• Asian-Australasian Journal of Animal Sciences
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• 제32권5호
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• pp.629-636
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• 2019

Objective: The study was designed to determine the cytotoxic effect of gallic acid (GA), obtained by the hydrolysis of tannins, on mice TM4 Sertoli cells apoptosis. Methods: In the present study, non-tumorigenic mice TM4 Sertoli cells were treated with different concentrations of GA for 24 h. After treatment, cell viability was evaluated using WST-1, mitochondrial dysfunction, cells apoptosis and necrosis was detected using JC-1, Hoechst 33342 and propidium iodide staining. The expression levels of Cyclin B1, proliferating cell nuclear antigen (PCNA), Bcl-2-associated X protein (BAX), and Caspase-3 were also detected by quantitative real-time polymerase chain reaction and Western-blotting. Results: The results showed that 20 to $400{\mu}M$ GA inhibited viability of TM4 Sertoli cells in a dose-dependent manner. Treatment with $400{\mu}M$ GA significantly inhibited PCNA and Cyclin B1 expression, however up-regulated BAX and Caspase-3 expression, caused mitochondrial membrane depolarization, activated Caspase-3, and induced DNA damage, thus, markedly increased the numbers of dead cells. Conclusion: Our findings showed that GA could disrupt mitochondrial function and caused TM4 cells to undergo apoptosis and necrosis.

• 동의생리병리학회지
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• 제28권4호
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• pp.384-389
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• 2014

The purpose of this study is to investigate the modulatory effect of oroxylin A on hydrogen peroxide production in TM4 mouse testis sertoli cells induced by the synthetic analog of double-stranded RNA [polyinosinic-polycytidylic acid]. Hydrogen peroxide production was measured by dihydrorhodamine 123 (DHR) assay. Oroxylin A significantly inhibited the polyinosinic-polycytidylic acid (PIC)-induced production of hydrogen peroxide for 0.5, 2, 12, 18, and 24 hr incubation at the concentrations of 5, 10, 25, and $50{\mu}M$ in TM4 (P < 0.05) in dose dependent manner. These results suggest that oroxylin A has a protective effect against PIC-induced cellular toxicity with its inhibition of hydrogen peroxide production in PIC-induced sertoli cells.

• Toxicological Research
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• 제17권4호
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• pp.267-272
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• 2001

This study was carried out to assess the reproductive toxicity of 2-bromopropane (S-BP) using spermatogenesis stage classification and Sertoli cell indices (SCI).Vehicle control olive oil and 2-BP doses of 125, 250 and 500 mg/kg of body weight were injected in the interaperitoneum of 12 weeks male Sprague-Dawley rats for 28 days respectively of SCI on germ cells including the spermatogonia of stages II-III, Ⅵ,Ⅹ, XII, ⅩIII, and spermatocytes of stages VIII (preleptotene), Ⅹ (leptotene), XII (leptotene), V and Ⅵ (pachytene), and the round spermatids of stage Ⅵ. Considering the process of maturation depletion in spermatonesis, spermatogonia may be the primary target cells of 2-BP toxicity.

• 한국임상수의학회지
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• 제36권1호
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• pp.78-81
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• 2019

Some canine typical Sertoli cell tumors (SCT) induce signs of hyperestrinism. However, whether variant lipid-rich SCTs have signs of hyperestrinism remains largely unknown. In the present study, an 11-year-old male Pekingese dog showed significantly elevated serum estrogen and characteristic signs of hyperestrinism such as gynecomastia and alopecia. Cytological, radiological and ultrasound examinations found testicular mass, prostatitis with squamous metaplasia, and cystitis. Pathologically, the tumor lesions consisted mainly of lipid-rich tumor cells with signet-ring appearance, which were immunohistochemically positive for vimentin and anti-$M{\ddot{u}}llerian$ hormone. Based on the findings, a diagnosis of lipid-rich Sertoli cell tumor was established. In conclusion, a canine lipid-rich SCT induced signs of hyperestrinism and caused prostatitis via squamous metaplasia due to its excessive secretion of estrogen.

• 환경생물
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• 제17권3호
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• pp.285-292
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• 1999

플라스틱 제품의 가소제로 널리 사용되며, 최근 내분비 교란물질로 알려져 있는 di-(2-ethylhexyl)phthalate(DEHP)를 흰쥐에 15일 동안 구강 투여(1g/kg/day, 2g/kg/day, 3g/kg/day)한 후, 정자형성과정에 연관된 정소의 기능과 구조에 미치는 영향을 조사하였다. DEHP 처리군에서는 대조군에 비하여 체중 증가율이 감소하였을 뿐만 아니라 정소의 무게도 감소하였다. 또한 세정관의 직경이 고농도군으로 갈수록 작아지는 경향을 보였으며 세정관내의 세포층이 감소하는 현상이 나타났다. 미세구조의 변화를 관찰한 결과, 실험군은 세정관내 세포사이 공간이 증가하였으며 정원세포와 정모세포의 수가 감소하였고, 세포질내 공포가 증가하는 것이 관찰되었다. 또한 세정관내 Sertoli 세포의 전체 세포질 양이 감소하는 경향을 보였고, 정원세포의 경우 핵막 이중층이 분리되는 현상을 보였다. 특히 고농도군의 세정관내에서는 Sertoli 세포 이외의 세포는 거의 관찰되지 않았으며, 세포사이 공간과 공포들이 상당히 증가하였다. Sertoli 세포의 핵막은 심하게 함입된 형태를 나타냈으며 이질염색질이 증가하여 염색질의 덩어리를 이루고 있었고, Sertoli 세포들을 지지하는 기저판은 심하게 굴곡된 형태를 보였다. Leydig 세포의 미세구조에 있어서도 실험군은 대조군과 비교하여 현저한 차이를 보였다. 세포질내의 활면소포체와 핵막 이중층이 심하게 팽대되는 경향을 보였으며 핵내에 이질염색질이 상당히 증가하고 세포질내 리소솜이 증가하는 특징을 보였다.또한 혈청내 테스토스테론 함량에 있어서도 실험군은 현저히 낮은 수치를 나타냈다. 결론적으로, DEHP는 정소의 발달을 농도의존적으로 저해하고, Leydig 세포의 테스토스테론 합성기능을 저해하며, 이어서 세정관내부의 Sertoli 세포의 구조와 기능을 손상시켜 생식세포들의 괴사를 유도하는 과정을 통해 일련의 정자형성과정을 억제하는 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제27권11호
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• pp.1554-1561
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• 2014

Present study analyzed the changes in peripheral blood testosterone concentrations and testicular cytogram in relation to age and semen quality in crossbred males. Three different age groups of crossbred males viz. bull calves (6 months, n = 5), young bulls (15 months, n = 5) and adult bulls (4 to 6 years, n = 8) were utilized for the study. Testicular fine needle aspiration cytology technique was used to quantify testicular cytology and their indices. Peripheral blood testosterone concentrations were measured using enzyme-linked immunosorbent assay method. Semen samples collected from adult bulls were microscopically evaluated for quality parameters. Mean peripheral blood testosterone concentrations in bull calves, young bulls and adult bulls were $2.28{\pm}0.09ng/mL$, $1.42{\pm}0.22ng/mL$ and $5.66{\pm}1.08ng/mL$ respectively, and that in adult bulls were significantly different (p<0.01) from young bulls and bull calves. There was no significant difference between the proportion of different testicular cells in bull calves and young bulls. Between young and adult bulls, significant differences (p<0.01) were observed in the proportion of spermatocytes, spermatozoa, and sperm: Sertoli cell ratio. The proportions of Sertoli cells showed a significant difference (p<0.01) between the three age groups. The number of primary spermatocytes had a positive correlation with peripheral blood testosterone concentrations in bull calves (r = 0.719, p<0.01). Number of Sertoli cells per 100 germ cells was negatively correlated with blood testosterone concentration in young bulls (r = -0.713, p<0.01). Among different semen parameters in adult bulls, ejaculate volume (r = 0.790, p<0.05) had positive relationship, and sperm motility had significant negative correlation (r = -0.711, p<0.05) with testosterone concentrations. The number of Sertoli cells and Sertoli cell index had a positive correlation with various semen quality parameters (p<0.001). Results of the present study conclude that number of Sertoli cells and Sertoli cell index are good indicators of semen quality, but peripheral blood testosterone concentrations may not have a direct relationship with various seminal attributes in crossbred bulls.

• 한국발생생물학회지:발생과생식
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• 제4권1호
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• pp.37-43
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• 2000

생쥐 정소에서 밀착결합단백질의 일종인 zonular occludens-1 (ZO-1)의 발현을 조사하였다. RT-PCR결과 ZO-1의 2가지 isoform인 ZO-1$\alpha$＋, ZO-1$\alpha$-의 발현을 확인하였다. 생쥐 신생 및 성체의 정소에서 분자량 225 및 2001 KDa의 2종의 ZO-1의 단백질항원의 발현을 확인되어 RT-PCR의 결과와 일치하였다. ZO-l$\alpha$＋에 대한 ZO-l$\alpha$-의 상대적 발현량은 성숙에 따라 증가하였다. 2종의 ZO-1항원을 동시에 인식하는 항체를 사용한 면역염색을 통해 세정관 외곽의 Sertoli세포 사이의 접촉부위 및 Sertoli 세포와 생식세포 접촉부위에서 ZO-1의 존재를 확인하였다. ZO-1은 세정관내 세포들 사이의 결합부위 및 세포질에서 공통적으로 발현되지만 성숙에 따라 Sertoli 세포의 결합부위에서 강한 신호가 검출되었다. 2종의 ZO-1 항원의 상대적 발현량의 변화 및 세정관 외곽의 분포의 강화는 기능적 혈액정소장벽의 출현 및 정자형성의 진행과 관련된 것으로 사료된다.

• 한국수정란이식학회지
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• 제33권3호
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• pp.177-183
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• 2018

생체 조직 내에서 표지인자의 발견은 해당 세포의 특성과 기능을 이해하는 데 매우 중요하다. 이전의 연구에서 본 연구진은 돼지정원줄기세포에서 특이적으로 IGFBP 3가 발현되어 표지인자로의 가능성을 보고한 바 있다. 본 연구에서는 IGFBP 3이외의 다른 family member가 정원줄기세포에서 특이적으로 발현하는 지와, 이의 발현을 조절하는 PAPP-A의 조직학적인 측면에서의 발현 양상을 5일령 돼지 정소에서 확인하였다. 그 결과 IGFBP 1, 2, 3, 4, 6의 발현은 정소 전체에서 발현되는 수준보다 돼지 정원줄기세포에서 더 높은 수준에서 발현하고 있음을 확인하였다. PAPP-A는 sertoli cell에서 특이적으로 발현하며, 정원줄기세포에서는 발현하지 않는 것을 PGP9.5와의 동시-조직염색으로 확인하였다. 이러한 결과는 Sertoli cell에서 발현하는 PAPP-A 단백질은 미성숙 돼지 정소에서 IGFBP family를 통해 정소 세포의 발달과 분화를 조절할 것으로 판단된다.

• Applied Microscopy
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• 제16권2호
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• pp.75-91
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• 1986

Differentiation of the rat testis was studied by light and electron microscope from the fetal stage up to the newborn or adult stage. The purpose of the present study is to investigate the ultrastructural changes of seminiferous tubules and interstitial tissue during the developmental process. The results were as follows: the seminiferous tubule diameter began to increase from birth and was fully developed at 30 to 40 days of age through intratubular cell proliferations. Basement membrane and myoid cells lining the seminiferous tubules were differentiated at 17 days gestation. At the fetal stage, seminiferous tubules were primarily composed of Sertoli cells and the differentiation of Sertoli and germ cells progressed from the newborn stage. Spermatids and immature spermatozoa are appeared at 40 days of age, so from this time, spermatogenesis occurred actively until the adult stage. Sertoli cells aided germ cell differentiation and phagocytosed the parts of the spermatid cytoplasm. Leydig ce]] development follows a biphasic pattern: a fetal phase and then an adult phase from 20 days of age. In conclusion, the rat testis is already developed to some extent by the fetal stage and is functional after 50 days of age. Therefore, these findings indicate that differentiation of Sertoli and Leydig cells precedes the onset of spermatogenesis.

• 대한수의학회지
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• 제36권3호
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• pp.531-539
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• 1996

The lectin-binding patterns in the testis of the sexually matured Jindo dog were investigated to study the distribution of glycoconjugates in the seminiferous tubule under light and transmission electron microscopy. Positive reactions to Wheat germ agglutinin(WGA) and Dolichos biflorus agglutinin (DBA) were observed in the Sertoli cell and in the residual body of spermatid with a stronger reaction in the Sertoli cell to the lectins than in the residual body. Strong reactions to Soybean agglutinin(SBA) and Peanut agglutinin(PNA) were observed in the acrosome vesicles of the Golgi- and cap-phase spermatid, while a moderate reaction was observed in the acrosome-phase, maturation-phase spermatid and the residual body. The acrosome area of the spermatid reacted intensively to Griffonia simplicifolia agglutinin( GS-I) when the cell was in the acrosome-phase and maturation-phase, and the same reaction to the GS-I was observed in the residual body. However, the seminiferous tubule did not react to Ulex europeus agglutinin I(UEA-I). The gold-labelling of the Sertoli cells with DBA resulted in positive reactions of the Sertoli cell column and processes when observed under the electron microscopy, while the Golgi-, cap- and acrosome-phase spermatids reacted positively to SBA in the peripheral low-dense area of the acrosome vesicle of spermatid. Based on these results, we concluded that differences in the lectin-binding pattern of the seminiferous tubules were recognized in the Jindo dog compared to other animals.