• The Korean Journal of Zoology
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• v.35 no.1
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• pp.37-44
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• 1992

북방산개구리, 참개구리 및 옴개구리를 사용하여 성숙된 난자의 세포질에서 활성을 띠는 성숙촉진요인( maturation promoting factor, MPF)을 미세주입 법으로 확인하고 이들의 성질을 조사하였다. 핵붕괴(germinal vesicle breakdown, GVBD)된 난자의 세포질을 미성숙 난자(GV난자)에 주입하고(75-100 nl) 이들을 15-24시간 배양했을 때 대부분의 GV 난자들이 핵붕괴를 일으켰으나(약 80%) 미성숙 난자(GV 난자)의 세포질을 주입했을 때에는 약 200nl의 난자들만이 핵붕괴를 일으켰다. 핵붕괴된 난자들의 crude extract를 주입했을 때에도 역시 성숙유도 효과가 있었으며 이종간에도 효과가 있었다. 난자의 성숙을 잘 일으키지 않는 옴개구리의 난자를 사용하여 MPF를 가진 세포질을 계대주입(serial transfer)하였을 때에도 MPF가 계속 활성을 띠는 것을 확인할 수 있었다. 아울러 개구리 난자의 MPF생성과 증폭과정에 CAMP의 증가나 단백질 합성의 저해가 미치는 영향을 조사한 결과. MPF의 작용이 유의하게 이들에 의해 억제되는 것을 알 수 있었다.

• Development and Reproduction
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• v.4 no.1
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• pp.7-12
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• 2000

At fertilization, sperm penetrates into oocyte, male and female pronuclei are fused together, and mitotic division follows. However, little information is available on the interactive roles and dynamic processes between cytoplasmic and nuclear components during the pronuclear formation, migration and cell division. The assisted reproductive technologies such as, intracytoplasmic sperm injection (ICSI) and round spermatid injection(ROSI) could provides new treatments for the male infertility as well as tools for the study of basic mechanism during fertilization. Nuclear transfer can also provide a mechanism on the interactive roles between nucleus and cytoplasm since the process includes nuclear reprogrammming of differentiated cells in the enucleated oocytes. Recently, I have investigated developmental processes in porcine oocytes following fertilization parthenogenesis, ICSI, ROSI and nuclear transfer using indirect immunocytochemical and electron microscopic studies. The results could provide an insight into biological questions related with epigenesis as well as strategies for the enhancement of embryology in general such as ICSI and nuclear transfer.

• Proceedings of the KSAR Conference
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• 2002.06a
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• pp.38-38
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• 2002

돼지 체세포 핵이식에 있어서 체외발달률을 재고하기 위하여 공여세포의 종류, 활성화 방법, 배양조건 등이 체외발달에 미치는 영향을 검토하였다. 도축장에서 채취한 난소에서 미성숙난자를 채취하여 NCSU-37 배양액에 10% pFF 와 0.6mM Cystein, 1mM dbcAMP, 0.1 IU/㎖ human menopausal gonadotrophin(hMG)에서 20시간 배양후 dbcAMP 와 hMG 가 없는 배양액에서 18-24 시간 추가배양 성숙 후 성숙된 난자를 핵이식의 수핵난자로 사용하였다. (중략)

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.2
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• pp.148-152
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• 2000

To provide basic information on the integumentary system of the blenny, Pholis nebulosa, ultrastructures of epidermal and dermal layer were examined by means of the light and transmission electron microscope. The skin of the blenny consisted of epidermal and dermal layer. Epidermal layer consisted of supporting cell and unicellular gland. The supporting cells were classified into superficial cell, intermediated cell and basal cell, and the gland cells were classified into mucous secretory cell and club cell. The cytoplasm of supporting cells was divided into cortex and medullar part. In the cortex and medullar part, microfilaments and cell organelles were well developed, respectively. Superficial cell of epidermal layer was cuboidal and contained nucleus of horseshoe shape. Intermediated cell had a nucleus of irregular form and the electron density was higher than the other supporting cells, Basal cell was columnar, but nucleus was situated in the upper cytoplasm. Cell organelles of the basal cell were poor than the other supporting cells, but membrane interdigitations were well developed. The cytoplasm of mucous secretory cell had a well-developed ovoid secretory granules, which reacted to red with AB-PAS reaction. The club cell had a we31-developed round secretory granules and endoplasmic reticulum. figment cells were classified into two type. The one contained pigment granules of electron dense, and the other contained reflecting platelets. The cytoplasm of fibrocyte had n well developed rough endoplasmic reticulum.

• Reproductive and Developmental Biology
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• v.28 no.1
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• pp.1-6
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• 2004

This study was conducted to investigate the effects of donor cell type, individual, passage number and trypsinization time on the in vitro development of bovine somatic cell nuclear transfer embryos. Three cell types (skin, muscle and cumulus cells) and cells from 3 individuals were used for nuclear transfer. Cell were passaged by 5, 15 or 30 times, and cell were trypsinized for 1 or 3 min before injection. Nuclear transfer were performed by conventional fusion method. Development rates to the blastocyst stage were not significantly different among three cell types (16.5∼23.9%) and individuals (16.4∼19.5%). Blastocyst formation rate of cloned embryos reconstituted with cells at passage 30 (5.8%) was significantly lower than those of embryos reconstituted with 5- and 15-passaged cells (25.3 and 23.5%, respectively, P＜0.05). The rate of embryos developed to the blastocyst stage was higher in embryos reconstituted with cells trypsinized for 1 min (30.7%) compared to embryos reconstituted with cells trypsinized for 3 min (P＜0.05). The result of the present study indicates that different donor cell types and individuals used in this study did not affect the development of cloned bovine embryos. However, passage number and trypsinization time of donor cells affect the in vitro development of cloned bovine embryos.

• Journal of Korea Multimedia Society
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• v.10 no.7
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• pp.846-855
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• 2007

Significant feature extraction in cancer cell image analysis is an important process for grading cell carcinoma. In this study, we propose a method for 3D quantitative analysis of cell nuclei based upon digital image cytometry. First, we acquired volumetric renal cell carcinoma data for each grade using confocal laser scanning microscopy and segmented cell nuclei employing color features based upon a supervised teaming scheme. For 3D visualization, we used a contour-based method for surface rendering and a 3D texture mapping method for volume rendering. We then defined and extracted the 3D morphological features of cell nuclei. To evaluate what quantitative features of 3D analysis could contribute to diagnostic information, we analyzed the statistical significance of the extracted 3D features in each grade using an analysis of variance (ANOVA). Finally, we compared the 2D with the 3D features of cell nuclei and analyzed the correlations between them. We found statistically significant correlations between nuclear grade and 3D morphological features. The proposed method has potential for use as fundamental research in developing a new nuclear grading system for accurate diagnosis and prediction of prognosis.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.65-65
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• 2001

본 연구는 칡소 귀세포를 공여핵으로 이용한 체세포 복제송아지 생산에 있어서 배양방법이 배발생 및 배반포 발달율에 미치는 영향과 체세포 복제란의 이식후 수태율에 미치는 영향을조사하여 복제송아지의 생산 효율을 제고하고자 실시하였다. 실험에 공시된 공여핵은 칡소 의 귀세포를 회수하여 10%FBS가 첨가된 DMEM배지에서 3-4일 동안 배양하여 monolayar Confluent 형성 후 0.25% trypsin을 처리하여 준비하였으며 공여세포는 적어도 passage가 5회 이상의 세포만을 사용하였다. 복제수정란의 생산은 18-20시간 동안 체외성숙 된 난자의 핵을 제거하고 공여핵을 주입하여 2.2kv/cm, 10$\mu\textrm{s}$의 전압으로 2회 자극함으로 융합하였으며, 융합된 난자는 5$\mu\textrm{g}$/$m\ell$ ionomycin에서 4분간, 1.9mM 6-dimethyl aminopurine에서 4시간동안 배양하여 활성화처리를 하였다. 핵이식수정란의 배양은 39$^{\circ}C$, 5%$CO_2$ incubator에서 처리구 I은 CRlaa에서 4일간 배양 후 CR2aa배지에서 배양, 처리구II는 CRlaa에 4일간 배양후 CR2aa배지에 cumulus cell과 공배양, 처리구III은 CR2aa 배지에 camulus cell과 함께 배양하였다. 수정란이식은 발정발현 7일째에 비외과적 방법으로 젖소 미경산우에 이식하였으며 이식란수는 2~4개의 핵이식된 수정란을 이식하였다. 임신진단은 45~60일 사이에 직장검사 및 초음파 진단기를 이용하여 실시하였다. 배양방법에 따른 배발생율은 처리구 I에서 92.2 %(83/90)으로 처리구II와 III의 62.4%(63/101)와 77.8%(144/185)에 비하여 높게 나타났으나 배반포 발달율은 처리구II와III에서 65.1%(41/63)와 50.0%(72/144)로 처리구 I의 30.1%(25/83)보다 높게 나타났다. 각 처리구에 따른 수정란 이식후 수태율은 처리구II와 III에서 공히 20%의 수태율을 나타낸 반면 처리구 I에서는 수태가 되지 않았다. 따라서 체세포 복제수정란의 생산에 있어서 배반포 발달율과 수태율을 높이기 위해서는 단순배양보다 공배양이 더 효과적인 것으로 사료되지만 이런 결과가 복제송아지 생산효율에 있어서도 효과적일지는 향후 더 많은 연구가 있어야 할 것으로 사료된다.

• Applied Microscopy
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• v.36 no.4
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• pp.299-305
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• 2006

Onuf's nucleus, which is located in the ventral horn, has been known to innervate the striated muscles of the urethral and anal sphincter muscles via the pudendal nerve Onuf's nuclei are resistant to pathologic condition such as poliovirus. The reason why the motor neurons in Onuf's nucleus are less degenerated is not certain until now. The present study aims at updating the microscopical characteristics including its location the Onuf's nucleus innervating the external urethral sphincter, and ultrastructures of the zinc-enriched (ZEN) terminals synaptically-contacting with Onuf's motor neurons in the rat spinal gray matter by using HRP tracing method and zinc selenium autometallography, respectively. Based on HRP tracing method, Onuf's nuclei were located adjacent lateral dendritic projections of the ventral horn. Their shape was almost round at lumbar level, but oval at sacral segment of spinal cord. In size, their somata were smaller than that of other motor nuclei. In AMG stained sections, Onuf's nuclei were innervated by highly concentrated ZEN terminals, and contained small and middle-sized ZEN, but totally void of large ZEN terminals. AMG silver grains were confined to presynaptic ZEN terminals against dendritic elements and somata of the Onuf's motor neurons. A majority of the ZEN terminals contained flattened synaptic vesicles and made symmetrical synaptic specializations.

• Journal of radiological science and technology
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• v.28 no.3
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• pp.249-254
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• 2005

This study investigated the structural changes of folliculus ovarii according to the dose of the X-rays when mice were exposed to X-rays from 6MeV LINAC. The minute structural changes of folliculus ovarii were observed through an electron microscope with high magnification. Nuclei and protoplasm of granular cells in growing folliculus ovarii abruptly underwent minute structural changes according to the increase of dose of X-rays. Cell residue, by-product of cell decease, neutrophil and macrophage around follicular antrum were observed. The minute structural changes in granular cells showed typical characteristics of apoptosis: the increase of electronic density due to nuclear condensation, fragmentation of nuclei, and atrophy of protoplasm. Necrosis of cells was identified, but it was not so remarkable. Macrophage scattered with apoptotic bodies.

• The Korean Journal of Zoology
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• v.26 no.4
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• pp.253-270
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• 1983

Electron microscopic degeneration studies were carried out on the rat's interpeduncular nucleus following lesion on both habenular nuclei to clarify the morphological nature of their presynaptic endings involved in the habenulo-interpeduncular pathway. Two distinct types of interpeduncular neurons were observed: 30% of large neurons and 70% of small ones. Four types of boutons with many characteristics could be differentiated; small spherical vesicle-containing boutons (40%), large spherical vesicle-containing boutons (18%), pleomorphic vesicle-containing boutons (33%), and elongated vesicle-containg boutons (9%). Vesicle containing dendrites were observed as dendro-dendritic synapses and axo-axonal synapses were also found, though rarely. Two to four days after the lesion on both habenular nuclei, the small spherical vesicle-containing boutons were found almost exclusively in the process of degeneration, while the other types were entirely free of degeneration. Therefore, it is inferred that habenural neurons terminate in the interpeduncular nucleus as small spherical vesicle-containing boutons. The neuronal origin of the other bouton types is discussed.