Re-evaluation of the intratesticular level of testosterone required for quantitative maintenance of spermatogenesis in the rat

1988 ◽  
Vol 117 (1) ◽  
pp. 19-NP ◽  
Author(s):  
R. M. Sharpe ◽  
K. Donachie ◽  
I. Cooper

ABSTRACT The amount of testosterone required for quantitative maintenance of spermatogenesis has been re-evaluated using techniques aimed at minimizing the synthesis of testosterone after removal of the testis. Adult male rats were treated with ethane dimethane-sulphonate (EDS) to destroy the Leydig cells, and were supplemented with 25, 5 or 1 mg testosterone esters by injection every 3 days for 21 days. Serum hormone levels, testicular morphology and spermatogenesis were assessed and the intratesticular levels of testosterone compared in testes either removed under ether anaesthesia and placed in liquid nitrogen (right testis) or removed after collection of blood and placed in ice (left testis). Data for testosterone-supplemented rats were compared with those for control rats and rats treated with EDS alone. All doses of testosterone suppressed LH and FSH levels in serum to within the hypophysectomized range, and Leydig cell regeneration in EDS-treated rats was prevented completely. Treatment of EDS-injected rats with 25 or 5 mg testosterone maintained testicular weight, the number of germ cells and the diameter of seminiferous tubules at stage VII within or above the control range, although there was a significant increase in the number of degenerating pachytene spermatocytes at stage VII with 5 but not 25 mg testosterone; none of these parameters was maintained at control levels by a dose of 1 mg testosterone. Levels of testosterone in testosterone-supplemented rats differed little between testes collected in ice and liquid nitrogen, but in controls and rats treated with EDS alone, testosterone levels were overestimated by 75 and 27% respectively when comparing testes collected in ice with those collected in liquid nitrogen. This suggests that synthesis of testosterone does occur after removal and cooling of the testes. Using the data for testes collected in liquid nitrogen, the present results suggest that intratesticular levels of testosterone need to be maintained at 24–46% of control values (i.e. higher than previously shown) for quantitative maintenance of spermatogenesis, although it is argued that even these values probably still represent underestimates. J. Endocr. (1988) 117, 19–26

1995 ◽  
Vol 147 (3) ◽  
pp. 463-471 ◽  
Author(s):  
F Gaytan ◽  
C Bellido ◽  
C Morales ◽  
N van Rooijen ◽  
E Aguilar

Abstract The Leydig cells of young hypophysectomized rats are highly sensitive to the stimulatory effects of exogenous pituitary hormones. The aim of this study was to analyse the role of testicular macrophages in the response of Leydig cells to different hormones. Male rats were hypophysectomized at 28 days of age and 10 days later they were injected intratesticularly with dichloromethylene diphosphonate-containing liposomes (right testis) to deplete testicular macrophages, and with 0·9% NaCl (left testis). One week later, the animals were treated daily with 1 IU rat GH (rGH)/rat, 5 IU recombinant human FSH (recFSH)/rat, 10 IU human chorionic gonadotrophin (hCG)/rat, or vehicle for 7 days. The animals were killed on the day after the last injection. The animals treated with rGH showed increased body weight and increased number and size of testicular macrophages in the left testes, but no significant effects on Leydig cells were found. Treatment with recFSH induced a significant increase in testicular weight and tubular diameter in both testes. In the left testes, the number and size of macrophages were increased; the number of Leydig cells was not changed, although they showed a significantly increased cross-sectional area. This effect was abolished in the right (macrophage-depleted) testes. However, the effect of recFSH on the growth of the seminiferous tubules was not modified by the absence of macrophages. Rats treated with hCG showed increased testicular weight and serum testosterone levels, as well as an increased weight of the ventral prostate. In the left testes, the number and size of both macrophages and Leydig cells were increased. Otherwise, the number of Leydig cells was unchanged in the absence of macrophages, whereas the increase in the size of Leydig cells was partially abolished. These data indicate that testicular macrophages are needed for the response of Leydig cells to gonadotrophin treatment. Journal of Endocrinology (1995) 147, 463–471


1989 ◽  
Vol 37 (11) ◽  
pp. 1667-1673 ◽  
Author(s):  
R Schulz ◽  
F Paris ◽  
P Lembke ◽  
V Blüm

Androgens drive spermatogenesis by processes that are largely unknown. Direct effects on germ cells and indirect effects mediated via testicular somatic elements are currently under consideration, and specific localization of androgens in seminiferous tubules may provide information as regards this. Adult male rats were injected with ethane dimethanesulfonate (EDS; 75 mg/kg body weight) or vehicle. Testes were fixed and paraffin-embedded for localization of testosterone immunoreactivity 1 and 2 weeks after treatment, using the unlabeled antibody (PAP) technique. Plasma testosterone dropped from a pre-treatment level of 2.3 ng/ml to below 0.2 ng/ml 3 days after EDS injection and remained at low levels until the end of observation, accompanied by a progressive decrease in testicular weight. In the seminiferous tubules of vehicle-injected males, testosterone immunoreactivity was found in nuclei of spermatocytes and spermatids and in nuclei and the cytoplasm of Sertoli cells, and showed typical variations according to the stage of spermatogenesis. One week after EDS treatment, immunoreactivity had disappeared from the seminiferous epithelium. Two weeks after treatment, staining of germ cells was detected in two out of four males. The disappearance and reappearance of immunoreactivity coincided with the time course of EDS effects on rat Leydig cells, and we conclude that it corresponds to androgen specifically localized in fixed, paraffin-embedded tissue. Because staining of germ cell nuclei varied with the stage of spermatogenesis, the technique may detect a physiologically relevant androgen fraction; its location suggests that androgens may also directly affect certain germ cell stages.


1964 ◽  
Vol 46 (1) ◽  
pp. 25-30 ◽  
Author(s):  
Manuel Maqueo ◽  
Fred A. Kind

ABSTRACT Oestradiol-17β benzoate, 120 βg, injected into five-day old male rats inhibited maturation of the seminiferous epithelium as demonstrated by histological studies performed 40–55 days post-treatment. The oestrogen treatment was ineffective when administered at the age of 20 days. The degree of testicular damage appeared to be correlated with the amount of steroid used. A dose of 240 μg of oestradiol benzoate led to severe pathological changes in almost 100 per cent of the seminiferous tubules and atrophy of the Leydig cells.


2020 ◽  
Vol 13 (2) ◽  
pp. 125-136
Author(s):  
Mohammad Alwan ◽  
Baraa Al-Okialy

The present study was aimed to investigate the role of alpha-lipoic acid ( ALA) as an antioxidant against in ameliorating histological disorders of pituitary- testicular axis- induced by hydrogen peroxide (H2O2) in adult Wistar rats. Forty adult male rats were randomly divided into four equal groups (10 rats /group) and were handled daily as follows for 56 days: Control group ( C) were intubated distilled water and received ordinary tap water; group T1 were intubated 60mg/kg B.W of ALA and received ordinary tap water; group T2 were administered H2O2 in tap water at a concentration of 0.05%, while group T3 were intubated 60mg/kg B.W of ALA and received ordinary tap water containing 0.05% H2O2. At the end of the experiment, body weights were recorded, then pituitary and testes were excised for histopathological study and testicular weight was recorded too. Rats administered H2O2 showed a significant decrease in testes weight to body weight ratio accompanied with major histopathological changes of the testes in comparison with other groups including; a significant decrease in the diameter of seminiferous tubules, high of germinal epithelial cell and degenerative changes with incomplete spermatogenesis. Besides, a significant decrease in the number of Leydig's cells in comparison with other experimental groups. Furthermore, pituitary gland of group T2 manifested a severe histological alteration in architecture characterized by atrophy with marked necrotic and degenerative changes. Whereas, rats administered ALA (group T3) shows an improvement of histological changes of pituitary and testicular tissues induced by hydrogen peroxide. In conclusion, the results indicated that alpha-lipoic acid mitigated pituitary-testicular dysfunctions induced by H2O2 through its antioxidant effects via scavenging free radicals.


2021 ◽  
Vol 10 (1) ◽  
pp. 18
Author(s):  
Kukuh Prastyaningtyas ◽  
Rochmah Kurnijasanti ◽  
Rahmi Sugihartuti ◽  
Suherni Susilowati ◽  
Tri Wahyu Suprayogi ◽  
...  

This study aims to determine the effect of red dragon (Hylocereus polyrhizus) fruit peel extract (RDFPE) on the parameters of Leydig cells number, seminiferous tubules diameter, and testicular weight of mice (Mus musculus) exposed to heat (40°C). Twenty adult male mice were divided randomly into five groups. The control group (C) mice only received a placebo. Meanwhile, the treatment groups mice were exposed to heat for 45 minutes daily for 36 days and oral administration of placebo, RDFPE of 250, 500, and 1000mg/kg BW for T0, T1, T2, and T3, respectively. The result showed that heat exposure on mice (T0 group) caused a lower of all of the parameters (p <0.05) than normal mice (control group, C). RDFPE administration at a dose of 250 mg/kg BW (T1 group) and 500 mg/kg BW (T2 group) resulted in a higher value of those parameters (p <0.05) compared to the T0 group. All those parameters of the T2 group (dose of 500 mg/kg BW) were not significantly different (p >0.05) than the control group (normal mice). However, the higher dose of RDFPE (1000 mg/kg BW, T3 group) resulted in the lower values of those parameters (p <0.05) than those of the T2 group. It could be concluded that 500mg/kg BW dose of RDFPE could return Leydig cells number, seminiferous tubules diameter, and testicular weight of mice (Mus musculus) exposed to heat.


1989 ◽  
Vol 120 (2) ◽  
pp. 215-NP ◽  
Author(s):  
S. Maddocks ◽  
R. M. Sharpe

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


1998 ◽  
Vol 157 (2) ◽  
pp. 317-326 ◽  
Author(s):  
MF Taylor ◽  
I Woolveridge ◽  
AD Metcalfe ◽  
CH Streuli ◽  
JA Hickman ◽  
...  

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.


2016 ◽  
Vol 3 (2) ◽  
Author(s):  
Alicia González-González ◽  
Everardo González-Padilla ◽  
Francisco Fierro-Fierro ◽  
María De Lourdes Juárez-Mosqueda ◽  
Juan José Pérez-Rivero ◽  
...  

The aim of this study was to determine the effect of tamoxifen (Tx) and its vehicle, soybean oil (SO), during the critical period of hypothalamic sexual differentiation in newborn male rats, regarding gonadal histomorphology during adulthood. The animals were randomly divided into 3 groups (n = 5 each). An hour after birth, one group was treated subcutaneously with 200 μg of Tx, using commercial SO (20 uL) as a vehicle; another group was treated with only 20 μL of SO; the control group received no treatment. All rats were weighed and sacrificed by cervical dislocation on day 90 post-treatment. Testicles were removed, weighed and processed for histological evaluation. The single administration of Tx and/or SO during the critical period of sexual differentiation of the hypothalamus permanently altered testicular histomorphology, spermatogenesis, and body weight in adulthood. Alterations included vacuolization and reduction in the number of spermatogonia and Sertoli cells. The administration of Tx reduced the testicular weight, the diameter and area of the seminiferous tubules, and the height of the germinal epithelium, and increased the intertubular space. Soybean oil by itself reduced the number of spermatocytes and spermatids more than Tx did. There was no effect on the number of Leydig cells. The possibility that soybean oil can act as an endocrine disruptor deserves greater attention and opens the possibility for the development of new methods of pest control.


2001 ◽  
Vol 20 (8) ◽  
pp. 393-397 ◽  
Author(s):  
K Ben Rhouma ◽  
O Tébourbi ◽  
R Krichah ◽  
M Sakly

The reproductive toxicity of DDT was investigated in adult male rats exposed to 50 and 100 mg/kg body weight (b.wt) day 1 for 10 successive days. Compared with control animals, administration of DDT led to a dose-dependent reduction of testicular weight and the number as well as the percentage of motile spermatozoa in the epididymis. Testicular histological observationsrevealed alsoamarkedloss of gametes in the lumen of seminiferous tubules. In DDT treated rats, the seminal vesicles weights dropped significantly, resulting from a decrease of testosterone production by testes, whereas serum LH and FSH increased after pesticide exposure. This increase of gonadotrophin levels may be related to an impairment of the negative feedback exerted by the steroid on the hypothalamic–pituitary axis. It is concluded that DDT induced adverse effects on male rat fertility by acting directly on the testes and altering the neuroendocrinefunction.


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