Steroidogenic enzyme activity in the rat testis following Leydig cell destruction by ethylene-1,2-dimethanesulphonate and during subsequent Leydig cell regeneration

1991 ◽  
Vol 131 (3) ◽  
pp. 451-457 ◽  
Author(s):  
P. J. O'Shaughnessy ◽  
L. Murphy
Keyword(s):  
Enzyme Activity ◽  
Leydig Cell ◽  
Leydig Cells ◽  
Seminiferous Tubules ◽  
Cell Regeneration ◽  
Significant Activity ◽  
Steroidogenic Enzyme ◽  
Control Activity ◽  
Rat Testis ◽  
Cell Destruction

ABSTRACT Ethylene-1,2-dimethanesulphonate (EDS) rapidly destroys Leydig cells in the rat testis, although repopulation occurs within 5–7 weeks. In this study we have examined the activity of testicular steroidogenic enzymes after Leydig cell destruction and during regeneration. This was designed to measure the contribution of cells, other than Leydig cells, to steroidogenic activity in the testis, and to determine whether changes in steroidogenic enzyme activity during Leydig cell regeneration after EDS parallel those which occur during normal Leydig cell development. The enzymes studied are those responsible for androgen synthesis and metabolism in the testis. Adult male Wistar rats (300–350 g) were injected with EDS (100 mg/kg, i.p.) and testicular steroidogenic enzyme activity was measured on days 0, 3, 7, 14, 21 and 35. On day 3, when no Leydig cells remain in the testis, cholesterol side-chain cleavage (CSCC) activity, per testis, declined to undetectable levels, while 3βhydroxysteroid dehydrogenase (3βHSD) and 17α-hydroxylase retained only 0·04 and 0·15% of control activity respectively. In contrast, 17-ketosteroid reductase (17-KSR) and 5α-reductase retained 33 and 10% of control activity respectively. On day 7 there was a further loss of 17-KSR activity (to 20% of control) but no change in other enzymes. The 17-KSR activity remaining on day 7 after EDS was contained almost exclusively in the seminiferous tubules, while the low 3β-HSD activity remaining was confined largely to the interstitial tissue. Other enzymes showed a more even distribution between the two compartments. On day 14 after EDS there was a tenfold increase in 3β-HSD activity (compared with day 7), with no change in CSCC, 17α-hydroxylase or 5α-reductase and a further loss of 17-KSR (to 11% of control). Between days 14 and 21 there were marked increases in the activities of CSCC, 3β-HSD, and 17α-hydroxylase, while 17-KSR showed no change in activity from day 14. Activity of 5α-reductase increased between days 14 and 21 to levels greater than those seen in control animals. By day 35 the activities of all enzymes had returned to control levels. The results show that, in the adult rat testis, the activities of CSCC, 3β-HSD and 17α-hydroxylase are confined, almost entirely, to the Leydig cells, and only 17-KSR shows significant activity in another cell type. During regeneration of Leydig cells after EDS the pattern of changes in enzyme activity is very similar to that seen in the normal development of the adult population of Leydig cells. Journal of Endocrinology (1991) 131, 451–457

scholarly journals Influence of long-term dietary administration of procymidone, a fungicide with anti-androgenic effects, or the phytoestrogen genistein to rats on the pituitary–gonadal axis and Leydig cell steroidogenesis

2005 ◽  
Vol 187 (1) ◽  
pp. 117-124 ◽  
Author(s):  
K Svechnikov ◽  
V Supornsilchai ◽  
M-L Strand ◽  
A Wahlgren ◽  
D Seidlova-Wuttke ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Serum Level ◽  
Leydig Cell ◽  
Leydig Cells ◽  
Ex Vivo ◽  
Elevated Serum ◽  
Seminiferous Tubules ◽  
Dibutyryl Camp

Procymidone is a fungicide with anti-androgenic properties, widely used to protect fruits from fungal infection. Thereby it contaminates fruit products prepared for human consumption. Genistein-containing soy products are increasingly used as food additives with health-promoting properties. Therefore we examined the effects of long-term dietary administration (3 months) of the anti-androgen procymidone (26.4 mg/animal per day) or the phytoestrogen genistein (21.1 mg/animal per day) to rats on the pituitary-gonadal axis in vivo, as well as on Leydig cell steroidogenesis and on spermatogenesis ex vivo. The procymidone-containing diet elevated serum levels of LH and testosterone and, furthermore, Leydig cells isolated from procymidone-treated animals displayed an enhanced capacity for producing testosterone in response to stimulation by hCG or dibutyryl cAMP, as well as elevated expression of steroidogenic acute regulatory protein (StAR), cytochrome P450 side-chain cleavage (P450 scc) and cytochrome P450 17α (P450c17). In contrast, the rate of DNA synthesis during stages VIII and IX of spermatogenesis in segments of seminiferous tubules isolated from genistein-treated rats was decreased without accompanying changes in the serum level of either LH or testosterone. Nonetheless, genistein did suppress the ex vivo steroidogenic response of Leydig cells to hCG or dibutyryl cAMP by down-regulating their expression of P450 scc. Considered together, our present findings demonstrate that long-term dietary administration of procymidone or genistein to rats exerts different effects on the pituitary–gonadal axis in vivo and on Leydig cell steroidogenesis ex vivo. Possibly as a result of disruption of hormonal feedback control due to its anti-androgenic action, procymidone activates this endocrine axis, thereby causing hyper-gonadotropic activation of testicular steroidogenesis. In contrast, genistein influences spermatogenesis and significantly inhibits Leydig cell steroidogenesis ex vivo without altering the serum level of either LH or testosterone.

scholarly journals Morphometric analysis of Leydig cells in the normal rat testis.

1980 ◽  
Vol 84 (2) ◽  
pp. 340-354 ◽  
Author(s):  
H Mori ◽  
A K Christensen
Keyword(s):  
Plasma Membrane ◽  
Surface Area ◽  
Morphometric Analysis ◽  
Leydig Cell ◽  
Leydig Cells ◽  
Steroidogenic Enzymes ◽  
Inner Membrane ◽  
Rat Testis ◽  
Mitochondrial Inner Membrane ◽  
Testis Tissue

Leydig cells are thought to be the source of most, if not all, the testosterone produced by the testis. The goal of this study was to obtain quantitative information about rat Leydig cells and their organelles that might be correlated with pertinent physiological and biochemical data available either now or in the future. Morphometric analysis of Leydig cells in mature normal rats was carried out on tissue fixed by perfusion with buffered glutaraldehyde, and embedded in glycol methacrylate for light microscopy and in Epon for electron microscopy. In a whole testis, 82.4% of the volume was occupied by seminiferous tubules, 15.7% by the interstitial tissue, and 1.9% by the capsule. Leydig cells constituted 2.7% of testicular volume. Each cubic centimeter (contained approximatelyy 1 g) of rat testis contained about 22 million Leydig cells. An average Leydig cell had a volume of 1,210 micron3 and its plasma membrane had a surface area of 1,520 micron2. The smooth endoplasmic reticulum (SER), the most prominent organelle in Leydig cells and a major site of steroidogenic enzymes, had a surface area of approximately 10,500 micron2/cell, which is 6.9 times that of the plasma membrane and is 60% of the total membrane area of the cell. The total surface area of Leydig SER per cubic centimeter of testis tissue is approximately 2,300 cm2 or 0.23 m2. There were 3.0 mg of Leydig mitochondria in 1 g of testis tissue. The average Leydig cell contained approximately 622 mitochondria, measuring on the average 0.35 micron in diameter and 2.40 micron in length. The mitochondrial inner membrane (including cristae), another important site of steroidogenic enzymes, had a surface area of 2,920 micron2/cell, which is 1.9 times that of the plasma membrane. There were 644 cm2 of inner mitochondrial membrane/cm3 of testis tissue. These morphometric results can be correlated with published data on the rate of testosterone secretion to show that an average Leydig cell secretes approximately 0.44 pg of testosterone/d or 10,600 molecules of testosterone/s. The rate of testosterone production by each square centimeter of SER is 4.2 ng/d or 101 million molecules/s: the corresponding rate for each square centimeter of mitochondrial inner membrane is 15 ng testosterone/d or 362 million molecules/s.

scholarly journals Effects of Estradiol and Methoxychlor on Leydig Cell Regeneration in the Adult Rat Testis

2014 ◽  
Vol 15 (5) ◽  
pp. 7812-7826 ◽  
Author(s):  
Bingbing Chen ◽  
Dongxin Chen ◽  
Zheli Jiang ◽  
Jingyang Li ◽  
Shiwen Liu ◽  
...  
Keyword(s):  
Leydig Cell ◽  
Cell Regeneration ◽  
Rat Testis ◽  
Adult Rat

scholarly journals Specific protein synthesis in isolated rat testis leydig cells. Influence of luteinizing hormone and cycloheximide

1977 ◽  
Vol 162 (2) ◽  
pp. 341-346 ◽  
Author(s):  
F H A Janszen ◽  
B A Cooke ◽  
H J van der Molen
Keyword(s):  
Protein Synthesis ◽  
Luteinizing Hormone ◽  
Leydig Cell ◽  
Half Life ◽  
Leydig Cells ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protein Band ◽  
Specific Protein ◽  
Rat Testis

The effect of luteinizing hormone (luteotropin) and cycloheximide on specific protein synthesis in rat testis Leydig cells has been investigated. Proteins were labelled with either I114C]leucine, [3H]leucine or [35S]methionine during incubation with Leydig-cell suspensions in vitro. Total protein was extracted from the cells and separated by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. No detectable increase in the synthesis of specific proteins could be observed after incubation of Leydig cells with luteinizing hormone for up to 1 h. However, after a 2h incubation period, an increase in [35S]methionine incorporation was observed in a protein with an apparent mol.wt. of 21000 (referred to as ‘protein 21’). When, after labelling of this protein with [35S]-methionine, Leydig cells were incubated for another 30min with cycloheximide, no decrease in radioactivity of this protein band was observed, indicating that it does not have a short half-life. However, another protein band was detected, which after incubation with cycloheximide disappeared rapidly, the reaction following first-order kinetics, with a half-life of about 11 min. This protein, with an apparent mol.wt. of 33000 (referred to as “protein 33”), was found to be located in the particulate fraction of the Leydig cell, and could not be demonstrated in other rat testis-cell types or blood cells. No effect of luteinizing hormone on molecular weight, subcellular localization or half-life of protein 33 was observed. A possible role for protein 33 and protein 21 in the mechanism of action of luteinizing hormone on testosterone production of Leydig cells is discussed.

scholarly journals The Effect of Cryptorchidism on the Quantitative Histology, Histochemistry and Hydrolytic Enzyme Activity of the Rat Testis

1978 ◽  
Vol 31 (1) ◽  
pp. 53 ◽  
Author(s):  
AW Blackshaw ◽  
PF Massey
Keyword(s):  
Enzyme Activity ◽  
Meiotic Prophase ◽  
Hydrolytic Enzyme ◽  
Seminiferous Tubules ◽  
Quantitative Histology ◽  
Rat Testis ◽  
Germinal Cells ◽  
Enzyme Patterns

Cryptorchidism of the mature rat testis led to degeneration of the seminiferous tubules and changes in enzyme patterns and activities. Spermatogenic stages 1--4, containing pachytene primary spermatocytes in late meiotic prophase, and stage 5, containing recently formed round spermatids, were damaged by 48 h. Within 96 h stages showed a loss of germinal cells into the lumen and this was almost complete by 192 h.

Interstitial fluid volume in the rat testis: androgen-dependent regulation by the seminiferous tubules?

1989 ◽  
Vol 120 (2) ◽  
pp. 215-NP ◽  
Author(s):  
S. Maddocks ◽  
R. M. Sharpe
Keyword(s):  
Leydig Cells ◽  
Interstitial Fluid ◽  
Single Injection ◽  
Fluid Volume ◽  
Male Rats ◽  
Seminiferous Tubules ◽  
Rat Testis ◽  
Adult Rat ◽  
Interstitial Fluid Volume ◽  
Dependent Mechanism

ABSTRACT Regulation of testicular interstitial fluid (IF) volume has been investigated in adult male rats in which the Leydig cells were selectively destroyed with a single i.p. injection of ethane dimethane sulphonate (EDS). Following this treatment, some animals also received testosterone supplementation by s.c. injection every 3 days, beginning either from the time of EDS injection, or 3–12 days afterwards. The volume of IF obtained by drip collection was determined, and testosterone and gonadotrophin concentrations measured in blood and in IF. Testosterone levels in IF and serum became undetectable by 3 days after EDS treatment. IF volume was reduced by 50% (P < 0·01) to reach a minimum level between 6 and 9 days after treatment. However, this decline was prevented in the absence of Leydig cells by supplementation with testosterone from the time of EDS injection, a treatment which also kept gonadotrophins at minimum or undetectable levels. Furthermore, the reduced IF volume seen up to 9 days after treatment with EDS alone could be restored to control levels within 3 days by a single injection of testosterone. The results obtained demonstrate that androgens, but not Leydig cells or gonadotrophins, are required for the maintenance of interstitial fluid volume in the adult rat testis. It is suggested that the seminiferous tubules may mediate this response, through an androgen-dependent mechanism. Journal of Endocrinology (1989) 120, 215–222

scholarly journals Leydig cell apoptosis in the rat testes after administration of the cytotoxin ethane dimethanesulphonate: role of the Bcl-2 family members

1998 ◽  
Vol 157 (2) ◽  
pp. 317-326 ◽  
Author(s):  
MF Taylor ◽  
I Woolveridge ◽  
AD Metcalfe ◽  
CH Streuli ◽  
JA Hickman ◽  
...  
Keyword(s):  
Cell Death ◽  
Leydig Cell ◽  
Leydig Cells ◽  
Family Members ◽  
Tumour Suppressor Gene ◽  
Male Rats ◽  
Seminiferous Tubules ◽  
Gene Products ◽  
Bax Protein ◽  
Ethane Dimethanesulphonate

Ethane dimethanesulphonate (EDS) is cytotoxic to Leydig cells in the adult rat. To investigate the role and regulation of apoptosis in the Leydig cell, EDS (100 mg/kg i.p.) was administered to adult male rats and the testes examined 6, 12, 18, 24, 48 and 72 h later. Numbers of Leydig cells, identified by 3 beta-hydroxysteroid dehydrogenase immuno-histochemistry started to fall by 12 h after EDS injection and were almost undetectable by 72 h. Apoptotic cells in the interstitium, visualised by in situ end labelling of DNA, increased in number to reach a maximum 24 h after injection of EDS, and were undetectable by 72 h. In many tissues the apoptosis-related gene products act in cohort: Bcl-2 and Bcl-xl promoting survival of a cell, whilst Bax promotes cell death often positively regulated by the tumour-suppressor gene p53. Western blot analysis showed that: (1) Bcl-2 and p53 were absent from interstitial Leydig cells but were expressed in the seminiferous tubules. (2) Bax protein although expressed in the interstitium was not present in the Leydig cells. (3) Bcl-xl in Leydig cells was transiently increased after EDS. In conclusion, EDS kills Leydig cells by apoptosis; however the control of Leydig cell death does not involve p53 or the Bcl-2 family members but may require other gene products yet to be identified.

Ethan-1,2-dimethanesulphonate reduces testicular oxytocin content and seminiferous tubule movements in the rat

1987 ◽  
Vol 112 (2) ◽  
pp. 311-NP ◽  
Author(s):  
H. D. Nicholson ◽  
R. T. S. Worley ◽  
S. E. F. Guldenaar ◽  
B. T. Pickering
Keyword(s):  
Leydig Cell ◽  
Seminiferous Tubule ◽  
Leydig Cells ◽  
Contractile Activity ◽  
Histological Study ◽  
Interstitial Cells ◽  
Seminiferous Tubules ◽  
Adult Rats ◽  
Adult Rat

ABSTRACT An oxytocin-like peptide is present in the interstitial cells of the testis, and testicular concentrations of oxytocin have been shown to increase seminiferous tubule movements in vitro. We have used the drug ethan-1,2-dimethanesulphonate (EDS), which depletes the Leydig cell population of the adult rat testis, to examine further the relationships between the Leydig cell, testicular oxytocin and tubular movements. Adult rats were injected i.p. with a single dose of EDS (75 mg/kg) or of vehicle (25% dimethyl sulphoxide). Histological study 3 and 10 days after treatment with EDS showed a reduction in the number of interstitial cells, and levels of oxytocin immunoreactivity were undetectable by radioimmunoassay. Immunostaining revealed very few oxytocin-reactive cells. Spontaneous contractile activity of the seminiferous tubules in vitro was also dramatically reduced, but could be restored by the addition of oxytocin to the medium. Four weeks after EDS treatment, the interstitial cells were similar to those in the control animals both in number and in immunostaining; immunoassayable oxytocin was present and tubular movements were normal. The EDS effect, seen at 3 and 10 days, was not altered by daily treatment with testosterone. However, repopulation of the testes with oxytocin-immunoreactive cells was not seen until 6 weeks in the testosterone-treated animals. We suggest that the Leydig cells are the main source of oxytocin immunoreactivity in the testis and that this oxytocin is involved in modulating seminiferous tubule movements and the resultant sperm transport. The results also imply that testosterone does not play a major role in controlling tubular activity in the mature rat. J. Endocr. (1987) 112, 311–316

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