• 제목/요약/키워드: Polar bodies

검색결과 38건 처리시간 0.022초

Production and development of porcine tetraploid parthenogenetic embryos

  • Lin, Tao;Lee, Jae Eun;Shin, Hyeon Yeong;Lee, Joo Bin;Kim, So Yeon;Jin, Dong Il
    • Journal of Animal Science and Technology
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    • 제61권4호
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    • pp.225-233
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    • 2019
  • The aim of this study was to produce porcine tetraploid (4N) parthenogenetic embryos using various methods and evaluate their developmental potential. In method 1 (M1), porcine 4N parthenogenetic embryos were obtained by inhibiting extrusion of both first (PB1) and second (PB2) polar bodies; in methods 2 (M2) and 3 (M3), 4N parthenogenetic embryos were obtained by electrofusion of 2-cell stage diploid parthenogenetic embryos derived from inhibition of PB2 or PB1 extrusion, respectively. We found no differences in the rates of cleavage or blastocyst formation or the proportion of 4N embryos among M1, M2, and M3 groups. The different methods also did not influence apoptosis rates (number of TUNEL-positive cells/number of total cells) or expression levels of apoptosis-related BAX and BCL2L1 genes. However, total cell and EdU (5-ethynyl-2'-deoxyuridine)-positive cell numbers in 4N parthenogenetic blastocysts derived from M1 were higher (p < 0.05) than those for M2 and M3 groups. Our results suggest that, although porcine 4N parthenogenetic embryos could be produced by a variety of methods, inhibition of PB1 and PB2 extrusion (M1) is superior to electrofusion of 2-cell stage diploid parthenogenetic embryos derived from inhibition of PB2 (M2) or PB1 (M3) extrusion.

High efficiency of homemade culture medium supplemented with GDF9-β in human oocytes for rescue in vitro maturation

  • Mohsenzadeh, Mehdi;Khalili, Mohammad Ali;Anbari, Fatemeh;Vatanparast, Mahboubeh
    • Clinical and Experimental Reproductive Medicine
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    • 제49권2호
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    • pp.149-158
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    • 2022
  • Objective: Optimizing culture media for the incubation of immature oocytes is a vital strategy to increase the oocyte maturation rate during in vitro maturation (IVM) programs. This study evaluated the IVM and fertilization rates of human germinal vesicle (GV) and metaphase I (MI) oocytes using two different maturation media (commercial and homemade) with or without growth differentiation factor 9-β (GDF9-β). supplementation. Methods: Immature oocytes from intracytoplasmic sperm injection (ICSI) cycles were collected and assigned to one of two IVM culture media (commercial or homemade; cleavage-stage base). After maturation, MII oocytes were examined under an inverted microscope for the presence of the polar body, zona pellucida (ZP) birefringence, and meiotic spindle (MS) visualization after maturation in four conditions (commercial or homemade medium, with or without GDF9-β. ICSI was done for matured oocytes, and fertilization was confirmed by the visualization of two distinct pronuclei and two polar bodies. Results: No significant differences were found between the two culture media in terms of the time and rate of oocyte maturation or the rate of fertilization (p>0.05). Growth factor supplementation increased the 24-hour maturation rate for both GV and MI oocytes only in homemade medium. The maturation rate after 24 hours was higher for MI oocytes (p<0.05). Similar results were observed for MS visualization and ZP structure in both types of media (p>0.05). Conclusion: Higher rates of oocyte maturation and fertilization were observed after application of homemade medium supplemented with GDF9-β. Therefore, this combination may be recommended as an alternative for clinical IVM programs.

In Vitro Development and Apoptosis in Haploid, Diploid Parthenotes and Fertilized Embryos

  • Y. J Chung;Lee, H. Y.;S. H. Jun;X. S. Cui;Kim, N. H.
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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    • pp.28-28
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    • 2003
  • Haploid parthenotes have been shown to be developmentally delayed compared with diploid parthenogenetic embryos in the mouse and pig. These developmental defects have been hypothesized to rusult from insufficient parthenogenetic activation, suboptimal in vitro culture conditions, or genemic imprinting. In the present study we compared the incidence of apoptosis and apoptosis related gene expression in pig haploid, diploid parthenotes and fertilized embryos. In vitro matured porcine oocytes were activated by electrical stimulation. Haploid activated oocytes with two polar bodies under stereomicroscopy were defined haploid parthenotes, oocytes with one polar body were defined as diploid parthenotes after 3h cycloheximide teatment. The morphological analysis of apoptosis in embryos was carried out using propidium iodide staining and terminal deoxynucleotidyl transferase mediated dUTP nick end labeling. The expression of Bcl-xL, Bak and P53 in haploid, diploid and in vivo fertilized blastocysts was determined using RT-PCR. Lower number of the haploid pig parthenotes developed to the morulae and blastocysts compared to the diploid parthnotes. Number of cells significantly lower in the haploid-derived blastocysts than diploid-derived it. Developmentally retarded haploid parthenotes exibited apoptosis at a significantly higher frequency than did diploid parthenotes and fertilized embryos. Level of Bcl-xL expression, diploid parthenotes similar to in vivo-derived it was higher than haploid parthenotes. However, Bak and P53 mRNA expression were not different among haploid, diploid, and fertilized embryos. This result suggested that parthenogenetic activation and parthenogenesis themselves do not cause apoptosis, but haploid increases the incidence of apoptosis in preimplantation embryos. Apoptosis may be due to decrease expression of Bcl-xL in haploid parthenotes developing in vitro.

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부이 특성에 따른 궤도 차량 동적 거동 (Dynamic Analysis of Tracked Vehicle by Buoy Characteristics)

  • 김형우;민천홍;이창호;홍섭;배대성;오재원
    • Ocean and Polar Research
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    • 제36권4호
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    • pp.495-503
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    • 2014
  • This paper focuses on the dynamic responses of a tracked vehicle crawling on extremely cohesive soft soil, each side of which is composed of two parallel tracks. The tracked vehicle consisted of 2 bodies. One body is the tracked vehicle body, which is assumed to be a rigid body with 6 DOFs. The other body is the buoy body. The two bodies are connected by a revolute joint. In order to evaluate the travelling performance of a 7 DOFs vehicle, a dynamic analysis program for the tracked vehicle was developed using Newmark's method and the incremental-iterative method. The effects of road wheels on the track and soil are not taken into account. A terra-mechanics model of extremely cohesive soft soil is implemented in form of relationships: normal pressure to sinkage, shear resistance to shear displacement, and dynamic sinkage to shear displacement. Pressure-sinkage relationship and shear displacement-stress relationship should represent the non-linear characteristics of extremely soft soil. Especially, since the shear resistance of soft soil is very sensitive to shear displacement, spatial distribution of shear displacement occurring at the contact area of the tracks should be calculated precisely. The proposed program is developed in FORTRAN.

칠레 남부 라고 소피아 (Lago Sofla) 심해저 하도 역암의 층구조와 퇴적 스타일 (Architecture and Depositional Style of Gravelly, Deep-Sea Channels: Lago Sofia Conglomerate, Southeyn Chile)

  • 최문영;조형래;손영관;김예동
    • 한국석유지질학회지
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    • 제10권1_2호
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    • pp.23-33
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    • 2004
  • 칠레 남부에 분포하는 라고 소피아 역암 (후기 백악기)은 이암이 우세한 심해 퇴적층 (Cerro Toro Formation) 내에 렌즈상으로 산출하고 남북방향으로 120 km 이상 연장된다. 라고 소피아 역암은 융기대전면분지 (foreiand basin)인 마젤란 분지의 융기대전면곡분 (foredeep trough)을 따라 발달한 심해저 하도계 퇴적층으로 해석된다. 이처럼 연장이 대단히 좋은 역질의 심해저 하도가 발달하는 것은 현생 심해저 환경에서 매우 드문 현상으로 라고 소피아 역암은 퇴적학적으로 매우 흥미로운 예이다. 연구지역의 북부에 분포하는 라고 소피아 역암은 이암 퇴적층 사이에 협재하는 3-5매의 역암체로 구성되고, 고수류 측정에 따르면 퇴적물은 동, 남, 남동 방향으로 운반된 것으로 유추된다. 이 역암체는 융기대전면곡분의 서편에 위치한 해저사면을 따라 발달한 심해저 하도계의 지류에서 퇴적된 것으로 해석되며, 지류들은 남북방향의 주하도로 수렴하였을 것으로 추정된다. 남부 지역의 라고 소피아 역암은 300 m 이상의 두께를 가지는 역암체로 구성되고, 남북방향으로 긴 융기대전면분지의 축을 따라 발달한 주하도에서 퇴적된 것으로 해석된다. 이 역암체는 층리를 보이는 역암, 괴상 혹은 점이층리의 역암, 기질지지 역암으로 구성되며, 각각은 저탁류에 의한 밑짐 운반, 고밀도 저탁류, 니질 암설류에 의해 퇴적된 것으로 해석된다. 층리역암에서 측정된 고수류 방향은 남남서항으로 주하도의 방향을 지시한다. 반면, 북부 및 남부 지역의 기질지지 역암에서 측정된 고수류 방향은 흔히 하도 방향에 대해 고각도를 이루는데, 이는 하도의 둑 또는 주변 사면이 붕괴하여 니질 암설류가 형성되었음을 지시한다. 형태구성 (architecture) 분석 결과, 라고 소피아 역암은 육상의 역질 망상하천 퇴적층과 유사한 구성요소로 구성되며, 라고 소피아 심해저 하도계는 망상하천과 유사한 지형적 특성을 지녔을 것으로 추정된다. 또한 하도 역암 내 큰 규모의 층구조는 동쪽으로 이동 누적된 특징을 보이는데, 이는 지구조 운동에 의해 주하도가 점진적으로 동쪽으로 이동하였을 가능성을 시사한다.

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U. unicinctus 난자의 인공수정과 감수분열 장치의 회전-이동행위에 관한 면역형광현미경 및 전자현미경적 연구 (Immunofluorescence and Electron Microscopic Study on the Artificial Insemination and Rotation-Shift Behaviors of the Bipolar Spindle Fiber in U. unicinctus Egg)

  • 권혁제;정진욱;김완종;신길상
    • Applied Microscopy
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    • 제33권2호
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    • pp.105-116
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    • 2003
  • U. unicinctus 난세포를 인공수정한 결과 수정 시기는 germinal vesicle (2n)기였으며 정자 중앙부에서 돌출한 actomere와 난모세포의 미세융모 끝에서 세포막 융합이 시작되는 것으로 관찰되었다. Germinal vesicle기에 수정이 가능하였으므로 pre-mitotic spindle이 관찰되지 않은 것으로 사료되었다. 수정 후 난모세포는 제1, 2감수분열을 수행하였으며 각각의 감수분열 장치들은 감수분열 말기에 난모세포막과 밀접한 구조를 형성하였고, 이 부위에서 극체가 형성되는 것을 볼 수 있었다. 극체 형성시 난모세포막은 세포분열의 관점에서 형태가 없는 것이 아니라 항-튜블린-FITC에 대한 활성구조를 형성하였으며, 각 감수분열 장치의 한쪽 극 (pole)과 어떤 복합구조를 형성하는 것으로 보인다. 제2극체 형성도 제1극체와 유사한 방법으로 형성되었으나, 제2감수분열 장치는 난모세포막의 접선에 평행하게 생성된 후 세포막을 향해 이동하면서 방추사의 양극성이 회전 (shift-rotation)하였고 접선에 수직으로 정렬하는 것을 볼 수 있었다. 난모세포의 감수분열 장치에서 방추사의 활성은 강하였으나 aster의 활성은 비교적 약한 것으로 보였다. 제2감수분열이 진행되는 동안 난모세포질에 머믈고 있던 정자는 점차 미래의 자성 전핵 형성부인 난모세포막 근접부로 이동하는 것이 관찰되었다. 난모세포막 근접부로 이동하는 동안 정자 aster의 활성은 점차 강해지는 반면 난모세포의 aster에서는 활성이 미약한 것으로 보아서 자웅 전핵융합을 주도하는 것은 정자 유래의 aster인 것으로 사료되었다. 제1난모세포의 감수분열 장치가 활성화되는 시기에 제1극체의 방추사에서도 강한 활성을 관찰할 수 있었다.

Effect of Supplementation of Vitamin E on In Vitro Maturation and Activation of Bovine Oocytes

  • Park, Jong-Im;Jang, Yun-I
    • 한국수정란이식학회지
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    • 제25권3호
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    • pp.165-169
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    • 2010
  • This study was carried out to assess the effect of vitamin E against the reactive oxygen species (ROS) on chemical activation of in vitro matured oocytes. Bovine oocytes were aspirated from slaughtered ovaries and transferred to maturation medium with or without vitamin E ($100\;{\mu}M$). After 22 hours of culture, oocytes with polar bodies were selected and submitted to activation treatments with or without vitamin E. After activation, oocytes were cultured in mSOF medium and rate of development was monitored. For ROS ($H_2O_2$) detection, in vitro matured and activated oocytes were selected and stained with DCFDA and observed under fluorescence microscope. The ROS contents were not significant differences in IVM rate, activation process and embryonic development to blastocysts with or without vitamin E. The cell number of blastocyst showed significant difference (p<0.05) in embryos matured and activated with vitamin E. The results of the present study demonstrated that the exposure of vitamin E in IVM and activation process improved the quality of embryos evaluated by the cell number of blastocysts.

한국 연안역 수질성분들(COD, DIN, DIP, Chlorophyll-a)의 해역별 농도 특성과 등급화: 통계적 접근 (Concentration Level and Grading of Water Quality Components (COD, DIN, DIP, Chlorophyll-a) in Korean Coastal Waters: A Statistical Approach)

  • 임동일;최현우;김영옥;정회수;강영실
    • Ocean and Polar Research
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    • 제33권1호
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    • pp.13-20
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    • 2011
  • Comprehensive data collection was carried out between 2001-2006 to investigate the concentration levels of chlorophyll-a, nutrients (DIN and DIP), and COD for surface waters of Korean coastal areas. A statistical analysis of these parameters was carried out on the basis of the frequency distribution of their concentration. Furthermore, the numeric grading for chlorophyll-a, DIN, DIP, and COD concentrations were derived statistically from the normalized frequency distribution of log-transformed data. The statistical grading clearly reflects the water quality characteristics of three Korean coastal water bodies (Western, Southern and Eastern coastal zones), which indicate common environmental and ecological characteristics. So, this study could provide useful information to set up the guideline for water quality assessment of Korean coasts.

토끼 핵이식에 의한 복제산자의 생산효율 향상에 관한 연구 (Production of Identical Rabbit Offspring by Nuclear Transplantation)

  • 박충생;윤희준;조성근;노규진;이효종;최상용
    • 한국수정란이식학회지
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    • 제14권3호
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    • pp.195-201
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    • 1999
  • This study was carried out to improve a technique of cloned animal prodcution by preactivation of nuclear recipient oocytes with ionomycin and 6-dimethylaminopurine (6-DMAP) in rabbits. The oocytes were collected from the oviduct of superovulated rabbit at 19∼20 hours post hCG injection. The collected oocytes were preactivated and self-enucleated by treating 5 uM ionoycin for 5 min, and 2.0 mM 6-DMAP for two hours. Microsurgical removel of the chromation complex in the second polar bodies was effectively performd and single blastomere separated from 32-cell stage rabbit embryos was injected into the perivitelline space of the enculeated recipient oocyts. Follwoing electrofusion and in vitro culture for 18 hours, the nuclear transplant(NT) embryos were transferred into the uterine horns of naturally mated or synchronized recipient does. When 32 NT embryous reconstituted with preactivated oocytes were transferred to 2 recipient does, one foster doe delivered two offspring (6.3%), while not a offspring was delivered from three foster does which received 17 NT embryos reconstituted with non-preactivated oocytes. A total of 68 NT embryos reconstituted with preactivated oocytes were transferred into the uterine horns of 7 synchronized ecipient does. Among them, two recipients were pregnant and delivered three offspring(5.9%).

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저회의 계면 화학적 특성 규명 (Characterization of interfacial chemistry on the coal bottom ash)

  • 이기강
    • 한국결정성장학회지
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    • 제21권2호
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    • pp.92-97
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    • 2011
  • 화력발전소에서 발생하는 석탄회의 일종인 저회는 현재 적절한 재활용 방법이 연구 되지 않아 대부분 매립에 의존하고 있는 실정이다. 본 연구에서는 비가소성 분체인 저회의 재활용율을 높이며, 저회를 점토 대체 원료로 재활용하기 위하여 저회와 점토의 계면화학적 성질을 조절하여 Bottom-ash(B/A)-Hard Clay(H/C) 소지의 물성변화를 관찰하였다. H/C 100 %, H/C60 % + B/A40 %에 대하여 각 조성별 pH에 따른 침강높이를 관찰한 결과 점토와 저회는 계면 화학적 성질이 비슷하여 혼합하여 슬립제조 시 hetero-polar 응집현상이 없었다. 또한 각 조성별 점도가 조절된 Slip을 주입 성형하여 $1100{\sim}1250^{\circ}C$에서 $50^{\circ}C$ 간격으로 소결하여 각 시편에 대한 비중, 흡수율을 측정하고 미세구조를 관찰하였다. 그 결과 분산된 점토와 저회 슬립은 Bingham 가소성 거동을 보이므로 주입성형 및 가소성형이 가능하며, 분산된 점토와 저회 슬립으로부터제조된 소결체는 $1250^{\circ}C$에서 KSL 4201과 KSL 1001의 규격을 만족할 뿐 아니라 특히 비중이 약 15 % 이상 경량이므로 도자기나 위생도기 등의 소지로 응용 가능성이 있다고 사료된다.