• Korean Journal of Veterinary Research
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• v.42 no.3
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• pp.299-308
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• 2002

The purpose of this morphometric study was to obtain detailed quantitative information on all cell types in the testis interstitium of Korean ring-necked pheasants combined with data on changes in the steroidogenic function of the testis during the breeding and nonbreeding seasons. Animals collected during the breeding season, testis weights, sperm production, serum testosterone levels, leuteinizing hormone-stimulated testosterone secretion, and the length of the seminiferous tubules were significantly (p < 0.05) increased as compared to the nonbreeding season. Seminiferous tubules occupied 93.25% of testis volume in the breeding season. Leydig cells constituted 0.82% of the testicular volume. The mean volume of an Leydig cell was $1039{\mu}m^3$, and each testis contained about 24.53 million Leydig cells. Testes of the pheasants during the nonbreeding season displayed a 98% reduction in testis volume that was associated with a decrease in the absolute volume of seminiferous tubules (98% reduction), tubular lumen(100%), interstitium(90%), blood vessels(84%), lymphatic spaces(97%), Leydig cells(79%), mesenchymal cells(51%), and myoid cells(61%). The number of Leydig cells, mesenchymal cells, myoid cells per testis in the breeding season was higher (p < 0.05) than in the nonbreeding season. Although the average volume of a Leydig cell was 74% lower in the nonbreeding season, the average volume of a myoid and mesenchymal cell remained unchanged. These results demonstrate that there are a striking differences in the testicular structure of the Korean ring-necked pheasant in the breeding and nonbreeding seasons. Every structural parameter of the Leydig cell was pasitively correlated with both serum and LH-stimulated secretion concentrations of testosterone. Correlation of changes in hormonal status with morphometric alterations of all Leydig cell suggests that the Korean-ring necked pheasant may be used as a model to study structure-function relations in the avian testis.

• The Korea Journal of Herbology
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• v.26 no.1
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• pp.133-138
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• 2011

Objectives : The purpose of this study was to estimate the antioxidant activity of Sesami Semen Nigrum extract (SSN) on mouse Leydig cells, TM3. Methods : Cell viability assays were performed. The protective effects of SSN against hydrogen peroxide-induced oxidative stress in Leydig cells were examined by measuring cell viability. Lipid peroxidation levels and antioxidant enzyme concentrations such as SOD and catalase were measured. Results : Cell viability of Leydig cells increased with SSN concentration. Cell viability of Leydig cells was 136.66% when SSN concentration was $50{\mu}g/ml$. Cell viability of the hydrogen peroxide group was statistically decreased (p<0.01) compared with the control group. Antioxidant effect of SSN was measured and the protective effect of SSN concentration were 5, 10, $50{\mu}g/ml$. LPO were decreased significantly at 5, $50{\mu}g/ml$ of SSN concentrations. SOD activity was increased at 1, 10, $50{\mu}g/ml$ of SSN concentrations. Catalase activity was significantly increased at 123.7, 133.3 and 131.9 units/mg protein when SSN concentrations were 5, 10 and $50{\mu}g/ml$, respectively. Conclusions : In conclusion, Sesami Semen Nigrum extract has antioxidant activities in Leydig cells against oxidative stress.

• Herbal Formula Science
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• v.23 no.1
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• pp.111-119
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• 2015

Objectives : The purpose of this study was to investigate the antioxidant activity of water extract of Woogyuyeum (WGY) in Leydig cells. Methods : We investigated the cytoprotective effect of WGY in cultured mouse Leydig cells by MTT assay. Leydig cells treated with WGY were incubated in the presence or absence of 50 μM hydrogen peroxide at 37℃ for 24 h. The protective effects of WGY against hydrogen peroxide-induced oxidative stress, lipid peroxide (LPO), superoxide dismutase (SOD), and catalase activity assays were performed in Leydig cells. Results : As a result, WGY showed no significant cytotoxicity in Leygdig cells. WGY showed cell viability as 103.65% in 5 μg/ml concentrations. The cytotoxicity induced by hydrogen peroxide in Leygdig cells, the antioxidant effects of WGY was increased in 1, 5, 50, 100 ug/ml concentraions. 100 μg/ml concentration of WGY showed maximum antioxidant effects. Treatment of cells with 100 μg/ml WGY significantly reduced the MDA concentration to 0.23 nmoles/mg protein. SOD activity was increased at 1, 100 μg/ml concentration of WGY and catalase activity was significantly increased at 50, 100 μg/ml concentrations of WGY, respectively. Conclusions : In conclusion, WGY has antioxidant activities against hydrogen peroxide-induced oxidative stress in Leydig cells.

• Journal of Animal Reproduction and Biotechnology
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• v.36 no.3
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• pp.145-153
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• 2021

The role of transient receptor potential vanilloid receptor-1 (TRPV1) has been primarily investigated in pain sensory neurons. Relatively, little research has been performed in testicular cells. TRPV1 is abundantly expressed in Leydig cells of young adult mice. This study was conducted to determine the role of the TRPV1 channel in Leydig cells. TRPV1 modulators and testosterone were treated to the mouse Leydig cell line TM3 cells for 24 h. Capsaicin, a TRPV1 activator, dose-dependently induced cell death, whereas capsazepine, a TRPV1 inhibitor, inhibited capsaicin-induced cell death. Testosterone treatment reduced capsaicin-induced cell death. High concentrations of testosterone decreased TRPV1 mRNA and protein expression levels. However, TRPV1 modulators did not affect testosterone production. These results showed that capsaicin induced cell death of Leydig cells and that testosterone reduced capsaicin-induced cell death. Our findings suggest that testosterone may regulate the survival of Leydig cells in young adult mice by decreasing the expression level of TRPV1.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.5
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• pp.618-625
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• 2009

The objective of this work was to study the direct effects of daidzein on steroidogenesis in cultured mouse Leydig cells. Adult mouse Leydig cells were purified by Percoll gradient centrifugation, and the cell purity was determined using a $3{\beta}$-hydroxysteroid dehydrogenase ($3{\beta}$-HSD) staining method. The purified Leydig cells were exposed to different concentrations ($10^{-7}$ M to $10^{-4}$ M) of daidzein for 24 h under basal and human chorionic gonadotropin (hCG)-stimulated conditions. The cell viability and testosterone production were determined, and the related mechanisms of daidzein action were also evaluated using the estrogen receptor antagonist ICI 182,780 and measuring the mRNA levels of steroidogenic acute regulatory protein (StAR), cholesterol side-chain cleavage enzyme (P450scc), and $3{\beta}$-HSD-1 involved in testosterone biosynthesis. The results revealed that daidzein did not influence cell viability. Daidzein increased both basal and hCG-stimulated testosterone production in a dose-dependent manner, and this effect was statistically significant at concentrations of $10^{-5}$ M and $10^{-4}$ M daidzein (p<0.05). ICI 182,780 had no influence on daidzein action. RTPCR results revealed that $10^{-5}$ M and $10^{-4}$ M daidzein did not exert any obvious influence on the mRNA level of P450scc in Leydig cells. However, in the presence of hCG, these concentrations of daidzein significantly increased the StAR and $3{\beta}$-HSD-1 mRNA levels (p<0.05), but in the absence of hCG, only $10^{-5}$ M and $10^{-4}$ M daidzein up-regulated the StAR and $3{\beta}$-HSD-1 mRNA expression (p<0.05), respectively. These results suggest that daidzein has direct effect on Leydig cells. Daidzein-induced increase of testosterone production is probably not mediated by the estrogen receptor but correlates with the increased mRNA levels of StAR and $3{\beta}$-HSD-1.

• 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.

• Environmental Analysis Health and Toxicology
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• v.22 no.1 s.56
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• pp.57-64
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• 2007

Nanomaterials have been used to create unique devices at the nanoscale level. However, the toxicities of nanomaterials have not been fully tested and the risk of nanomaterials has been raised as an emerging issue in these days. In this study, the cytotoxicity of silver nanoparticles was tested using cultured mouse leydig cells. As results, silver nanoparticles showed cytotoxicity with the generation of reactive oxygen species (ROS). With the increased level of ROS, intracellular glutathione level was decreased. DNA fragmentation and caspase-3 activation suggested the apoptotic mechanism of cell death in leydig cells treated with silver nanoparticles.

• Development and Reproduction
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• v.18 no.4
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• pp.187-195
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• 2014

Polycyclic aromatic hydrocarbons (PAHs), which are ubiquitous in the air, are present as volatile and particulate pollutants that result from incomplete combustion. Most PAHs have toxic, mutagenic, and/or carcinogenic properties. Among PAHs, benzo[a]pyrene (B[a]P) and dimethylbenz[a]anthracene (DMBA) are suspected endocrine disruptors. The testis is an important target for PAHs, yet effects on steroidogenesis in Leydig cells are yet to be ascertained. Particularly, disruption of testosterone production by these chemicals can result in serious defects in male reproduction. Exposure to B[a]P reduced serum and intratesticular fluid testosterone levels in rats. Of note, the testosterone level reductions were accompanied by decreased steroidogenic acute regulatory protein (StAR) and $3{\beta}$-hydroxysteroid dehydrogenase isomerase ($3{\beta}$-HSD) expression in Leydig cells. B[a]P exposure can decrease epididymal sperm quality, possibly by disturbing the testosterone level. StAR may be a key steroidogenic protein that is targeted by B[a]P or other PAHs.

• 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.

• The Korea Journal of Herbology
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• v.36 no.6
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• pp.17-25
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• 2021

Objectives : Baicalein (3,3', 4', 5, 6-pentahydroxyflavone), a type of flavonoid, is a well-known antioxidant and anti-inflammatory ingredient found in Scutellaria baicalensis root. The aim of this study is to investigate the effect of baicalein on nitric oxide (NO) production in TM3 mouse Leydig cells stimulated by indomethacin (IN). Methods : TM3 cells were treated with IN (0.5 μM) and baicalein at concentrations of 12.5, 25, 50, and 100 μM for 24 hr, 40 hr, 42 hr, 44 hr, and 64 hr. After treatments, cell viabilities were measured with the modified MTT assay. The production of nitric oxide in cells was measured by Griess reagent assay. Results : Baicalein showed no cytotoxicity on IN-stimulated TM3. NO production in IN-stimulated TM3 treated for 24 hr with baicalein at concentrations of 12.5, 25, 50, and 100 μM was 95.8%, 94.86%, 89.97%, and 81.52% of the control group treated with IN only, respectively; NO production for 40 hr was 97.34%, 97.34%, 95.15%, and 87.42%, respectively; NO production for 42 hr was 89.12%, 90.14%, 89.74%, and 90.26%, respectively; NO production for 44 hr was 83.83%, 84.94%, 85.65%, and 86.85%, respectively; NO production for 64 hr was 94.12%, 95.38%, 94.21%, and 94.12%, respectively. Specifically, baicalein at concentrations of 12.5, 25, and 50 have been shown to most efficiently inhibit NO productions in 48 hr of treatment. Conclusions : Baicalein might have anti-toxicant effect on Leydig cells related with its inhibition of NO production in Leydig cells stimulated with IN.