Uptake in Vivo of [3h]testosterone by the Interstitial Compartment in Testes of Normal Adult Mice

Beardsley, J. A.; Hilton, Frederick K.

doi:10.1530/jrf.0.0420361

Cited by 6 publications

(6 citation statements)

References 10 publications

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“…It was suggested that androgen's action upon these cells may be required for the maintenance of their steroid-synthesizing enzymes. 21 In our unpublished preliminary survey on endogenous testosterone secreted by TM3 cells, we found that testosterone is increased in secretion between 50 to 500 nmol l 21 of exogenous supplementation of testosterone. A time-dependant increase in secretion under 100-nmol l 21 supplementation of testosterone was found in cells from 6 to 48 h (60-96 ng dl 21 ).…”

Section: Discussionmentioning

confidence: 79%

“…The TM3 cell line was cultured in medium with a 1 : 1 mixture of phenol red-free Ham's F12 medium and phenol red-free Dulbecco's modified Eagle's medium with 4.5 g l 21 glucose, 1.2 g l 21 sodium bicarbonate, 15 mmol l 21 HEPES, 100 nmol l 21 of ICI 182780 and 5% fetal bovine serum (Gibco, Invitrogen, Grand Island, NY, USA). For testosterone supplementation studies, testosterone (Organon, Oss, The Netherlands) was administered at the doses of 10, 50, 100, 500, 1000 and 2000 nmol l 21 . For time-related studies, testosterone was administered at a dose of 100 nmol l 21 for 6, 8, 12, 24, 36 and 48 h.…”

Section: Cell Culture and Preparationmentioning

confidence: 99%

“…After an additional hour of incubation, 150 ml of dimethyl sulfoxide was added to the wells to extract the formazan. The formazan absorbance was measured at 565 nmol l 21 , and values represent the optical density per mg of protein.…”

Section: Cell Viabilitymentioning

confidence: 99%

“…After cells were 90% confluent, they were washed three times with Hank's balanced salt solution. Following the addition of 10 mmol l 21 H 2 DCFDA (final concentration), the appearance of DCF fluorescence was typically followed for 1 h, using excitation at 486 nmol l 21 and emission at 530 nmol l 21 . The fluorescence profiles were quantified using the LYSYS II software program (Becton Dickenson, Franklin Lakes, NJ, USA).…”

Section: Cell Viabilitymentioning

confidence: 99%

“…After mixing, 0.2 ml of 42-mmol l 21 thiobarbituric acid solution was added to a final concentration of 7 mmol l 21 and then placed in a 95 uC dry bath for 1 h. Samples were then cooled and neutralized with 1 mol l 21 NaOH in methanol before the high-performance liquid chromatographic analysis. An aliquot of 20 ml of supernatant obtained above was injected into a narrow-pore C 18 column (4.63250 mmol l 21 , with a particle size of 5 mmol l 21 ) using a Jasco PU-980 pump (Jasco, Tokyo, Japan) with a solvent system composed of methanol and 50 mmol l 21 phosphate buffer (pH 6.8) (4 : 6, v/v) at a flow rate of 1 ml min 21 . The eluent was monitored with a Jasco fluorescence detector (with excitation at 525 nmol l 21 and emission at 550 nmol l 21 ).…”

Section: Determination Of Lipid Peroxidementioning

confidence: 99%

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Low-dose testosterone treatment decreases oxidative damage in TM3 Leydig cells

Hwang¹,

Liao²,

Lin³

et al. 2011

Asian J Androl

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Testosterone replacement therapy has benefits for aging men and those with hypogonadism. However, the effects of exogenous testosterone on Leydig cells are still unclear and need to be clarified. In this report, we demonstrate that testosterone supplementation can reduce oxidative damage in Leydig cells. The TM3 Leydig cell line was used as an in vitro cell model in this study. Cytoprotective effects were identified with 100-nmol l 21 testosterone treatment, but cytotoxic effects were found with o500-nmol l 21 testosterone supplementation. Significantly reduced reactive oxygen species (ROS) generation, lipid peroxide contents and hypoxia induction factor (HIF)-1a stabilization and activation were found with 100-nmol l 21 testosterone treatment. There was a 1.72-fold increase in ROS generation in the 500-nmol l 21 compared to the 100-nmol l 21 testosterone treatment. A 1.58-fold increase in steroidogenic acute regulatory protein (StAR) expression was found in 50-nmol l 21 testosterone-treated cells (P,0.01). Chemically induced hypoxia was attenuated by testosterone supplementation. Leydig cells treated with low-dose testosterone supplementation showed cytoprotection by decreasing ROS and lipid peroxides, increasing StAR expression and relieving hypoxia stress as demonstrated by HIF-1a stabilization. Increased oxidative damage was found with o500-nmol l 21 testosterone manipulation. The mechanism governing the differential dose effects of testosterone on Leydig cells needs further investigation in order to shed light on testosterone replacement therapy. INTRODUCTIONTestosterone plays an important role in normal growth and development of male sex organs. It is also known to be a key player in glucose homeostasis and lipid metabolism. 1 More recently, gonadotrophinreleasing hormones I and II were demonstrated to stimulate testosterone production in murine Leydig cells. 2 In the testes, Leydig cells synthesize and secrete testosterone in response to luteinizing hormone, which is counter-regulated by feedback influences of testosterone and its metabolites. Testosterone levels notably decline with aging; 3,4 however, a recent report showed no correlation between androgen receptor (AR) polymorphism and the serum concentrations of total and free testosterone in elderly men. 5 The mechanism by which aged Leydig cells lose their steroidogenic function remains unclear. It was proposed that decreases in testosterone levels with aging may reflect reactive oxygen species (ROS) elevation and further disruption of the ability of Leydig cells to produce testosterone. 6,7 Mitochondrial respiration, which typically consumes about 90% of the oxygen utilized by cells, is considered to be the major source of cellular ROS. 8 ROS cause tissue damage by a variety of mechanisms including DNA damage, lipid peroxidation, protein oxidation and carbonylation, depletion of cellular thiols and activation of proinflammatory cytokine release. Mitochondrial-derived ROS were implicated in a number of disease and disorders, including neurodegenerat...

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Section: Discussionmentioning

confidence: 79%

Section: Cell Culture and Preparationmentioning

confidence: 99%

Section: Cell Viabilitymentioning

confidence: 99%

Section: Cell Viabilitymentioning

confidence: 99%

Section: Determination Of Lipid Peroxidementioning

confidence: 99%

See 3 more Smart Citations