• 대한한방부인과학회지
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• 제32권1호
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• pp.1-14
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• 2019

Objectives: This study was designed to investigate the effect of Bombycis corpus on reproductive dysfunction caused by aging. Methods: The experimental group was divided into three groups: a control group consisting of 8-week-old male ICR mice without any treatment, An aging-elicited group (AE group) consisting of 50-week-old ICR male mice without any treatment, and a Bombycis corpus treatment group (BC group) consisting of 50-week-old ICR male mice with treatment Bombycis corpus extract (0.78 g/kg/day) for 6 months. After 6 months, histochemistry and immunohistochemistry of the testis were performed to investigate the effects of Bombycis corpus on the reproductive dysfunction caused by aging. Results: In the first step, Bombycis corpus increased spermatogenesis and distribution of sertoli cells in the seminiferous tubule, increased BrdU positive reaction in the spermatogonium at the basal part of the seminiferous tubule, and decreased the apoptosis of Sertoli cells in the seminiferous tubule. In addition, Bombycis corpus increased AR positive in Sertoli cells and $17{\beta}-HSD$ positive in leydig cells. Finally, Bombycis corpus decreased 8-OHdG positivite reaction in the spermatids of the seminiferous lumen, caspase-3 positivity in leydig cells, and HDAC1 positivite reaction in sertoli cells. Conclusions: These results suggest that Bombycis corpus increases spermatogenesis, decreases apoptosis of leydig cells and Sertoli cells, increases the production and action of testosterone in the testis, and inhibits DNA damages and DNA transcripts decrease in the testis, Thereby improving reproductive dysfunction caused by aging.

• 대한방사선기술학회지:방사선기술과학
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• 제18권1호
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• pp.97-110
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• 1995

This study was undertaken to investigate ultrastructural changes according to the radiosensitivity in the spermatogenic cells and Sertoli cell of the seminiferous tubules in Korean native pheasants. During spermatogenetic period, testes were collected from male adult Korean native pheasant and they were used as experimental and control birds. The experimental group was divided into a single-dose whole body irradiation group (400, 600, 800 and 1,000 rads) and a split-dose whole body irradiation groups(400/2, 600/2, 800/2 and 1,000/2 rads). The experimental birds were sacrificed at 24 and 72 hrs after irradiation and the control pheasants were sacrificed at the same time. Ultrastructural changes of Sertoli cells and spermatogonia were investigated by ultrathin section with electron microscope. The results obtained are summarized as follows; 1. The apoptosis was observed after 72 hrs group of the single-dose irradiation of 400 rads. 2. The cytoplasmic organelles of spermatogonia were severely damaged more than that of sertoli cell in 72 hours group of split-dose irradiation of 800 rads. 3. The cytoplasmic organelles of Sertoli cell were severely damaged except the nuclear membrane of Sertoli cells in 72 hrs group of split-dose irradiation of 1,000 rads.

• 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.

• 한국발생생물학회지:발생과생식
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• 제25권1호
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• pp.15-24
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• 2021

Benzo[a]pyrene (B[a]P) is a potent carcinogen and is classified as an endocrine-disrupting chemical. In mammalian testes, Sertoli cells support spermatogenesis. Therefore, if these cells are negatively affected by exposure to xenotoxic chemicals, spermatogenesis can be seriously disrupted. In this context, we evaluated whether mouse testicular TM4 Sertoli cells are susceptible to the induction of cytotoxicity-mediated cell death after exposure to B[a] P in vitro. In the present study, while B[a]P and B[a]P-7,8-diol were not able to induce cell death, exposure to BPDE resulted in cell death. BPDE-induced cell death is accompanied by the activation of caspase-3 and caspase-7. Depolarization of the mitochondrial membrane and cytochrome c release from mitochondria were observed in benzo[a]pyrene-7,8-diol-9,10-epoxide (BPDE)-treated cells. These results indicate that TM4 cells are susceptible to apoptosis in a caspase-dependent manner. Western blot and reverse transcription-polymerase chain reaction (RT-PCR) analyses showed that aryl hydrocarbon receptor (AhR) expression was almost undetectable in TM4 cells and that its expression was not altered after B[a]P treatment. This indicates that TM4 cells are nearly AhR-deficient. In TM4 cells, the CYP1A1 protein and its activity were not present. From these results, it is clear that AhR may be a prerequisite for CYP1A1 expression in TM4 cells. Therefore, TM4 cells can be referred to as CYP1A1-deficient cells. Thus, TM4 Sertoli cells are believed to have a rigid and protective cellular machinery against genotoxic agents. In conclusion, it is suggested that tolerance to B[a]P cytotoxicity is associated with insufficient AhR and CYP1A1 expression in testicular Sertoli cells.

• 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.

• 한국가축번식학회지
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• 제22권4호
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• pp.331-339
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• 1998

본 연구는 Sertoli 세포 column 과 Sertoli 세포돌기 미세구조의 연구를 위해 당질검출을 위한 PA-TCH-SP, -PD 투과전자현미경용 특수염색을 정세관에 적용한 후 각 정세포와 Sertoli 세포를 관찰하고 비교하여 이 분야 연구에 본염색법의 적용 가능성을 검토하였다. PA-TCH-SP, -PD 특수염색을 개의 정세관에 적용하여 투과전자현미경으로 관찰하였던 바, Sertoli 세포 column 과 돌기 부위에 수많은 반응과립이 관찰되었으며 정세관 기저부의 정조세포질에서도 약간의 반응과립이 관찰되었다. 이와 같은 반응과립은 각 정세포의 핵, 정모세포질, 정자세포질 및 잔류체에는 존재하지 않아 반응과립이 존재하는 Sertoli 세포 colunm 과 돌기 및 정자세포질에 침투된 돌기들을 명확히 구분할 수 있었다. 따라서 PA-TCH-SP, -PD 염색법은 투과전자현미경하에서 Sertoli 세포 colunm 과 돌기의 형태학적 및 기능학적 연구에 대단히 유용한 염색법으로 사료된다.

• Clinical and Experimental Reproductive Medicine
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• 제45권2호
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• pp.75-81
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• 2018

Objective: Recapitulation of the spermatogenesis process in vitro is a tool for studying the biology of germ cells, and may lead to promising therapeutic strategies in the future. In this study, we attempted to transdifferentiate Wharton's jelly-derived mesenchymal stem cells (WJ-MSCs) into male germ cells using all-trans retinoic acid and Sertoli cell-conditioned medium. Methods: Human WJ-MSCs were propagated by the explant culture method, and cells at the second passage were induced with differentiation medium containing all-trans retinoic acid for 2 weeks. Putative germ cells were cultured with Sertoli cell-conditioned medium at $36^{\circ}C$ for 3 more weeks. Results: The gene expression profile was consistent with the stage-specific development of germ cells. The expression of Oct4 and Plzf (early germ cell markers) was diminished, while Stra8 (a premeiotic marker), Scp3 (a meiotic marker), and Acr and Prm1 (postmeiotic markers) were upregulated during the induction period. In morphological studies, approximately 5% of the cells were secondary spermatocytes that had completed two stages of acrosome formation (the Golgi phase and the cap phase). A few spermatid-like cells that had undergone the initial stage of tail formation were also noted. Conclusion: Human WJ-MSCs can be transdifferentiated into more advanced stages of germ cells by a simple two-step induction protocol using retinoic acid and Sertoli cell-conditioned medium.

• 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.

• Applied Microscopy
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• 제29권3호
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• pp.353-362
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• 1999

플라스틱 제품의 가소제로 널리 사용되며, 내분비 교란물질로도 알려져 있는 di-(2-ethylhexyl) phthalate (DEHP)를 사춘기 이전 흰쥐에 1주일 동안 구강 투여 (1g/kg, 3g/kg, 5g/kg)한 후 분화중인 정소의 미세구조에 미치는 영향을 조사하였다. DEHP 처리군에서는 대조군에 비하여 세정관 직경이 매우 작았으며, Leydig 세포, Sertoli 세포 및 분화중인 생식세포들의 증식이 억제되었다. 실험군에서 세정관 사이에 존재하여 테스토스테론을 분비하는 Leydig세포는 이질염색질이 증가하고, 세포질내에서 활면소포체의 발달이 미흡하였으며, 리소솜과 지방적이 증가하는 특징을 보였다. 세정관 내에서 Sertoli 세포는 세포질의 크기가 감소하고, 핵막이 불규칙해지거나, 염색질이 응축되어 있는 모습이 나타나며, 세포질내에서 리소솜과 액포들이 증가하였다. 실험군 흰쥐 정소내에서 분화중인 생식세포들의 핵은 이질염색질이 증가하거나, 응축되어 있고, 인의 분리현상이 관찰되기도 하며, 심한 경우 핵내에 공포가 형성되어 있거나, 괴사과정에 있는 생식세포들도 관찰되었다. 이와 같은 정소 미세 구조의 변화는 DEHP 투여량의 농도에 의존성을 보이며, 이 화합물은 Leydig 세포의 테스토스테론 합성 기능을 방해하고, 이어서 Sertoli 세포의 구조와 기능이 손상되어 정자형성과정 중인 생식세포들의 증식과 분화가 억제되는 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.