• 한국동물생명공학회지
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• 제38권4호
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• pp.291-299
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• 2023

Background: Leydig cells, crucial for testosterone production, express ion channels like ANO1 that influence hormone secretion. This study investigates the expression and role of the Tandem of P domains in a weak inward rectifying K⁺ channel-related Acid-Sensitive K⁺-1 (TASK-1) channel in these cells, exploring its impact on testicular function and steroidogenesis. Methods: TASK-1 expression in Leydig cells was confirmed using immunostaining, while RT-PCR and Western Blot (WB) validated its expression in the TM3 Leydig cell line. The effect of a TASK-1 channel blocker on cell viability was assessed through live/dead staining and MTT assays. Additionally, the blocker's effect on testosterone secretion was evaluated by measuring testosterone levels. Results: Immunohistochemical analysis revealed a predominant presence of TASK-1, along with c-Kit and ANO-1, in Leydig cells adjacent to seminiferous tubules and also in Sertoli and spermatogenic cells. Expression levels of TASK-1 mRNA and protein were significantly higher in TM3 Leydig cells compared to TM4 Sertoli cells. In addition, blocking TASK-1 in TM3 cells with ML365 induced cell death but did not affect LH-induced testosterone secretion. Conclusions: These findings suggest that TASK-1 in Leydig cells is crucial for their viability and proliferation, highlighting its potential importance in testicular physiology.

• 대한본초학회지
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• 제36권5호
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• pp.117-123
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• 2021

Objectives : The aim of this study is to investigate the effect of wogonin on the production of hydrogen peroxide in polyinosinic:polycytidylic acid (poly I:C)-stimulated TM4 mouse sertoli cells. Methods : TM4 were treated with poly I:C (50 ug/mL) and wogonin at concentrations of 5, 10, 25, and 50 µM for 30 min, 2 hr, 12 hr, 18 hr, and 24 hr. The production of intracellular hydrogen peroxide was measured by dihydrorhodamine 123 assay. Results : For 30 min, 2 hr, 12 hr, 18 hr, and 24 hr treatment, wogonin significantly inhibited intracellular hydrogen peroxide productions of TM4 at the concentration of 5, 10, 25, and 50 µM (p<0.05). In details, production of hydrogen peroxide in poly I:C-stimulated TM4 treated for 30 min with wogonin at concentrations of 5, 10, 25, and 50 µM was 95.67%, 92.69%, 92.05%, and 91.97% of the control group treated with poly I:C only, respectively; the production of hydrogen peroxide for 2 hr was 94.44%, 94.41%, 93%, and 92.98%, respectively; production of hydrogen peroxide for 12 hr was 96.78%, 95.32%, 94.33%, and 93.17%, respectively; production of hydrogen peroxide for 18 hr was 94.7%, 93.4%, 93.38%, and 93.35%, respectively; and production of hydrogen peroxide for 24 hr was 95.75%, 94.77%, 94.58%, and 92.8%, respectively. Conclusions : Wogonin might have anti-viral property related with its inhibition of intracellular hydrogen peroxide production in poly I:C-stimulated TM4 cells.

• Archives of Pharmacal Research
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• 제24권4호
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• pp.360-366
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• 2001

Gangliosides are ubiquitous membrane components in mammalian cells and are suggested to play important roles in various functions such as cell-cell interaction, adhesion, cell differentiation, growth control and signaling. Among all ganglio-series gangliosides, GM3 has the simplest carbohydrate structure, and has been shown as a major gangliosides, in male reproductive system. To study GM3 distribution in the seminiferous tubule and epididymis, frozen sections were stained with specific monoclonal antibody (MAb) against ganglioside GM3. In the seminiferous tubule of testis, pachytene spermatocytes and spermmatids expressed ganglioside GM3, but not in spermatogonia and sertoli cells. Spermatogonia and sertoli cells near the basement membrane were negatively reacted to anti-GM3. In the epididymis, GM3 was expressed only in some interstitial cells. Taken togethers, these results suggest that the expression of ganglioside GM3 in rat seminiferous tubule and epididymis is spatio-temporally regu lated during spermatogenesis.

• Applied Microscopy
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• 제38권3호
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• pp.205-212
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• 2008

6-aminonicotinamide (6-AN)의 햄스터 정소에 미치는 영향을 확인하기 위해 실험군에는 체중 kg 당 10 mg의 6-AN을, 대조군에는 동량의 생리식염수를 격일로 복강 투여한 후 정소의 변화를 광학 및 투과전자현미경을 이용하여 관찰하였다. 최초 6-AN 투여 당시의 체중에 비해 7회 투여군부터 유의하게 체중이 감소하였으며, 정소 중량의 감소는 5회 투여시 크게 감소하여 이후 비슷한 양상을 보였다. 정세관상피의 퇴행변화는 5회 투여군부터 나타나기 시작하여 9회 투여군부터는 대부분의 정세관이 심하게 손상을 받은 것으로 나타났다. 손상을 받은 정세관에서 정세관상피를 이루는 정자발생세포 및 지지세포 모두 심한 공포화에 따른 세포의 파괴를 뚜렷하게 관찰할 수 있었으며, 다핵거대세포가 출현하였다. 사이질조직의 부종은 관찰할 수 없었으며, 사이질세포 역시 비교적 온전하게 보존되어 있었다. 따라서 6-AN은 햄스터 정소에서 정자발생세포, 지지세포 등에는 영향을 미치나 사이질세포에는 영향을 주지 않는 것으로 사료된다.

• Clinical and Experimental Reproductive Medicine
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• 제26권1호
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• pp.103-109
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• 1999

Objective: Heat shock protein 70-2 (Hsp70-2) gene knockout mice are found to have premeiotic arrest at the primary spermatocyte stage with a complete absence of spermatids and spermatozoa. This observation led to the hypothesis that hspA2 may be disrupted in human testes with abnormal spermatogenesis. To test this hypothesis, we studied the mRNA expression of hspA2 in infertile men with azoospermia. Design: The mRNA expression were analyzed by competitive RT-PCR among testes with normal spermatogenesis, pachytene spermatocyte arrest, and sertoli-cell only syndrome. Materials and methods: Testicular biopsy was performed in men with azoospermia (n=15). Specimens were subdivided into three groups: (group 1) normal spermatogenesis (n=5), (group 2) spermatocyte arrest (n=5), (group 3) Sertoli-cell only syndrome (n=5). Total RNA was extracted by Trizol reagent. Total extracted RNA was reverse transcribed into cDNA and amplified by PCR using specific primers for hspA2 target cDNAs. A competitive cDNA fragment was constructed by deleting a defined fragment from the target cDNA sequence, and then coamplified with the target cDNA for competitive PCR. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was used as an internal control. Results: On Competitive RT-PCR analyses for hspA2 mRNA, significant amount of hspA2 expression was observed in group 1, whereas a constitutively low level of hspA2 was expressed in groups 2 and 3. Conclusion(s): The study demonstrates that the hspA2 gene expression is down-regulated in human testes with abnormal spermatogenesis, which in turn suggests that hspA2 gene may play a specific role during meiosis in human testes.

• Toxicological Research
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• 제7권1호
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• pp.83-92
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• 1991

Complexities of testis structure and function are emphasized in morphometrical and genotoxic evaluation by statistical analysis. F-344 rats were treated with azinphos methyl, cyclophosphomide, and dichlorvos. And Brdu was injected with intrapertionially before sacrifice. The existence and degree of DNA damage were measured by Brdu labeling index which represented relative amount of Brdu incorporated in DNA, morphometric change was evaluated by the relative length of tubular diameter in circular seminiferous tubules and the number of spermatogonia per Sertoli cell in stage IX seminiferous tubules.

• 한국수의병리학회:학술대회논문집
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• 한국수의병리학회 2005년도 Proceedings of The 2nd Asian Society of Veterinary Pathology Symposium(Vol.2) and 2005 Annual Meeting of The Korean Society of Veterinary Pathology(Vol.9)
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• pp.76-76
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• 2005

• 대한수의학회지
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• 제40권2호
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• pp.361-371
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• 2000

최근 2-bromopropane(2-BP)이 사람과 실험동물에서 정소독성을 유발한다고 보고된 바 있다. 그러나 수컷 생식기계에 있어서 2-BP의 지연효과에 대해서는 세부적으로 조사된 바가 없다. 본 연구는 Sprague-Dawley 랫드에서 2-BP의 정소독성과 정자발생의 회복을 조사하기 위하여 수행하였다. 5주령의 수컷 랫드에게 2-BP를 1,000mg/kg 용량으로 4주간 반복투여하였고, 투여시작후 1, 2, 3, 4 및 12주째에 부검하였다. 정소독성의 평가는 병리조직학적인 질적평가와 생식기관 중량, 정자두부수 및 재생지수 등의 양적평가로 수행하였다. 시험결과 2-BP를 투여한 랫드에서는 체중과 정소 및 정소상체 중량이 대조군에 비해 시간의존적인 방식으로 억제 또는 감소하였다. 병리조직검사에서는 투여 1주째에 stage I~IV에서 정조세포와 stage VII~IX에서 세사전기 및 세사기의 정모세포가 현저하게 소실되었다. 정조세포는 투여 2주째에 모든 stage에서 광범위하게 소실되었으며, 정자발생주기가 진행됨에 따라 2, 3 및 4주째에는 접합기 정모세포, 비후기 정모세포 및 원형 정자세포가 전구세포의 결손에 의해 점진적으로 소실되었다. 지지세포의 기능적 이상을 암시하는 지지세포의 공포화와 정자세포 저류는 상기한 모든 시기에서 관찰되었다. 8주 회복후인 12주째에는 대부분의 곡세정관이 심하게 위축되어 지지세포만 관찰되었으며, 간질조직에서는 간질세포의 과형성이 인정되었다. 또한 2-BP에 의해 유발된 정소의 손상이 비가역적임을 암시해주는 정자두부와 재생지수의 현저한 감소가 관찰되었다. 상기결과는 랫드의 2-BP를 1,000mg/kg의 용량으로 4주간 반복투여하면 정조세포의 결손에 의해 점진적으로 생식세포가 감소하고 이로 인하여 장기적인 정소위축이 유발된다는 것을 암시해준다.

• 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.

• 한국수의병리학회지
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• 제2권1호
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• pp.47-52
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• 1998

The incidence of spontaneous testicular atrophy and its morphological changes during stage-specific spermatogenesis were investigated in male Sprague-Dawley rats at 10, 19, and 32 weeks of age. The incidence of testicular atrophy was 0.2%(2/90) 7.9%(9/114) and 10%(4/40) in 4, 13 and 26 weeks respectively. The epididymis with testicular atrophy had low sperm density. In the minimally affected tests scattered tubules showed complete depletion of germ cells without stage specificity. Testes with moderate to severe testicular atrophy showed seminiferous tubules lined with dense Sertoli cell population. While Leydig cells in the interstitium appeared hypertrophy they were immunohistochemically negative for proliferating cell nuclear antigen a marker of cell proliferation.