Testicular steroidogenesis in the testicular feminized (Tfm) mouse: loss of 17α-hydroxylase activity

1991 ◽  
Vol 131 (3) ◽  
pp. 443-449 ◽  
Author(s):  
L. Murphy ◽  
P. J. O'Shaughnessy
Keyword(s):  
Leydig Cells ◽  
Smooth Endoplasmic Reticulum ◽  
Reductase Activity ◽  
Serum Testosterone ◽  
Normal Activity ◽  
Testicular Descent ◽  
Hydroxylase Activity ◽  
Steroid Production ◽  
And Function

ABSTRACT Testicular feminized (Tfm) mice are totally insensitive to androgen and may be used to study the role of the androgen receptor in normal development and function. We have examined testicular and Leydig cell steroidogenesis in Tfm mice. Serum bioactive LH was high in Tfm mice but serum testosterone was low and this was associated with a severe reduction in testicular testosterone production in vitro. Examination of [3H]pregnenolone metabolism by testes of Tfm mice indicated that progesterone, rather than testosterone, was the major steroid produced. Leydig cells were isolated from normal and Tfm mice and from normal mice in which testicular descent was surgically prevented before puberty. As in whole testes, androgen production in response to human chorionic gonadotrophin was severely reduced in Leydig cells from testes of Tfm mice compared with normal or cryptorchid groups. In contrast, progesterone production by Leydig cells from testes of Tfm mice was markedly increased in comparison with other groups. Total steroid production (progesterone plus androstenedione plus testosterone), however, was only 24% of normal in Leydig cells from Tfm mice. The pattern of steroid production by Leydig cells from cryptorchid testes was similar to control, although total steroid production was reduced to about 50% (this was significantly higher than the Tfm group, P<0·05). The high progesterone/androgen ratio in testes from Tfm mice suggested that 17α-hydroxylase was depleted in these animals. To confirm this, activity of the four major steroidogenic enzymes associated with the smooth endoplasmic reticulum was measured. Activities (per testis) of 3β-hydroxysteroid dehydrogenase and 5α-reductase were normal in Tfm and cryptorchid mice but, as expected, 17α-hydroxylase activity was only 2·4% of control and 4·5% of cryptorchid testes. 17-Ketosteroid reductase activity was markedly reduced in cryptorchid testes (14·4% of control) but there was a further reduction in testes from Tfm mice to 0·1% of control. Results show that the Tfm mutation is associated with marked loss of 17α-hydroxylase and 17-ketosteroid reductase activities. This suggests that these enzymes may require receptor-mediated androgen stimulation during development to express normal activity. Journal of Endocrinology (1991) 131, 443–449

The influence of luteinizing hormone on the Leydig cells of cryptorchid rat testes

1984 ◽  
Vol 107 (1) ◽  
pp. 110-116 ◽  
Author(s):  
L.J. Wilton ◽  
D. M. de Kretser
Keyword(s):  
Cell Size ◽  
Leydig Cell ◽  
Leydig Cells ◽  
Smooth Endoplasmic Reticulum ◽  
Serum Testosterone ◽  
Pituitary Hormones ◽  
Adult Rats ◽  
Serum Hormone ◽  
Local Feedback

Abstract. The influence of circulating LH levels on Leydig cells from cryptorchid adult rats was examined after ablation of the pituitary. After 2 weeks cryptorchidism, serum FSH and LH levels rose 2-fold while serum testosterone (T) remained unchanged. Leydig cells were hypertrophied and showed an increased response to in vitro hCG stimulation. Two weeks after hypophysectomy (hypox), serum hormone levels (LH, FSH and T), Leydig cell size, cytoplasm, organelle content and in vitro T production were all dramatically reduced. However, when hypophysectomy was combined with cryptorchidism (hypox/crypt), there was an increase in Leydig cell size, compared to hypophysectomy alone, in the presence of very low levels of serum FSH, LH and T. Compared to the hypophysectomised state, the mitochondria were larger and the cytoplasm contained more smooth endoplasmic reticulum. The response of the hypox/crypt testes to in vitro hCG stimulation, though significantly less than the cryptorchid testes, was significantly greater than the hypox testes. These results demonstrate that the changes observed in the Leydig cell after cryptorchidism can occur in the absence of peripheral pituitary hormones and are consistent with the hypothesis that a local feedback loop exists within the testis.

scholarly journals Isolation and characterization of Leydig cells from adult bonnet monkeys (Macaca radiata): evidence for low steroidogenic capacity in monkey Leydig cells in contrast to rat Leydig cells

2003 ◽  
Vol 179 (2) ◽  
pp. 175-182 ◽  
Author(s):  
M Anbalagan ◽  
V Sriraman ◽  
A Jagannadha Rao
Keyword(s):  
Leydig Cells ◽  
Smooth Endoplasmic Reticulum ◽  
Macaca Radiata ◽  
Enzymatic Dissociation ◽  
Isolation And Characterization ◽  
Chain Cleavage ◽  
Cleavage Enzyme ◽  
Gradient Fractionation ◽  
Available Information

Most of the available information on Leydig cells has been obtained using a rodent model system. With an objective to extend the observations made with rat Leydig cells (RLCs) to primates, a method has been developed to isolate Leydig cells from monkey (Macaca radiata) testis. Enzymatic dissociation of monkey testis followed by Percoll-gradient fractionation of the interstitial cells resulted in the recovery of Leydig cells at densities corresponding to 1.064-1.070 g/ml. Purified (90-94%) monkey Leydig cells (MLCs) stained positive for the Leydig cell marker 3beta-hydroxysteroid dehydrogenase. The cells responded to in vitro addition of human chorionic gonadotropin (hCG) and produced testosterone. Comparison of the in vitro testosterone-producing ability of MLCs with RLCs revealed that MLCs have much less steroidogenic capacity compared with the RLCs. Analysis revealed that limitation in substrate availability to mitochondrial P(450) side chain cleavage enzyme and low mitochondrial and smooth endoplasmic reticulum content in MLCs could be the possible reasons for the low steroidogenic capacity of the MLCs.

scholarly journals Evaluation of relative roles of LH and FSH in regulation of differentiation of Leydig cells using an ethane 1,2-dimethylsulfonate-treated adult rat model

2003 ◽  
Vol 176 (1) ◽  
pp. 151-161 ◽  
Author(s):  
V Sriraman ◽  
MR Sairam ◽  
AJ Rao
Keyword(s):  
Leydig Cells ◽  
Serum Testosterone ◽  
Side Chain ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Lh Receptor ◽  
Side Chain Cleavage ◽  
Chain Cleavage ◽  
Cleavage Enzyme

The relative role of LH and FSH in regulation of differentiation of Leydig cells was assessed using an ethane 1,2-dimethylsulfonate (EDS)-treated rat model in which endogenous LH or FSH was neutralized from day 3 to day 22 following EDS treatment. Serum testosterone and the in vitro response of the purified Leydig cells to human chorionic gonadotropin (hCG) was monitored. In addition RNA was isolated from the Leydig cells to monitor the steady-state mRNA levels by RT-PCR for 17alpha-hydroxylase, side chain cleavage enzyme, steroidogenic acute regulatory protein (StAR), LH receptor, estrogen receptor (ER-alpha) and cyclophilin (internal control). Serum testosterone was undetected and the isolated Leydig cells secreted negligible amount of testosterone on stimulation with hCG in the group of rats that were treated with LH antiserum following EDS treatment. RT-PCR analysis revealed the absence of message for cholesterol side chain cleavage enzyme and 17alpha-hydroxylase although ER-alpha and LH receptor mRNA could be detected, indicating the presence of undifferentiated precursor Leydig cells. In contrast, the effects following deprival of endogenous FSH were not as drastic as seen following LH neutralization. Deprival of endogenous FSH in EDS-treated rats led to a significant decrease in serum testosterone and in vitro response to hCG by the Leydig cells. Also, there was a significant decrease in the steady-state mRNA levels of 17alpha-hydroxylase, cholesterol side chain cleavage enzyme, LH receptor and StAR as assessed by a semiquantitative RT-PCR. These results establish that while LH is obligatory for the functional differentiation of Leydig cells, repopulation of precursor Leydig cells is independent of LH, and also unequivocally establish an important role for FSH in regulation of Leydig cell function.

scholarly journals Glucocorticoids Affect Male Testicular Steroidogenesis

2020 ◽  
pp. S205-S210
Author(s):  
R. HAMPL ◽  
L. STÁRKA
Keyword(s):  
Leydig Cells ◽  
Glucocorticoid Receptors ◽  
Molecular Mechanisms ◽  
Target Cells ◽  
Clinical Effects ◽  
Steroid Production ◽  
Glucocorticoid Treatment ◽  
Testicular Steroidogenesis ◽  
And Function

Through their receptors at each level of hypothalamo-pituitary-gonadal axis glucocorticoid excess, either endogenous or administered or stress-induced, could affect steroid production in the testis and thus male fertility. The main ways by which glucocorticoids act are as follows: 1) Affecting gonadoliberin and LH synthesis and release through glucocorticoid receptors in hypothalamic neurons and pituitary gonadotropes. 2) By so far not clearly evidenced reduction of the number of LH receptors on the membrane of Leydig cells. 3) By affecting expression and function of steroidogenic enzymes in the testis. 4) By regulation of in situ access of glucocorticoid to its target cells in the testis. 5) By promotion Leydig cell apoptosis. The review provides a survey of physiological and molecular mechanisms staying behind these effects. It does not deal with the clinical effects of glucocorticoid treatment which would substantially exceed the scope of the pater.

EFFECTS OF STEROID SUPPRESSION AND GONADECTOMY ON ADRENAL 5α-REDUCTASE ACTIVITY AND CORTICOSTERONE PRODUCTION IN RATS

1973 ◽  
Vol 74 (3) ◽  
pp. 568-575 ◽  
Author(s):  
Howard D. Colby ◽  
Raphael J. Witorsch ◽  
James L. Caffrey ◽  
Julian I. Kitay
Keyword(s):  
Reductase Activity ◽  
Gonadal Hormones ◽  
Adrenocortical Function ◽  
Common Site ◽  
Steroid Production ◽  
Adult Females ◽  
Site Of Action ◽  
Before And After ◽  
Intact Rats

ABSTRACT Studies were conducted to further examine the interaction between gonadal hormones and ACTH in the regulation of adrenocortical function. Gonadectomy for a period of two weeks in pre-puberal rats of either sex or in adults for one week, was without effect on adrenal 5α-reductase activity and corticosterone production in vitro. Cortisone administration to intact rats, both before and after puberty, suppressed corticosterone and total steroid production and increased adrenal reductase activity. Ovariectomy in immature or adult females did not modify the effects of cortisone on adrenal function. Orchiectomy, both before and after puberty, synergistically enhanced the actions of cortisone to increase 5α-reductase activity and lower corticosterone production without affecting total steroid output. These observations lend support to the hypothesis that gonadal hormones interact with ACTH in the control of adrenocortical function. Adrenal 5α-reductase seems to be a common site of action for the effects.

Accelerated recovery of Leydig cells of the immature rat testis after administration of the cytotoxic ethylene dimethanesulphonate

1988 ◽  
Vol 119 (3) ◽  
pp. 475-NP ◽  
Author(s):  
G. Edwards ◽  
R. G. Lendon ◽  
I. D. Morris
Keyword(s):  
Leydig Cells ◽  
Serum Testosterone ◽  
Hydroxysteroid Dehydrogenase ◽  
Cytotoxic Action ◽  
Immature Rat ◽  
Control Value ◽  
Adult Rat ◽  
Cytocidal Effect ◽  
Old Rats

ABSTRACT A single injection of ethane-1,2-dimethanesulphonate (EDS; 100 mg/kg) selectively destroys Leydig cells in the testis of the adult rat; however, unconfirmed reports indicate that Leydig cells in the immature rat are not affected. In this study the effect of EDS was examined 2 days after treatment of rats aged 20, 25 or 35 days. There was a large reduction in the in-vitro binding of 125I-labelled human chorionic gonadotrophin (hCG) to the homogenates of testes from EDS-treated immature rats. EDS reduced the testosterone content of the testes at all ages studied, but 2 days after injection had only significantly lowered the serum testosterone concentration of 25- or 35-day-old animals. Light microscopic examination of the testis of the 22-day-old rat, 2 days after treatment with EDS, indicated that there were still many cells staining for 3β-hydroxysteroid dehydrogenase. The interstitium also contained numerous atypical cells which did not stain for 3β-hydroxysteroid dehydrogenase. Electron microscopy of testes from the 22-day-old EDS-treated rat showed that Leydig cells were still present in the interstitium together with macrophages and fibroblast-like cells. Six days after EDS treatment of 20-day-old rats, but not 35-day-old rats, there was an increase in the binding of 125I-labelled hCG to testis homogenate to 70% of control value. Testicular testosterone content 6 days after treatment of the 20-day-old rat had risen to 50% of the control testis value. These changes documented in the 20-day-old rat after EDS treatment can be explained by either a cytocidal effect with subsequent repopulation of new Leydig cells which has been described in the adult rat or by a reversible cytotoxic action which has not previously been documented. J. Endocr. (1988) 119, 475–482

Inhibition of 17β-hydroxysteroid-oxidoreductase activity in purified rat Leydig cells by single injection of human choriogonadotrophin

1984 ◽  
Vol 105 (4) ◽  
pp. 571-576 ◽  
Author(s):  
Nikolaus Kühn-Velten ◽  
Wolfgang Staib
Keyword(s):  
Leydig Cells ◽  
Time Course ◽  
Marked Decrease ◽  
Single Injection ◽  
Reductase Activity ◽  
Competitive Inhibitor ◽  
Oxidoreductase Activity ◽  
Rate Limiting

Abstract. A marked decrease of progesterone conversion to androgens in vitro by purified rat testis Leydig cells is observed from 1/2 to 3 days after treatment of the rats with 100 IU human choriogonadotrophin (hCG) in vivo. The maximal inhibition results 2 days after hCG injection and is denoted by a 72% reduction of androstenedione formation and a 78% reduction of testosterone formation from 2.5 μmol progesterone · l7#x2212;1. The testosterone/androstenedione ratio (about 0.4), however, is not changed after hCG treatment, indicating that the 17-ketosteroid-reductase activity is not rate-limiting under these conditions. Nevertheless, testosterone formation from 2.5 μmol androstenedione · l−1 is reduced by 67% and androstenedione formation from 2.5 μmol testosterone · l−1 is reduced by 79% 1 and 2 days after hCG treatment. The time-course of this hCG-induced decrease in both reductase and oxidase activities of the Leydig cell 17β-hydroxysteroid-oxidoreductase parallels the decrease of androgen formation from progesterone. Kinetic analyses reveal that the Vmax of the oxidase is reduced to a significantly (P < 0.05) greater extent than the Vmax of the reductase (79 and 62%, respectively), whereas the respective Km values remain unchanged. From these results it may be concluded that either a loss, or an inactivation of the enzyme protein, or the formation of non-competitive inhibitor occur during hCG action and that hCG may affect different enzyme activities involved in testicular androgen biosynthesis in a similar way.

scholarly journals Monocyte Chemoattractant Protein-1 stimulates the differentiation of rat stem and progenitor Leydig cells during regeneration

2020 ◽  
Vol 20 (1) ◽  
Author(s):  
Xiangcheng Zhan ◽  
Jingwei Zhang ◽  
Saiyang Li ◽  
Xiaolu Zhang ◽  
Linchao Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Leydig Cell ◽  
Leydig Cells ◽  
Serum Testosterone ◽  
Monocyte Chemoattractant Protein 1 ◽  
Rat Testis ◽  
Monocyte Chemoattractant ◽  
Monocyte Chemoattractant Protein

Abstract Background Monocyte chemoattractant protein-1(MCP-1) is a chemokine secreted by Leydig cells and peritubular myoid cells in the rat testis. Its role in regulating the development of Leydig cells via autocrine and paracrine is still unclear. The objective of the current study was to investigate the effects of MCP-1 on Leydig cell regeneration from stem cells in vivo and on Leydig cell development in vitro. Results Intratesticular injection of MCP-1(10 ng/testis) into Leydig cell-depleted rat testis from post-EDS day 14 to 28 significantly increased serum testosterone and luteinizing hormone levels, up-regulated the expression of Leydig cell proteins, LHCGR, SCARB1, CYP11A1, HSD3B1, CYP17A1, and HSD17B3 without affecting progenitor Leydig cell proliferation, as well as increased ERK1/2 phosphorylation. MCP-1 (100 ng/ml) significantly increased medium testosterone levels and up-regulated LHCGR, CYP11A1, and HSD3B1 expression without affecting EdU incorporation into stem cells after in vitro culture for 7 days. RS102895, a CCR2 inhibitor, reversed MCP-1-mediated increase of testosterone level after culture in combination with MCP-1. Conclusion MCP-1 stimulates the differentiation of stem and progenitor Leydig cells without affecting their proliferation.

Zebrafish cyp11c1 Knockout Reveals the Roles of 11-ketotestosterone and Cortisol in Sexual Development and Reproduction

Endocrinology ◽  
2020 ◽  
Vol 161 (6) ◽  
Author(s):  
Qifeng Zhang ◽  
Ding Ye ◽  
Houpeng Wang ◽  
Yaqing Wang ◽  
Wei Hu ◽  
...  
Keyword(s):  
Sexual Development ◽  
Leydig Cells ◽  
Sex Differentiation ◽  
Germinal Vesicle ◽  
Genital Papilla ◽  
Germinal Vesicle Breakdown ◽  
And Function ◽  
Defective Spermatogenesis

Abstract Androgen is essential for male development and cortisol is involved in reproduction in fishes. However, the in vivo roles of cortisol and specific androgens such as 11-ketotestosterone (11-KT) in reproductive development need to be described with genetic models. Zebrafish cyp11c1 encodes 11β-hydroxylase, which is essential for the biosynthesis of 11-KT and cortisol. In this study, we generated a zebrafish mutant of cyp11c1 (cyp11c1-/-) and utilized it to clarify the roles of 11-KT and cortisol in sexual development and reproduction. The cyp11c1-/- fish had smaller genital papilla and exhibited defective natural mating but possessed mature gametes and were found at a sex ratio comparable to the wildtype control. The cyp11c1-/- males showed delayed and prolonged juvenile ovary-to-testis transition and displayed defective spermatogenesis at adult stage, which could be rescued by treatment with 11-ketoandrostenedione (11-KA) at certain stages. Specifically, during testis development of cyp11c1-/- males, the expression of insl3, cyp17a1, and amh was significantly decreased, suggesting that 11-KT is essential for the development and function of Leydig cells and Sertoli cells. Further, spermatogenesis-related dmrt1 was subsequently downregulated, leading to insufficient spermatogenesis. The cyp11c1-/- females showed a reduction in egg spawning and a failure of in vitro germinal vesicle breakdown, which could be partially rescued by cortisol treatment. Taken together, our study reveals that zebrafish Cyp11c1 is not required for definite sex differentiation but is essential for juvenile ovary-to-testis transition, Leydig cell development, and spermatogenesis in males through 11-KT, and it is also involved in oocyte maturation and ovulation in females through cortisol.

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