치동자개 자치어의 성분화는 다른 종에서 보이는 것과 같이 정상적인 발달을 보인다고 판단되었다.

• 한국가축번식학회지
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• 제18권4호
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• pp.251-256
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• 1995

Two experiments were conducted to study the production of in vitro fertilized bovine embryos and the viability of blastocysts cryopreserved by vitrification. In experiment 1, production rate of in vitro matured bovine oocytes after fertilization in medium containing bovine oviduct epithelial cells (BOEC), cumulus cells and granulosa cells to blastocysts were 18.4, 14.6 and 13.1%, respectively. Developmental percentages of blastocysts produced at day 6, 7 and 8 were 8.5, 10.6 and 15.2% respectively. Hatching rate of bovine embryos produced was 60.0%. In experiment 2, post-thawed surviving embryos in a vitrification solution consisting of 7.15M ethylene glycol, 2.5 mM ficoll and 0.3 M sucrose were 36.4% (56/154). Also, survival rate of bovine embryos after exposed to vitrification solution at 1, 2, 3, 4 and 5 min were 84.0, 88.0, 71.0, 48.0 and 24.0% respectively.

• 한국양식학회지
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• 제20권4호
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• pp.260-264
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• 2007

본 연구에서는 대구 Gadus macrocephalus의 종묘생산을 위한 생물학적 기초자료를 제공하기 위하여 수온(4, 7, 10, 13 및 $16^{\circ}C$)에 따른 난발생, 자어의 성장 및 생존율을 조사하였다. 복부압박법으로 채란 하였으며, 채란양과 어체의 관계는 $F=80,220{\times}TL-3,397,700$ ($R^2=0.7528$)이었다. 수정란은 원형의 침성 점착란이었으며, 그 크기는 $0.90{\sim}1.11\;mm$였다. 습식법에 의한 수정률은 68%였다. 난발생의 각 단계에 이르기까지의 수온(T: $^{\circ}C$)에 따른 발생속도(t: hour)는 수온이 높을수록 빨랐으며, 그 관계식은 다음과 같았다. 4세포기: 1/t=0.0175T+0.0356 ($R^2=0.9579$), 16세포기: 1/t=0.0100T+0.0350 ($R^2=0.9759$), 상실기: 1/t=0.0083T-0.0181 ($R^2=0.9586$), 낭배기: 1/t=0.0035T-0.0058 ($R^2=0.871$), Kupffer씨 포 출현기: 1/t=0.0021T-0.0014 ($R^2=0.871$), 부화자어기: 1/t=0.0006T-0.0006 ($R^2=1$). 또한 대구의 난발생이 개시되는 생물학적 영도는 평균 $-0.4^{\circ}C$로 나타났다. 부화가능한 수온의 범위는 $7{\sim}10^{\circ}C$로 나타났다. 수정란에서 부화까지 소요되는 시간은 $7^{\circ}C$와 $10^{\circ}C$에서 각각 288시간(12일)과 192시간(8일)이었으며, 이때 부화율은 각각 65.0% 및 33.4%로 $7^{\circ}C$에서 3배정도 높았다. 부화자어를 40일간 $7^{\circ}C$와 $10^{\circ}C$에서 사육하였을 때, 성장에는 차이가 없었으나, 생존율은 각각 18.3% 및 5.2%로 $7^{\circ}C$에서 유의하게 높았다. 이상의 결론을 종합해 보면, 대구 종묘생산을 위한 수온은 $7^{\circ}C$가 적합하다고 판단된다.

• 생명과학회지
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• 제9권5호
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• pp.518-524
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• 1999

Timing of oviposition, nuclear maturation of oocytes, egg-sperm contact and brooding were observed in the giant fresh-water prawn Macrobrachium rosenbergii reared in the laboratory. When a pre-spawning molted female was transferred to the male, the female oviposited commonly at 5 to 10 hr after mating. The first polar body were extruded within 2-3 min after spawning. The egg chromosome progressed to release second polar body by 30 min and mitotic nuclear division occurred in 8 hr after spawning. Thumbtack-shaped spermatozoa were seen peneterating egg membrane with their spike directed forward. Transformation of spermatozoa and fertilization membrane could not be observed by light microscopy. The elapsed hours from oviposition to the end of brooding wee taken in 5-7 hr. Brood size for female 9 cm and 18 cm in body length was estimated to be 10,000~15,000 and 100,000 eggs, respectively and is proportional to the total length of the berried females. The ovigerous females reared at chlorinity of 2.21~4.25$\textperthousand$Cl. showed a normal egg development up to hatching.

• 한국수산과학회지
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• 제37권6호
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• pp.478-484
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• 2004

On 11 June 1991, eggs from the brood stock of small yellow croaker (Larimichthys polyactis) were artificially fertilized using the standard dry method and were hatched. Each of the fertilized eggs (1.1-1.2 mm in diameter) had an oil globule and was transparent and buoyant. The fertilized eggs hatched in a range of water temperatures $(17.5-20.3^{\circ}C)$ 44 hrs after fertilization. The total lengths of the newly hatched larvae were 3.1-3.3 mm, and these hatchlings had 31 myotomes (10+21). Melanophores and yellow-brown chromatophores were concentrated on the head, at the ventral part of the yolk, and in the middle of the tail. Four days after hatching, the larvae completely absorbed the yolk and became flexions of 5.1-5.5 mm in total length. Fifteen days after hatching, one spine (the anterior tip of the maxillary) appeared in the upper jaw and three spines developed at the upper parts of the eyes and on the posterior part of the head. At this stage, the larvae were approximately 8.3 mm long. Thirty-nine days after hatching, juveniles (1.9-3.4 mm in total length) had a pointed tail fin. By 66 days after hatching, the juvenile fish (about 4.0-6.5 mm in total length) were similar to adult fish in body shape. The larvae of L. polyactis could be distinguished from those of L. croacea by two distinct characteristics: the large number of vertebrae (28-29), and a relatively small bony ridge on the occipital region of the head.

• 한국발생생물학회지:발생과생식
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• 제20권1호
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• pp.31-40
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• 2016

This study was conducted in order to examine the egg development in red spotted grouper, Epinephelus akaara and the morphological development of its larvae and juveniles, and to obtain data for taxonomic research. This study was conducted in June 2013, and 50 male and female fish were used for the study. One hundred ${\mu}g/kg$ of LHRHa was injected into the body of the fish for inducing spawning, and the fish were kept in a small-sized fish holder ($2{\times}2{\times}2m$). Eggs were colorless transparent free pelagic eggs, 0.71-0.77 mm large (mean $0.74{\pm}0.02mm$, n=30), and had an oil globule. Hatching started within 27 h after fertilization. Pre-larvae that emerged just after hatching were 2.02-2.17 mm in total length (mean $2.10{\pm}0.11mm$), their mouth and anus were not opened yet, and the whole body was covered with a membrane fin. Post-larvae that emerged 15 days post hatching were 3.88-4.07 mm in total length (mean $3.98{\pm}0.13mm$), and had a ventral fin with two rays and a caudal fin with eight rays. Juveniles that were formed at 55 d post hatching, were 31.9-35.2 mm in total length (mean $33.6{\pm}2.33mm$), with red color deposited over the entire body, and black chromophores deposited in a spotted pattern. The number of fin rays, body color, and shape were the same as that in the adult fish.

• 한국발생생물학회지:발생과생식
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• 제18권1호
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• pp.13-23
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• 2014

This study is conducted to monitor the morphological developmental features of the egg development, larvae and juvenile of Epinephelus septemfasciatus, the fertilized eggs were gotton using artificial insemination. Matured parents are collected from marine caged fish farms in Geomun-ri, Samsan-myeon, Yeosu-si, Jeollanamdo Korea in June 2012. The fertilized eggs were pelagic eggs containing one oil globule, and measured 0.81~0.89 mm ($0.85{\pm}0.04mm$, n=50) in diameter. In regard to rearing environment, the water temperature is $21.0{\sim}23.0^{\circ}C$ and the salinity is 32.0~33.2‰. Hatching was observed from 48 hours after fertilization, the mouth and anus of prelarvae was not opened but had egg yolk at newly hatched. 4 days after hatching, the mouth and anus of postlarvae was opened and began to eat Rotifer and was measured 2.40~2.49 mm ($2.45{\pm}0.03mm$ n=10) in total length. 12 days after hatching, postlarvae was measured 3.77~4.67 mm ($4.27{\pm}0.33mm$) in total length, its the second pole tide of dorsal fin and the first pole tide of pelvic fin was extended longitudinally. 71 days after hatching, juvenile was measured 40.5~45.4 mm ($42.6{\pm}2.04mm$) in total length. Seven bands were observed in body, and pole tides of dorsal and pelvic fins were shortened.

• 한국환경생태학회지
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• 제22권1호
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• pp.59-65
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• 2008

본 연구는 2005년부터 2007년까지 파주, 강화도, 이천, 시화호, 충주 등 5개 지역에 서식하는 수리부엉이 9쌍의 둥지를 정기적으로 촬영, 관찰하여 번식생태와 교미행동을 연구하였다. 한배산란수는 $1\sim4$개로 평균 2.77개(n=15)였으며, 부화성공률은 약 71%(n=34)였고, 이소성공률은 약 83%(n=23)였다. 수리부엉이는 산란이 끝난 후에도 암수간의 교미행동이 지속되며, 새끼들이 둥지를 떠나는 시기인 부화 후 $7\sim8$주까지 교미행동을 계속한다는 것을 확인하였다. 이러한 수정 목적 이외의 교미 행동은 암컷과 수컷 모두의 필요에 의해서 이루어지는 것으로써 '부부관계(pair-bond)'의 확인, 유지, 강화를 통해 새끼를 안정적으로 키우기 위한 것으로 해석된다.