• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.41-41
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• 2001

Chromatin configuration and microtubule assembly were determined in porcine maturing and activated oocytes following intracytoplasmic sperm injection. Microtubule localization was confirmed using a mouse monoclonal antibody to $\alpha$-tubulin and detected using a fluorescent labeled goat anti-mouse secondary antibody. DNA was stained with propidium iodide. The image of microtubules and chromatin was captured using laser scanning confocal microscope. In germinal vesicle stage oocyte, sperm chromatin remained condensation and sperm derived microtubules were not observed at 8 to 12 h after sperm injection. At 24 h after injection, the sperm nucleus developed to the metaphase chromatin along the metaphase structure of female nucleus. In some metaphase I stage oocytes, sperm chromatin decondensed at 8 h to 12 h after injection, sperm aster was seen soon after sperm injection. At 24 h after sperm injection into metaphase I stage oocyte, male chromatin developed to the metaphase chromatin while female chromatin extruded first polar body and formed the metaphase chromatin. At 12 to 15 h after sperm injection into preactivated oocytes, condensed sperm nucleus was located in close proximity of female pronucleus. However, the condensed nucleus did not fuse with female pronucleus. In preactivated ocytes, injected sperm remained condensation, a few sperm organized small microtubular aster. Instead, maternal derived microtubules were organized near the female chromatin, which seem to move condensed male chromatin near to the female pronucleus. These results suggest that sperm nuclear decondensing activity and nucleation activity of centrosome during fertilization are cell cycle dependent. In absence of male functional centrosome, female origin centrosome takes over the role of microtubule nucleation for nuclear movement.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.18-18
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• 2001

Despite of importance of integrated events of nucleus and microtubule remodeling in nuclear transferred embryos with somatic cells, little information is available on this subject. In this study we configured chromatin and microtubule organization following somatic cell nuclear transfer in pre- and non-activated bovine oocytes in order to clearify nuclear remodeling process and to demonstrate centrosome inheritance during nuclear transfer. The cumulus-oocyte complexes were collected from slaughterhouse and were matured in vitro for 20 h in TCM 199 supplemented hormone. Matured bovine oocytes were enucleated by aspirating the frist polar body and metaphase chromatin using a beveled pipette. Bovine fibroblast cells were fused into enucleated oocyte by electrical stimulation. Reconstructed oocytes were activated with ionomycine and 6-dimethylaminopurin, and then cultured in CRlaa medium. The organization of nuclear and microtubules were observed using laser-scanning confocal microscopy. At 1 hour after fusion, microtubule aster was seen near the transferred nucleus in most oocytes regardless activation condition. While most of fibroblast nuclei remodeled to premature chromosome condensation (PCC) and to the two masses of chromosome in non-activated oocytes, a few number of fibloblasts went to PCC and multiple pronuclear like structures in activated oocytes. Microtubular spindle was seen around condensed chromosome. Gamma-tubulin was detected in the vicinity of condensed chromosome, suggesting this is a transient spindle. The spindle seperated nucleus into two masses of chromatin which developed to the pronuclear like structures. Two pronuclear like structures were than apposed by microtubular aster and formed one syngamy like nuclear structure at 15 h following nuclear transfer. At 17 to 18 h after fusion, two centrosomes were seen near the nucleus, which nucleates micrtubules for two cell cleavage. While 31% of reconstructed oocytes in non-activated condition developed to morulae and blastocysts, a few reconstructed oocytes in pre-activated condition developed to the blastocyst. These results suggested introduction of foreign centrosome during nuclear transfer, which appeared to give an important role for somatic cell nuclear reprogramming.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2001년도 추계학술대회 및 정기총회
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• pp.99-99
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• 2001

The acid-labile subunit (ALS) associates with insulin-like growth factor (IGF)-I or -II and IGF binding protein-3 (IGFBP-3) to form a 150-kD complex in the circulation. This complex is thought to regulate the serum IGFs by restricting them in the vascular system and promotes their endocrine actions. Little is known about how ALS binds to IGFBP3, which connects the IGFs to ALS. Xenopus oocyte was utilized to study the function of ALS in assembling IGFs into the ternary complexes. Xenopus oocyte was shown to correctly translate in vitro transcribed mRNAs of ALS and IGFBP3. IGFBP3 and ALS mRNAs were injected in mixture and their products were immunoprecipitated by antisera against ALS and IGFBP3. Contrary to the traditional reports that ALS interacts only with IGF-bound IGFBP3, this study shows that ALS is capable of forming a binary complex with IGFBP3 in the absence of IGF. When cross-linked by disuccinimidyl substrate, band representing ALS-IGFBP3 complex was evident on the PAGE. IGFBP3 movement was monitored according to the distribution between the hemispheres. Following a localized translation in the vegetal hemisphere, IGFBP3 was shown to remain in the vegetal half in the presence of ALS. Different from wild type IGFBP3, however, mutant IGFBP3 freely diffused into the animal half despite the presence of ALS. Taken together, this study suggests that ALS may play an important role in sequestering IGFBP3 polypeptides via the intermolecular aggregation. Studies using this heterologous model will lead to a better understanding of the IGFBP3 and ALS assembling into the ternary structure and circulating IGF system.

• 한국수산과학회지
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• 제33권3호
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• pp.219-224
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• 2000

난소의 내부는 결체성 조직인 다수의 난소박판으로 구성되며, 이곳에서 난원세포가 유래한다 정소조직상은 정세관 형태이며, 각각의 정세관은 여러 개의 소낭구조를 가진다. 각 소낭내의 생식 세포들은 같은 단계의 발달상태를 보인다. 군 성숙도에 도달하는 크기는 암${cdot}$수 모두 전장 4.5 cm이다. 암컷의 GSI는 6월에 가장 높은 값을 나타냈으며, 수컷의 GSI는 7월에 가장 높은 값을 나타냈다. 생식주기는 성장기 ($1{\~}3$월), 성숙기 ($4{\~}5$월), 완숙 및 산란기 ($6{\~}7$월) 그리고 회복 및 휴지기 ($8{\~}12$월)로 나눌 수 있었다. 난모세포 발달양식은 난군동기발달형에 속하며, 초기 성장기 난모세포의 세포질에서 난황핵이 관찰되었다.

• Applied Microscopy
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• 제20권2호
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• pp.117-126
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• 1990

Structural changes of the vitelline envelopes during oogenesis of a polychaete, Nectoneanthes oxypoda, were examined with a scanning electron micrscope. Oocytes grow in the same coelomic fluid to the final stage, but the surface appears to change in the structure during oogenesis. Projections, which were identified to be microvilli, change in shape, number and size. Short microvilli, which cover the surface of oocyte of $33{\mu}m$ diameter densely, grow in length, reaching a maximum at the stage of $73{\mu}m$. The number of microvilli increases with the stages of oogenesis, reaching a plateau at the stage of $82{\mu}m$. The observations suggest that control of material transport including yolk precursor proteins may be correlated with the structural changes in the microvilli.

• 한국동물학회지
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• 제38권2호
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• pp.212-219
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• 1995

Through the histolosical observation of ovary in the nine species of the genus Cobitis, we found that specific adhesive membranes were attached to the outer zone radials during the vitellosenesis. These adhesive membranes could be classified into four forms as follows: 1) granular form of Colitis lutheri, C. striate, and C. sinensis, 2) villous form of C. longicorpus, C. konensis koreensis, C. k. pumilus, and C. granoei, 3) saw-like form of C. rotundicaudata, and 4) filamentous form of C. chou. The various forms of adhesive membrane of egg in the cobitid fishes are showed a species specificitv with reference to their habitats and spawning property.

• 한국동물학회지
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• 제39권4호
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• pp.419-425
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• 1996

lksookimia(신칭,참종개속) 어류 5종의 부착막 구조 (adhesive membranes)를 조사하기 위하여 전자현미경으로 성숙란을 조사한 결과, 난황형성시기인 난황구 후기의 zona radiata에는 다음과 같은 3가지 형태의 부착막이 구별되었다. Iksookimia koreensis, I pumila, 그리고 I. hugowolfeldi는 융모형(villous form)을 보였으며, I.longicorpus는 원추모양의 솜 형태(cotton form)을 보여주었으며, I. choii는 덩클모양 (vine form)의 독특한 부착막 구조를 가지고 있었다. I. koreensis와 I pumila,그리고 I. hugowolfeldi는 같은 융모형의 부착막구조를 가지고 있으나 난막의 미세구조는 종간에 뚜렷한 차이를 보여주었다. 또한 참개종속 어류의 난막과 부착막 구조는 그들의 서식처와 산란습성에 관련된 종의 특이성을 보여 주고 있어서 분류학적으로 주목되었다.

• 한국발생생물학회지:발생과생식
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• 제25권3호
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• pp.123-131
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• 2021

We report on the gonadal structure abnormality and intersexuality found from a survey regarding the reproductive potential of the rock bream, Oplegnathus fasciatus, along the southern coast of Korea. In total, 448 samples were collected for histological analysis (total length 27.5±6.8 cm; total weight 522.9±385.6 g). The sex ratio (F:M) was 1:0.46 (n=307:141), with 68.5% being females. The frequency of oocyte atresia and ovarian neoplasm in the females was 85.3% and 21.5%, respectively, and the frequency of testicular lobule and cyst deformation and testicular neoplasm in the males was 73.1% and 37.6%, respectively. The scale formation in the ovary and testis was approximately 2.0% for both sexes. The intersexuality was 42.2%, with the intersexuality among females being 49.5%, which was significantly higher than that of males (26.2%). Such findings indicate a negative impact on the reproductive output of the rock bream along the southern coast of Korea; however, a more detailed study is required to ascertain accurate causes.

• 한국어류학회지
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• 제26권3호
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• pp.179-184
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• 2014

잉어과(Pisces, Cyprinidae)에 속하는 멸종위기 어류인 돌상어 Gobiobotia brevibarba의 난막미세구조 및 난자형성과정을 광학현미경과 주사전자현미경을 이용하여 관찰하였다. 2014년 5월의 돌상어 난소 내에는 다양한 발생단계의 생식세포들이 관찰되었다. 난모세포들의 상대면적은 성숙란(74.5%), 난황물질이 형성되는 난황구기(16.6%), 난황포기(6.6%), 그리고 미숙한 발달단계인 주변인기(2.3%) 순으로 구성되어 있었다. 주변인기 단계의 세포질은 hematoxylin에 강하게 염색되며, 핵막의 안쪽에는 다수의 인들이 존재하였다. 난황형성물질인 난황포기와 초기 난황구기 단계에서는 여포층과 방사대가 뚜렷하게 형성되기 시작되었으며, 방사대 위에는 미세융모의 난막구조가 더욱 뚜렷해졌다. 난황구기 및 성숙란의 단계에서는 난황구 일부가 융합되면서 강한 호산성을 보였다. 또한 성숙란의 표면에는 깔때기 모양의 난문이 관찰되었으며, $2{\sim}3{\mu}m$ 길이의 미세융모가 일정하게 분포하였다.

• 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.