• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.48-48
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• 2003

본 실험은 spermatogonia 단계에 발현하는 유전자를 찾기 위하여 suppression subtractive hybridization를 수행하였다. 기존에 mouse에서는 spermatogonia 특이적인 유전자들이 밝혀져 있기 때문에 pig에 특이적인 유전자를 찾기 위하여 pig 250days testis와 pig 60days testis를 재료로 하여 실험하였다. SSH를 통하여 254days testis에 특이적으로 발현되는 후보유전자를 7개 찾았고 25days testis와 60days testis 의 Northern blot을 통하여 25days에 과발현하고 60days에 발현의 양이 대폭 줄어드는 spermatogonia 유전자로 생각되는 후보유전자 2개를 선택하여 pig tissue northern blot, genomic DNA southern blot, RT-PCR 그리고 In-situ hybridization을 수행하였다. Tissue northern blot과 RT-PCR을 통하여 후보자 1번은 간과 폐, 난소, 정소에서 발현하고, 후보유전자 15번은 난소와 정소에서만 특이적으로 발현함을 알았다. DNA sequence analysis와 NCBI Blast search를 통하여 후보자 1번은 다른 종에서 밝혀진 유전자였고 후보유전자 15번은 어느 종에서도 밝혀지지 않은 새로운 유전자였다. Degenerated primer를 통하여 후보자 1번의 pig full sequence를 밝히고 NCBI에 등록하였다. 그리고 In-situ hybridization을 통하여 후보유전자득이 20일째 testis의 Leydic cell에서 많이 발현되고 adult testis에서는 발현이 감소하는 결과를 얻었다. 이것으로 보아 위의 두 후보유전자는 spermatogonia에 직접 관련된 유전자이기 보다는 spermatogonia의 발달에 영향을 주는 leydic cell 특이발현을 가진 유전자로 사료되어진다.

• 대한수의학회지
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• 제26권2호
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• pp.201-210
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• 1986

The cycle of the seminiferous epithelia in the testis of mature Korean native cattle was divided into twelve stages, using criteria the morphological changes in the acrosomic system and the nuclei of developing spematids and germ cells. The results were summarized as follows; 1. The minimum number of tripe A spermatogonia were the average of 1.8 in both at stages I and VI, while maximum numbers were the average of 4.2 at stage XII. Some type A spermatogonia divided at stage XII to produce the type intermediate(IN) spermatogonia at following stage I. The intermediate type spermatogonia divided at stage IV to produce the type B spermatogonia at stage V. 2. The type B spermatogonia divided at stage VII to produce the preleptotene primary spermatocytes at stage XII. The pachytene primary spermatocytes divided at stage XI to produce the secondary spermatocytes at stage VII. The secondary spermatocytes observed at stag XII divided to give rise to the round spermatids at following stage I. Each numbers of the first spermatocytes and of spermatids were almost constant, respectively, through the cycle of the seminiferous epitherium. 3. The relative frequencies of each stage among stages I to XII of the cycle of the seminiferous epithelia were 6.1, 3.7, 5.2, 7.8, 2.2. 3.3, 13.8, 18.4, 11.8, 7.2, 18.1% and 2.4%, respectively.

• 한국수정란이식학회지
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• 제20권1호
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• pp.35-41
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• 2005

본 연구는 가축유전자원의 효율적 보존을 위해 정조세포를 줄기세포 형태로 장기보관하면서 필요에 따라 증식, 분화를 통해 가축의 복원에 활용하기 위한 연구의 일부로 진행되었다. 정조세포를 분리하여 배양한 결과 배양온도는 다른 세포들과는 달리 $32^{\circ}C$에 세포분열이 활발하였으며, TCM199에 $10\%$ FCS를 첨가한 배양액과 세르톨리세포 공배양으로 정조세포의 배양을 지지하였다. 40일령이 지나면서 정조세포 콜로니 즉 germline stem cells를 형성하였으며, 일부에서는 외형상 ES-like cells를 형성하거나, 세정관 형태로 정조세포들이 재구성되었다. 40일령까지 배양한 상태에서는 정조세포의 정모세포나 정자세포로 분화하는 징후를 관찰할 수 없었으며, 추후 이들 세포로 분화를 유기하는 실험이 진행되어야 할 것이다.

• 대한수의학회지
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• 제32권3호
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• pp.281-293
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• 1992

Classification of the cycle of seminiferous epithelia into 12 stages by the morphological changes in acrosomal system and evaluation of the relative frequency of stages and the cell association were histologically performed in the mature Korean native Jin-do dogs. The results were summarized as follows; 1. The minimum number of type A spermatogonia averaged 1.01 at stages I, while maximum number averaged 2.47 at stages XII. Some type A spermatogonia divided at stage XII to produce the type intermediate(IN) spermatogonia at the following stage I. The type IN spermatogonia divided at stage IV to produce the type B spermatogonia at stage V. 2. The type B spermatogonia divided at stage VI to produce the preleptotene primary spermatocytes at stage VII. The secondary spermatocytes observed at stage XII. The secondary spermatocytes observed at stage XII divided to give rise to the round spermatids at the following stage I. The numbers of the first spermatocytes and spermatids were almost constant, respectively, through all the cycles of seminiferous epithelium. 3. The acrosomal vesicle was invaginated to occupy one third to half of spermatid nucleus at the cap phase, which was different from that of rodent and ruminant spermatid nuclei. 4. The relative frequencies of stages I to XII of seminiferous epithelia cycle were 10.34, 4.84, 5.03, 8.22, 10.86, 6.63, 6.42, 18.88, 10.17, 6.18, 7.62% and 4.81%, respectively.

• 한국발생생물학회지:발생과생식
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• 제19권1호
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• pp.1-10
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• 2015

The purpose of the present study was to examine the seminiferous epithelium cycle of Bombina orientalis using a light microscope. The cycle was divided into a total of 10 stages, according to the morphological characteristics of the cells. The spermatogenetic cells included primary spermatogonia, secondary spermatogonia, primary spermatocytes, secondary spermatocytes, spermatid and sperm. At stage I, the primary spermatogonia was located closer to basal lamina of the seminiferous tubule without spermatocyst formations. Especially at the stage II, the secondary spermatogonia were located in the spermatocyst. The primary and secondary spermatocytes were found from stages III to VI. The secondary spermatocytes were smaller in size than the primary spermatocytes, but they had thicker nucleoplasm and smaller nuclei. The round-shaped, early sperm cells were formed in stage VII, and further divided at stage VIII to have more concentrated nucleoplasm before division to matured sperm cells. At stage X, the matured sperm cells emerged from the spermatocyst. Considering the above results, this study presented the special characteristics in the generation and type of sperm formation. The germ cell formation occurred in various stages, like the perspectives of Franca et al (1999), ultimately, providing taxonomically useful information.

• 대한수의학회지
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• 제33권1호
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• pp.23-36
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• 1993

To investigate the cycle and relative frequences and the fine structure of seminiferous epithelia in mature Jindo dogs, histologic study was performed. The results obtained were summarized as follows; 1. Type A spermatogonia appeared approximately 1.6 times as many at stage II as compared to stage I while type In spermatogonia appeared small amount in stage III, IV and V. type B spermatogonia were found during the stage VI to VIII, though not detectable during stage I to V. The type B spermatogonia divided at stage VII to produce the preleptotene primary spermatocytes at stage VIII. The number of primary spermatocytes of the leptotene phase markedly increased during stage I to II, and the primary spermatocytes of the pachytene phase were shown the least in number at stage IV. The secondary spermatocytes could be seen only at stage IV. 2. The relative frequencies of each stage from stages I to VIII of the cycle of seminiferous epithelia were 31.6, 11.9, 10.0, 3.2, 8.2, 10.1, 11.7 and 13.2% respectively. 3. On electron microscopic observations, acrosomal vesicle of spermatids appeared larger though the bulk of germ cells were the morphologically same as those of the other animal species. Thread line structures light microscopically observed in the cytoplasm of Sertoli cell were the longitudinal orientation of mitochondria.

• Radiation Oncology Journal
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• 제8권1호
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• pp.17-27
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• 1990

온열요법이 X-선조사에 미치는 영향을 연구하기 위하여 저자는 100 watt, 2450 MHz의 마이크로파 온열기구를 제작하여 백서 60마리를 대상으로 고환에 온열요법 ($43^{\circ}C$, 15분), X-선 조사($2\;Gy\~8\;Gy$ 단독 혹은 병행하여 조직학적 변화를 관찰하여 다음과 같은 결론을 얻었다. 1. 2450 MHz 마이크로파 온열요법 단독 시행하여 조직학적 변화는 관찰되지 않았다. 2. X-선 조사후 초기변화는 정조세포의 변성을 2 Gy에서 나타나기 시작하였으며 괴사는 6 Gy 후 심하게 보였고 8 Gy 후 완전소실되었다. 3. X-선 조사와 온열요법 병행하여 2 Gy후 정조세포의 괴사와 소실이 나타나기 시작하였으며 6 Gy 후 완전소실되어 열증강율은 1.3 이었다. 4. X-선조사, 온열요법 단독 및 병행후 추적기간 15일군과 30일군의 차이는 관찰할 수 없었다.

• Animal cells and systems
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• 제8권3호
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• pp.197-203
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• 2004

CD30 is a member of tumor necrosis factor receptor (TNFR) superfamily and has pleiotropic functions including cell activation, proliferation, differentiation, and death, depending on cell types and stage of differentiation. Although CD30 expression has been described mainly in hematopoietic tissues, several types of nonhematopoietic tumors including embryonic carcinoma and germ-cell tumors express CD30. We examined CD30 distribution in the testis and epididymis from wild type and CD30-deficient mice. In the testis, spermatogonia, spermatocytes and Sertoli cells expressed CD30, but not in spermatids. Spermatogonia and spermatocytes near the basement membrane strongly reacted to anti-CD30. In the epididymis, CD30 expression was exclusively observed in luminal epithelia and some interstitial cells. Taken together, these results show a spatio-temporal regulation of CD30 expression in mouse testis and epididymis and suggest a possible role of CD30 in spermatogonia and spermatocytes.

• 생명과학회지
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• 제15권3호
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• pp.387-396
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• 2005

The aims of the study were to establish a short-term screening test for detecting testicular toxicity of chemicals in rats and to determine whether a 2-week administration period is sufficient to detect testicular toxicity of 2-bromopropane (2-BP) as an example. Male Sprague-Dawley rats were subcutaneously administered with 1000 mg/kg/day of 2-BP or its vehicle for 2 weeks. Ten male rats each were sacrificed on days 3, 7 and 14 after the initiation of treatment. Parameters of testicular toxicity included genital organ weights, testicular sperm head counts, epididymal sperm counts, motility and morphology, and qualitative and quantitative histopathologic examinations. The early histopathological changes observed on day 3 of treatment included degeneration of spermatogonia and spermatocytes, multinuclear giant cells, mature spermatid retention, vacuolization of Sertoli cells, and decreased number of spermatogonia in stages II and V. On day 7 of treatment, atrophy of seminiferous tubules, exfoliation of germ cells, degeneration of spermatogonia and spermatocytes, multinuclear giant cells, mature spermatid retention, vacuolization of Sertoli cells, decreased number of spermatogonia in stages II and V, and decreased number of spermatocytes in stages VII and XII. On day 14 after treatment, a significant decrease in the weights of testes and seminal vesicles was found. Atrophy of seminiferous tubules, exfoliation of germ cells, degeneration of spermatogonia and spermatocytes, mature spermatid retention, vacuolization of Sertoli cells, decreased number of spermatogonia in stages II and V, and decreased number of spermatocytes in all spermatogenic stages were also observed. In addition, a slight non-significant decrease in testicular sperm head counts, daily sperm production rate and epididymal sperm counts was found. The results showed that 2 weeks of treatment is sufficient to detect the adverse effects of 2-BP on male reproductive organs. It is considered that the short-term testicular toxicity study established in this study can be a useful tool for screening the testicular toxic potential of new drug candidates in rats.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2002년도 국제심포지엄
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• pp.63-65
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• 2002

Identification of spermatogonial stem cell-specific surface molecules is important in understanding the molecular mechanisms underlying the maintenance and differentiation of these cells. We have found that spermatogonia from busulfan treated mice expressed an autoantigen that distinguishes between undifferentiated and differentiated spermatogonia. Four to six weeks after busulfan treatment, germ cells located in the basal compartment of seminiferous epithelium show isotype-specific IgG deposits that form due to autoimmunity. Before busulfan treatment, the level of testicular IgG was very low but IgG levels began to increase after week 4 and peaked at week 6. When cells from the busulfan treated testis were analyzed using laser scanning cytomeoy (LSC), the frequency of cells positive for IgG deposits, 6-integrin, and 1-integrin were 16.5${\pm}$3.8%, 11.8${\pm}$2.6%, and 9.0${\pm}$ 1.4%, respectively. Immunofluorescent staining suggested that most, if not all of the cells with IgG-deposits isolated from a laminin-coated dish, were also positive for a spermatogonial stem cell marker \ulcorner6-integrins as well as for a germ cell-specific marker TRA 98. We determined serum and intratesticular IgG levels and the soundness of seminiferous tubule basement membrane from busulfan treated mice using electron microscopy, in order to study the mechanism responsible for IgG deposits in spermatogonia. We found that the basement membranes of seminiferous tubules from busulfan treated mice were severely impaired when compared to those of normal adult, neonates and w/wv mice. Furthermore, new blood cells were observed in the surface of the damaged basement membrane along the seminiferous tubules. These results suggest that the IgG in spermatogonial stem cells accumulates from circulating blood through the impaired basement membranes induced by busulfan treatment. Taken together, our study suggests that IgG can be used as a new marker for undifferentiated spermatogonia cells.