• The Korean Journal of Zoology
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• v.24 no.1
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• pp.27-37
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• 1981

Protein patterns at various stages of pre-and post-fertilization in Drosophila melanogaster were analyzed using polyacrylamide gel electrophoresis. Ovaries at various times after hatching, embryos, larvae and pupae were collected and used for protein analysis. The protein patterns were found to vary qualitatively and quantitatively during oogenesis as well as during embryogenesis and metamorphosis. Yolk proteins, which were found to be synthesized in fat body and transferred into the oocytes, appeared in three bands in the gel in the early stages, but these proteins seemded to be converged into a single band in the late stage of oogenesis. The protein patterns in the post fertilization stages seemed to be abruptly changed at two different stages; between embryonic and larval stages and between the second and third instar stages. The pattern in the pupal stages was not very much different from that of the third instar.

• Korean Journal of Animal Reproduction
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• v.24 no.1
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• pp.59-67
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• 2000

This study was investigated the developmental potential of bovine embryos following nuclear transfer with bovine fetal fibroblasts (BFF). BFF were isolated from a male 45-day-old-fetus. Non-starved BFF labeled with MitoTracker were transferred into perivitelline space of enucleated oocytes. BFF-oocyte units were fused by electric pulse, and then fused oocytes were activated with calcium ionophore A23187 and subsequently 6-dimethylaminopurine (6-DMAP). The resulting zygotes were placed into CRlaa bovine embryo culture medium. Transfer of the nucleus into enucleated oocyte led to premature chromosome condensation, swelling and pronucleus formation. Remodeled oocytes were developed to the mitotic and 2-cell stage at 18 to 26 h after nuclear transfer. The incidence of in vitro development to the blastocyst stages was 21% of fused oocytes. Mitochondria of BFF eliminated rapidly and were not detected at 8 h after fusion. These results suggest that BFF can be successfully reprogrammed in enucleated bovine oocytes, and that reconstructed embryos can develop to the blastocyst stage.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.11 no.6
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• pp.2124-2128
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• 2010

In this study, we determined effects of the mitogen-activated protein kinase (MAPK) inhibitor, U0126 on meiotic maturation, microtubule organization and actin filament assembly in the porcine oocyte. The phosphorylated MAPK was first detected at 12 h after the initiation of maturation cultures, fully activated at 24h, and remained until metaphase II. Treatment of germinal vesicle (GV) stage oocytes with $20{\mu}M$ U0126 completely blocked MAPK phosphorylation, but germinal vesicle breakdown (GVBD) was normally proceeded. However, the oocytes didn‘t progress to the metaphase I. The inhibition of MAPK resulted in abnormal spindles. In oocytes treated with U0126 after GVBD, polar body extrusion was normal, but the organization of the metaphase plate and chromosome segregation were abnormal. In conclusion, MAPK activity plays an important regulatory role in GV chromatin configuration and meiotic progress in porcine oocyte maturation.

• Korean Journal of Veterinary Service
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• v.21 no.3
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• pp.267-275
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• 1998

돼지 수정란의 체외생산은 난자의 체외성숙과 체외수정에 관한 기술의 부족으로 아직까지 만족스럽지 못한 수준이다. 특히 돼지 수정란의 체외생산에는 복잡한 세포질의 성숙과정과 높은 다정자침입율 및 전핵형성의 억제등의 문제점이 있다. 본 연구에서는 돼지 난자의 체외성숙 체계를 개선하기 위하여 transforming growth factor$\beta$(TGF$\beta$)의 첨가가 난자 및 난구세포에 미치는 효과에 대하여 검토하였다. 체외성숙용 배지에 TGF$\beta$를 1~10ng/$m\ell$의 농도로 첨가하여 미성숙 난자를 배양한 결과 성숙율이 높아졌다. TGF$\beta$의 효과는 난구세포가 제거된 난자의 성숙에도 효과적이었다. TGF$\beta$(를 첨가하지 않은 배양액 내에서는 배양 24시간 까지 metaphase-II로 성숙된 난자가 관찰되지 않았으나 TGF$\beta$를 첨가한 배양액 내에서는 관찰되었다. 한편, 난구세포가 부착된 난자의 성숙배양시 TGF$\beta$의 첨가시기에 따른 차이는 없었으나, 난구세포를 제거한 난자의 경우에는 성숙배양 전반기(59%) 또는 후반기(57%) 24시간 동안에만 TGF$\beta$를 첨가하는 것이 48시간 동안 계속하여 첨가(27%)하는 경우 또는 비첨가(38%)에 비하여 유의적으로 높은 성숙율을 나타냈다(p<0.05). 이와 같은 결과는 난구 세포가 돼지 난자의 체외성숙에 필수적이지만 TGF$\beta$는 난구세포가 제거된 난자의 체외성숙에 어 느정도 유익한 효과를 발휘하는 것으로 추측된다.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 1998.07a
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• pp.63-65
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• 1998

생쥐의 초기발생과정에서 배아 세포질의 제거로 nucleus/cytoplasm ratio를 증가시켰을 경우, 발생과 gene expression에는 영향을 미치지 않았으나 형태형성은 촉진되었다. 또, nucleus/cytoplasm ratio의 변화 없이 발생후기의 cytoplasm으로 일부 치환된 배아에서 발생과 gene expression, 형태형성은 모두 촉진되었다. 이상의 결과로 보아 초기배아의 gene expression과 형태형성과정에서 일어나는 time schedule의 조절은 nucleus/cytoplasm ratio가 직접적으로 영향을 미치지 않고 난자와 배아 내에 존재하는 cytoplasm factor에 의해 조절 받으며, 특히 형태 형성의 time schedule은 cytoplasmic factor와 이들에 의해 조절되는 물질의 상대적인 농도에 의해 조절될 것으로 여겨진다.

• The Korean Journal of Zoology
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• v.31 no.4
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• pp.318-326
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• 1988

여포세포에서 합성된 난황단백질이 난모세포로 이동하는 동안에 난황체가 이 두 종류의 세포사이에 형성되었다가 결국은 난황막으로 전환한다. 단계7까지는 뚜렷하게 보이던 난모세포막과 여포세포막이 소멸되고 그 자리에 전자 밀도가 높은 난황체 물질이 산만하게 축적된다. 난황체는 단계9에서 막성 구조의 일종인 linkage bridge로 둘러싸여 단계11까지는 두께가 5∼7um가 되리 만큼 성숙한다. 단계13에서 난황체는 비로소 난황막으로 전환되는데, 이때 난황막의 두께는 겨우 1 U m에 지나지 않는다. 이러한 두깨의 감소는 난황체 물질이 다량 난모세포 쪽으로 이동한 것으로 생각 되었다. coated vesicle을 포함한 다양한 종류의 과립이 난황체 양쪽에서 관찰되었는데, 난모세포쪽에 출현한 과립은 난황체 물질이 난모세포로 이동되는 구조로 해석되었고 여포 세포쪽에서 관찰된 과립은 주로 난황체의 전자밀도와 동일한 점으로보아 여포세포에서 합성되어 난황체를 형성하는 물질로 이루어진 구조로 해석되었다. As yolk proteins are transported from !he follicle cells into oocvtes, vitelline body forms and changes into a vitelline membrane between the ko celt types during the vitellogenic period. Cell membranes of oocyte and follicle cells surrounding the oocyte disappear at stage 7 and high electron-dense substance of vitelline body simultaneously accumulates sporadically between the cell types. The vitelline body becomes surrounded by linkage bridge, a membranous structure, at stage 9 and greatly increases in thickness to be 5-7 U m thick at stage 11. At stage 13 the vitelline body becomes vitelline membrane, which is now only 1 U m thick, suggesting that much of the substance of the vitelline body has been transported into oocyte. Various types of vesicles including coated vesicles were observed at both sides of th vitelline body. The vesicles occurred at the side of oocyte were interpreted to be structures transported from the vitelline body into oocyte, whereas those found at the side of the follicle cells were thought to be structures made in the follicle cells and fused into the vitelline body.

• Applied Microscopy
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• v.41 no.3
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• pp.163-168
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• 2011

Tiger barb (Puntius tetrazona Bleeker, 1855) is a teleost belonging to Cyprinidae. The oogenesis of tiger barb was investigated by light microscope. The ovary was of white and ellipsoidal shape with the major axis 2 cm and the minor axis 1 cm. Cytoplasm of oogonia was basophilic and many nucleoli were located at inside of nuclear membrane. In early stage of primary oocyte, yolk vesicles were distributed only in the marginal area. In secondary oocyte, the egg envelope was formed and yolk vesicles was formed in the cytoplasm. The basophilic substance of cytoplasm was changed to acidic. In case of matured egg, thickness of egg envelope and size of egg were increased. The yolk vesicles were changed to yolk mass in accordance with development. Also, there is not the formation of oil droplets in cytoplasm. In conclusion, the oogenesis of tiger barb was characterized by the increase in cell size, the formation of yolk, the decrease of basophilic substance in the cytoplasm, and formation of yolk mass.

• Korean Journal of Animal Reproduction
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• v.24 no.2
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• pp.143-154
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• 2000

Most eggs initiated the fertilization processes but arrested at specific stages. The stages included failure of the oocyte to exit from the meiotic metaphase-II with or without sperm penetration, failure of appropriate sperm aster formation, inability to form proper male and female pronuclei, failure of suitable pronuclear apposition, and failure to form proper number of either male or female pronuclei. Various images of defective microtubule organization and chromatin configuration during IVF and ICSI procedures were observed. We discussed the data with previous research results during normal fertilization in humans and other mammals. In conclusion, various aberrant patterns in microtubule assembly and chromatin configuration, which were assessed in the present study, could be used as criteria to improve assisted reproductive technology in clinics. However, further cellular and molecular characterization is needed to clarify these aberrant patterns of cytoskeletal assembly.

• Applied Microscopy
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• v.40 no.2
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• pp.65-71
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• 2010

Kribensis, Pelvicachromis pulcher is a teleost belonging to Cichlidae. The oogenesis was investigated by light microscope. The ovary was located between intestine and air bladder, a yellowish and ellipsoidal shape with the major axis 20mm and the minor axis 5 mm. Cytoplasm of oogonia in early stage was basophilic and many nucleoli were located at inside of nuclear membrane. In primary oocytes, yolk vesicles were distributed only in the marginal area and egg envelope was not formed on the outside of an egg. In secondary oocytes, the egg envelope was formed and yolk vesicles in the cytoplasm were increased than the earlier stage. The basophilic substance of cytoplasm was changed to acidic. Some yolk vesicles started forming small yolk mass except the surrounding nucleus. In case of matured egg, size of egg were increased. The yolk vesicles were changed to yolk mass in accordance with development. The yolk mass contained crystal-like structures. In conclusion, the oogenesis of Pelvicachromis pulcher was summarized by the increase in cell size, the formation and the accumulation of yolk, and the decrease of basophilic substance in the cytoplasm. The oogenesis of Coreoleuciscus splendidus is similar with other teleost. But there were differences in distribution of yolk vesicle and yolk mass containing cristal-like structures.

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.201-207
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• 2017

To increase the productivity of in vitro development, the antioxidants have been used for culture system of bovine oocytes and embryos. However, comparative studies on these molecules are rare and direct beneficial effects on blastocyst production cannot be discriminated for best results. The study was conducted to determine the influence of N-acetyl-L-cysteine (NAC), N-acetyl-L-cysteine amide (NACA), glutathione (GSH) and cysteamime (CYS) on maturation competence of COCs from GV to MII stage and productivity of blastocyst formation during in vitro fertilization and culture. There was no difference among maturation rates of oocytes to metaphase II with polar body with antioxidants for any of the treatment groups (p>0.05). However, the significant improvement on the rate of blastocysts ($32.3{\pm}5.0%$) was found in 0.1 mM CYS treatment than 0.3 mM NAC, 0.2 mM NACA or 0.5mM GSH (p<0.05). The addition of NAC ($18.8{\pm}3.7%$) or NACA ($21.2{\pm}3.9%$) did not improve development competence to morula and blastocysts than control ($24.4{\pm}4.1%$) and GSH ($26.5{\pm}5.0%$) (p>0.05). Our study showed that medium supplementation with CYS during IVM and IVC improved the rate of bovine embryo development but not with NAC, NACA and GSH addition.