• Title/Summary/Keyword: Leydig 세포

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Evaluation of Bisphenol a Induced Apoptosis in Sertoli Cell-lines (Bisphenol A에 의한 Sertoli 세포주 내 세포자연사 검정)

  • Kim, Ji-Hyang;Kim, Jin-Kyu;Do, Byoung-Rok;Lee, Chang-Joo;Yoon, Yong-Dal
    • Development and Reproduction
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    • v.9 no.2
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    • pp.115-121
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    • 2005
  • The present report aimed at evaluating the effect of bisphenol A(BPA) and diethylstilbestrol(DES) on Leydig or Sertoli cell-lines. To identify the differences in the susceptibility to BPA upon different cell-types, assay of the cell viability was done on TM3(Leydig cells) and TM4(Sertoli cells) cell-lines. The result indicates that Sertoli cells are more sensitive to low dose of BPA than Leydig cells. Also, the BPA- or DES-treated Sertoli cells showed a reduction of phospholipase D(PLD) activity identically. According to the confirmation of the mRNA expression of fas receptor and fas ligand in the BPA-treated cells, fas/fasL system activated by BPA will deliver the apoptosis signal onto Sertoli TM4 cells. However, Fas/FasL system was not activated in the DES-treated cells unlike the BPA-treated cells.

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Protective effects Acanthopanax senticosus extracts on ethanol-induced apoptosis in TM3 mouse Leydig cells (오가피전탕액(五加皮煎湯液)이 에탄올에 의해 유발되는 TM3 mouse Leydig 세포주의 apoptosis에 미치는 영향)

  • Kim Jong-Chul;Kim Hee-Taek;Kim Ho-Hyun;Kim Ee-Hwa
    • Korean Journal of Acupuncture
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    • v.20 no.1
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    • pp.57-64
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    • 2003
  • 목적: 오가피가 mouse testis에서 유래된 Leydig 세포주에서 에탄올에 의해 유발된 아폽토시스에 미치는 영향을 조사하였다. 방법: TM3 세포주에서의 아폽토시스 변화를 관찰하기 위해서 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, 4,6-diamidino-2-phenylindole (DAPI), DNA fragmentation assay, 및 reverse transcription-polymerase chain reaction (RT-PCR) 방법을 이용하였다. 결과: MTT assay를 이용하여 분석한 결과 농도에 따른 세포독성의 효과가 에탄올 투여부터 관찰되었다. 또한 오가피로 전처치하고 에탄올을 처치하였을 때 세포독성이 크게 감소되었다. DAPI staining에서 오가피 투여군은 에탄올 투여군에 비해서 fragmentation이 억제되었다. RT-PCR의 분석에 의하여 caspase-3 mRNA 발현이 오가피 투여군은 알코올 투여군보다 유의성있게 억제됨을 보여주었다. 결론: TM3 Leydig 세포주에서 에탄올에 의해 유발된 아폽토시스는 전형적인 세포사멸 형태를 나타내었다. 반면에 오가피 투여군은 에탄올에 의해서 유발된 아폽토시스에서 세포보호 효과가 있음이 확인되었다.

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Effects of Bisphenol A on Gene Expression and Apoptosis of Leydig Cells in the Mouse Testis (생쥐 정소에서 비스페놀 에이 (Bisphenol A)가 Leydig Cell의 유전자 발현과 세포자멸사에 주는 영향)

  • Eo, Jin-Won;Lim, Hyun-Jung
    • Clinical and Experimental Reproductive Medicine
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    • v.35 no.3
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    • pp.181-191
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    • 2008
  • Objective: Environmental chemicals alter reproduction, growth, and survival by changing the normal function of the endocrine system. Bisphenol A (BPA), one of the endocrine disruptors, is known to be an estrogen receptor agonist. Therefore, we hypothesized that BPA may affect male reproduction including spermatogenesis in the mouse testis. Methods: We used 7-week-old ICR mice. The first experiment group received BPA in sesame oil (vehicle, 1 mg/kg, 10 mg/kg, and 100 mg/kg) by i.p. injection and mice were sacrificed 24 hr later. The second experiment group received BPA (vehicle, 10 ${\mu}g/kg$, 1 mg/kg, and 100 mg/kg) daily for 14 days by subcutaneous injection. Expression of cell type-specific marker genes in the testis was evaluated by RT-PCR. Histological analysis, immunofluorescence staining, and TUNEL staining were also performed. Results: RT-PCR analyses showed that expression of luteinizing hormone receptor (LHR), a marker gene for the Leydig cell, was notably decreased in the testes of high dose-exposed mice. No obvious difference in the histology of testes was noted among treatment groups. Immunostaining of LHR in the first experiment group did not show noticeable difference in LHR protein expression in Leydig cells. Immunohistochemistry also revealed heightened expression of the immunoreactive Bax in the treatment group, and this was accompanied by positive TUNEL staining in the interstitial area within testis where Leydig cells reside. Conclusions: Our result suggests that BPA affects Leydig cell functions by altering gene expression and by increasing apoptosis in the mouse testis.

Morphological Differentiation of Leydig Cells in Human Fetal Testes (사람 태아 고환에서 간질세포 분화의 형태학적 관찰)

  • Rhee, Kye-Il;Kim, Dae-Joong;Kim, Kyung-Yong;Park, Eon-Sub
    • Applied Microscopy
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    • v.29 no.4
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    • pp.511-522
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    • 1999
  • The Leydig cell found within the interstitium of the testis is important in the spermatogenesis. The differentiation of Leydig cell, even though relatively well known in animals, is not fully elucidated yet in human. In the present study, human fetal testes ($14\sim27$ weeks) obtained from artificially induced abortions legally without gross malformations were studied using light and transmission electron microscopy to make clear the differentiation process of Leydig cells in humans. Leydig cells could be classified as 4 types: fusiform, light, dark, and degenerating cells. The most immature cell was the fusiform cells found frequently at 14 weeks, which seemed to differentiate successively into light and dark cells. Light cells were most frequently found at 17 weeks and dark cells at 24 weeks. Light cells were found to have mitochondria and smooth endoplasmic reticuli (sER) most prominently than any other cell types. The lumen of sER became to be expanded with age. Some electron-dense inclusions were observed in the mitochondrial matrix of the dark cells. Lipid droplets found more in light cells than dark cells were most prominent at 16 weeks and gradually decreased after 20 weeks. Glycogen particles were rich in dark cells. Degenerating cells were most frequently found at 27 weeks. From the results, it is suggested that Leydig cells in human fetal testes undergo similar differentiation process af in animals.

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Effects of Di-(2-ethylhexyl) phthalate (DEHP) on Ultrastructure of Rat Testis (흰쥐 정소의 미세구조에 미치는 Di-(2-ethylhexyl) phthalate (DEHP)의 영향)

  • Kim, Wan-Jong;Kil, Young-Chun;Shin, Kil-Sang
    • Applied Microscopy
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    • v.29 no.3
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    • pp.353-362
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    • 1999
  • Di-(2-ethylhexyl) phthalate (DEHP) is a plasticize. known as one of endocrine disruptors. The present study was carried out to investigate the ultrastructural changes of prepubertal rat testis after oral administration of DEHP in dosages of 1 g/kg, 3 g/kg or 5g/kg in 0.5 ml of corn oil daily for a week. This study revealed the DEHP inhibited the development of seminiferous tubules and induced structural changes on various cell types of the rat testis. Leydig cells, Sertoli cells and the developing germ cells seemed to be impaired their differentiations in terms of the structural changes of cell organelles. The increase of heterochromatin in amount were common features in all 3 cell types. In addition, the Leydig cells were characterized by the increases in number and size of lysosomes and the scantiness of smooth endoplasmic reticulum. The Sertoli cells became irregular in nuclear envelope and the cytoplasm decreased, but the number of lysosomes and vacuoles seemed to be increased. There were some indications of necrosis of the germ cells, such as vacuolized nucleus and segregated nucleolus. These detrimental effects of DEHP on the rat testis were dose dependent and suppressed spermatogenesis decreasing developing germ cells in number and appearances. The effect of DEHP on ultrastructure of rat testis, as its known physiological functions, seems come from the decreased level of testosterone by Leydig cells, followed by the abnomalities of Sertoli cells and the germ cells.

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옻나무 유래 Flavonoid처리가 흰쥐 Leydig 세포의 체외배양에서 Testosterone분비에 미치는 영향

  • 성환후;최선호;민관식;장유민;나천수;정일정
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.80-80
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    • 2001
  • 옻나무 유래 Flavonoid(F)는 항암 효과, 숙취해소 등의 효능이 있으며, 신장의 기능을 촉진하고 생식에도 영향을 준다고 알려져 있다. 그 주성분으로는 fustin, fisetin, sulfuretin, butein으로 추정되고 있다. 본 연구팀의 예비실험에서 성숙 흰쥐에게 F를 30일간 5mg씩 구강투여 하여 정소 중량 및 혈액 내 Testosterone 농도를 분석한 결과, 정소 중량이 증가되고 혈중 Testosterone의 농도도 대조구에 비해 유의적으로 높은 결과를 얻었다. 이에 본 연구의 목적은 옻나무 유래 F가 성숙 수컷 횐쥐의 생식 기능에 직접 미치는 영향을 알아보고자, 흰쥐 Leydig 세포를 분리, 회수하여 체외배양에서 Testosterone농도를 조사하였다. SD계 흰쥐(약 12주령, 체중 250g 전후)의 정소를 적출 한 후, Leydig 세포를 회수하기 위해 0.25% collagenase용액에 넣어 34$^{\circ}C$에서 15분간 진탕 수조에서 배양하였다. 배양된 정소조직은 D-MEM 배양액(10% FCS와 antibiotics 첨가)으로 2~3회 세척한 후 세포 수를 1$\times$$10^{6}$ cells/$m\ell$/well로 분주하여, F(20, 40, 80, 160ng), IGF-I(50, 100ng) 와 LH(10, 100ng) 용량별 각각 첨가하여 배양하였다. 각 처리 후, 배양시간(3, 6, 12, 24, 36시간)에 따라 배양액을 회수하여 COAT-A-COUNT(DPC, USA) kit를 이용하여 RIA방법에 의해 Testosterone을 분석한 결과는 다음과 같다. 대조구 Leydig 세포를 36시간까지 체외 배양하여 Testosterone의 농도를 조사한 결과, 24시간 배양구가 가장 높은 농도를 나타내었다. F를 단독 첨가하여 12시간 배양한 실험에서 F 80ng 첨가구에서 유의적으로 높은 농도를 보였다. LH 10ng 및 100ng 첨가구의 시간별 변화로는, LH 10ng 첨가구에서는 6~12시간 이후, LH 100ng 첨가구에서는 3~6시간 사이에서 유의적인 증가를 보였다. 또한, LH 10ng 첨가 실험에서는 LH 10ng+F 40ng에서 12시간 배양 시, 유의적으로 높은 Testosterone 분비를 확인하였으며, LH 100ng을 첨가하여 3시간 배양 시는 각 처리구 마다 높은 유의적 차이를 보였다. 한편, GH에 관여하는 IGF-I 첨가효과를 비교 검토한 결과, LH 100ng+IGF-I 100ng, 6시간 배양구에서 유의적으로 높게 나타내었다. 이상의 결과로, F 단독 처리 시의 적정량은 약 80ng이며, 흰쥐 정소 Leydig 세포의 Testosterone분비를 촉진하는 작용을 하는 것으로 사료되며, 특히 LH+F구에서 Testosterone분비를 더욱 향상시킴으로써 정소 Leydig 세포의 Androgen 생성을 촉진시키는 역할이 있는 것으로 보여진다 따라서 옻나무 유래 F는 포유동물의 생식기능에 중요하게 작용하는 것으로 사료된다.

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Establishment of Incubational Conditions for Rat Testicular Cells (랫드 고환세포의 배양조건 설정에 관한 연구)

  • 김판기;박귀례;한순영;신재호;이유미;김준규;권석철;이용욱;장성재
    • Journal of Environmental Health Sciences
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    • v.21 no.1
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    • pp.68-73
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    • 1995
  • This study of culturing testicular cell types in vitro has potential to be an invaluable tool for assessing the mechanisms of testicular toxicity, especially those of intragonadal interaction and spermatogenesis. Combined with the Sertoli/germ cell cultures, Leydig cells provide comprehensive and detailed information on the action of testicular toxicants at the level of the testis. Sertoli/germ cell were isolated and incubated well in vitro from 20~30 g rats and Leydig cells from 250~300 g rats. The Sertoli cells isolated from the testis of the SD rats grew into monolayer on about the 2nd~3rd day of culture, an appreciable cell increment being observed between the 4th~5th day. The Leydig cells isolated from the testis of the SD rats grew into a monolayer on about the 3rd-4th day of culture, an appreciable cell increment being observed between the 5th-7th day. These results suggest that Sertoli and Leydig cells can be cultured as a male fertility evaluation method alternative to the in vivo/conventional fertility test method and further study for the physio-chemical determination is needed.

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Tributyltin Increases Adipogenesis and Apoptosis of Rat Testicular Interstitial Cells (Tributyltin에 의한 흰쥐 정소 내 간질세포의 지방세포 유도와 세포자연사 증가)

  • Song, Yeon-Hwa;Jung, Ji-Eun;Lee, Hyun-A;Hong, Ji-Hee;Yang, Hyun-Won
    • Development and Reproduction
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    • v.14 no.4
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    • pp.297-306
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    • 2010
  • Tributyltin (TBT), an endocrine disrupting chemical, has been reported to decrease testicular function by causing apoptosis in the testis, but this mechanism is not fully understood. Thus, in this study we examined whether TBT induces adipogenesis of the Leydig cells to find out the correlation between adipogenesis and apoptosis in the testis. Three week old SD male rats were orally administrated with sesame oil, 1 mg/kg of TBT, or 10 mg/kg of TBT daily for 1 week and weighed after administration. The testes obtained on day 8 were weighed and stained with BODIPY and TUNEL kit. Using total RNA extracted from the isolated Leydig cells, adipogenesis and apoptosis-related genes were analyzed by real-time PCR. The testicular weights of the rats treated with 10 mg/kg TBT were significantly decreased compared to those in the control rats treated with sesame oil. As a result of BODIPY staining, the number of Leydig cells stained with BODIPY was increased in the rats treated with 10 mg/kg TBT compared with the control rats. Similar to BODIPY staining results, the TUNEL assay showed that the apoptosis of Leydig cells was increased in TBT treated rats. The results of the gene expression analysis in the Leydig cells showed that the expression of adipogenesis-related genes (PPAR${\gamma}$, aP2, Perilipin, CD36) and apoptosis-related genes (TNFRSF1A, TNFSF10) was increased after TBT administration. The present study demonstrates that TBT induces the expression of adipogenesis-related and apoptosis-related genes in the Leydig cells leading to adipogenesis and apoptosis in the testes. These results suggest that the dysfunction of Leydig cells by TBT exposure may cause a loss in testicular function.

The Ultrastructure of Leydig Cells in the Testis of Korean Native Chickens (한국 재래 닭 고환 간질세포의 미세구조에 관한 연구)

  • Jang, B.G.;Tae, H.J.;Choi, C.H.;Park, Y.J.;Yang, H.H.;Kim, N.S.;Park, S.Y.;Kang, H.S.;Park, B.Y.;Lee, Y.H.;Ahn, D.C.;Kim, I.S.
    • Korean Journal of Poultry Science
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    • v.33 no.3
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    • pp.181-188
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    • 2006
  • Changes in the fine structure of testicular Leydig cell from hatching to adulthood were studied in Korean native chickens of 1, 2, 4, 6, 8, 10, 12, 14, 16, 18, 21, 24, 28, 32, 44, 52 and 64 weeks (n=13 chickens per group) of age. The objective of this study were to elucidate Leydig cell ultrastructure during testicular development. Testes of chickens were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in Epon-araldite. Using $1{\mu}m$ sections stained with methylene blue-azure II, qualitative and quantitative(stereological) morphological studies were performed. The ultrastructural changes of the Leydig cell were investigated by ultrathin section with the transmission electron microscope. The stages of the Leydig cell development described focus on mitochondria, endoplasmic reticulum, and lipid droplets which are involved in androgens as fullows. 1) Approaching puberty. The closely packed Leydig cells and sparse intercellular space. The nucleus occupied a large portion of the Leydig cell volume. The population of Leydig cells contained two types of cells that differed in the appearance of their nuclei which were either highly electron-opaque or relatively electron-lucid. The cytoplasm was characterized by large amounts of lipid droplets, relatively few spherical mitochondria, and sparse smooth endoplasmic reticulum. 2) Puberty to adult. The Leydig cells which display features compatible with significant androgen synthesis: large volume of cytoplasm containing extended smooth endoplasmic reticulum, abundant mitochondria, and reduction of lipid droplets.

Effect of Di-(2-ethylhexyl)phthalate(DBHP) on Spermatogenesis in Rat Testes (흰쥐 정자형성과정에 미치는 Di-(2-ethylhexyl)phthalate의 영향)

  • 김완종;길영천;이종화;신길상
    • Korean Journal of Environmental Biology
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    • v.17 no.3
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    • pp.285-292
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    • 1999
  • Di-(2-ethylhexyl) phthalate (DEHP) is a plasticizer known as one of endocrine disruptors. The present study was carried out to investigate the alterations of function and ultrastructure in rat testes after oral intubation of DEHP in dosages of 1g/kg/day, 2 g/kg/day or 3 g/kg/day in 0.5 ml of corn oil for 15 days. DEHP reduced the growth of body and testes, inhibited apermatogenesis and induced structural changes on various cell types of the rat testis. Leydig cells, Sertoli cells and the developing germ cells seemed to be impaired their differentiations in terms of the structural changes of cell organelles. The increase of heterochromatin in amount were common features in all 3 cell types. In addition, the Leydig cells were characterized by the swelling of smooth endoplasmic reticulum and perinuclear space, the increases in number and size of Iysosomes. The Sertoli cells became irregular in nuclear envelope and the number of Iysosomes and vacuoles seemed to be increased. There were some indications of necrosis of the germ cells, such as vacuolized nucleus and segregated nucleolus. And also, DEHP lowered the level of testosterone in experimental rat serum. DEHP suppressed apermatogenesis decreasing developing germ cells and these effects of DEHP on the rat testis were dose dependent. The detrimental effect of DEHP on apermatogenesis and ultrastructure of rat testes seems to be derived from the decreased level of testosterone by Leydig cells, followed by the abnomalities of Sertoli cells and the germ cells.

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