Mumps Virus Decreases Testosterone Production and Gamma Interferon-Induced Protein 10 Secretion by Human Leydig Cells

ABSTRACT Mumps virus is responsible for sterility. Here, we show that the mumps virus infects Leydig cells in vitro and totally inhibits testosterone secretion and that ribavirin in mumps virus-infected Leydig cell cultures completely restores testosterone production. Moreover, we show that gamma interferon-induced protein 10 (IP-10) is highly expressed by mumps virus-infected Leydig cells and that ribavirin does not block IP-10 production.

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Increase in Leydig cell responsiveness in the unilaterally cryptorchid rat testis and its relationship to the intratesticular levels of testosterone

Journal of Endocrinology ◽

10.1677/joe.0.1020319 ◽

1984 ◽

Vol 102 (3) ◽

pp. 319-327 ◽

Cited By ~ 26

Author(s):

R. M. Sharpe ◽

I. Cooper ◽

D. G. Doogan

Keyword(s):

Leydig Cell ◽

Leydig Cells ◽

Venous Blood ◽

Cell Interaction ◽

Adult Rats ◽

Similar Reduction ◽

Testosterone Production ◽

Lh Releasing Hormone ◽

Unilateral Cryptorchidism

ABSTRACT Adult rats were made unilaterally cryptorchid (UCD) and 6–7 weeks later Leydig cells were isolated from the scrotal and abdominal testes and their capacity to secrete testosterone in vitro was compared. Basal testosterone production by Leydig cells from the abdominal testes of UCD rats was lowered, compared with cells from the contralateral scrotal testes, whilst their responsiveness to both human chorionic gonadotrophin and an LH releasing hormone agonist was enhanced two- to threefold (P< 0·001) compared both with cells from the contralateral scrotal testes and with cells isolated from untreated rats of the same age. In the UCD rats, concentrations of testosterone in testicular interstitial fluid (IF) were reduced (P< 0·001) by 70–90% in abdominal, compared with scrotal, testes. A similar reduction was evident in the levels of testosterone in spermatic venous blood, and both this decrease and that in IF levels of testosterone varied according to the degree of testicular involution. The ontogeny of the above changes was investigated. After induction of unilateral cryptorchidism, the weight of the abdominal compared with the scrotal testis declined slowly, such that by day 5 there was only a 25% reduction in weight compared with a 70% reduction by day 40. In contrast, the levels of testosterone in IF from abdominal testes declined rapidly, such that by day 5 an 80% reduction was attained, compared with scrotal testes, with little further change by day 40. Hormone-stimulated testosterone production by Leydig cells isolated from the abdominal testes was unchanged or marginally reduced over the first 3 days compared with cells from the scrotal testes, but by day 5 there was a significant increase in responsiveness; this increase was of smaller magnitude than that evident at day 40. These results suggest a possible association between the fall in intratesticular levels of testosterone induced by unilateral cryptorchidism and the Leydig cell hypertrophy and hyper-responsiveness that occurs in the same testes. The implications with respect to altered Sertoli–Leydig cell interaction are discussed. J. Endocr. (1984) 102, 319–327

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Effects of melatonin treatment on Leydig cell activity in the testis of the frog Rana esculenta

Zygote ◽

10.1017/s0967199404002898 ◽

2004 ◽

Vol 12 (4) ◽

pp. 293-299 ◽

Cited By ~ 12

Author(s):

Michela d'Istria ◽

Ismene Serino ◽

Gaia Izzo ◽

Diana Ferrara ◽

Gianluca De Rienzo ◽

...

Keyword(s):

Leydig Cell ◽

Leydig Cells ◽

Morphological Changes ◽

Cell Activity ◽

Rana Esculenta ◽

Cellular Adhesion ◽

Morphological Observation ◽

Melatonin Treatment ◽

Testosterone Secretion

This study was conducted to verify the effect(s) of melatonin treatment on frog Leydig cells. Morphological observation after melatonin treatment indicates that many frog Leydig cells show degenerative changes (i.e. heterochromatic nuclei, loss of cellular adhesion) while in adjacent germinal tubules several Sertoli cells show heterochromatic nuclei, confirming the presence of a paracrine effect between interstitial and germinal compartments. The effect of melatonin on frog Leydig cell steroidogenesis was investigated in in vitro experiments; after 6 h of incubation melatonin severely inhibits both control and GnRH-induced testosterone secretion. In addition, in order to verify the effect of indolamine on frog Leydig cell activity, we investigated, by in situ hybridization, the presence of frog relaxin (fRLX, a transcript specifically expressed by these cells) in the testes of melatonin-injected animals after 48 h. fRLX signal completely disappeared from the testis of melatonin-injected frogs. The results of the present study indicate that melatonin treatment provokes Leydig cell morphological changes, blocks GnRH-antagonist-induced testosterone secretion and decreases fRLX expression. Taken together these results strongly indicate that melatonin acts on Leydig cells in the testis of the frog Rana esculenta.

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Effects of clodronate-containing liposomes on testicular macrophages and Leydig cells in vitro

Journal of Endocrinology ◽

10.1677/joe.0.1550087 ◽

1997 ◽

Vol 155 (1) ◽

pp. 87-92 ◽

Cited By ~ 7

Author(s):

DE Brigham ◽

G Little ◽

YO Lukyanenko ◽

JC Hutson

Keyword(s):

Direct Effect ◽

Dose Response ◽

Leydig Cell ◽

Leydig Cells ◽

Direct Effects ◽

Testosterone Production ◽

Identical Response ◽

Testicular Macrophages

We undertook the present studies to determine if clodronate-containing liposomes have direct effects on Leydig cells. Macrophages and Leydig cells were isolated and maintained separately in culture. Following treatment with clodronate-containing liposomes, macrophages were killed in a dose-response fashion over a range of 5-200 microliters liposomes. By comparison, a 500 microliters dose was required to kill Leydig cells, but this was not dependent upon clodronate since liposomes containing buffer elicited an identical response. The concentration of testosterone in medium from Leydig cells treated with clodronate-containing liposomes was significantly reduced compared with untreated cells. However, we subsequently found that liposomes can adsorb testosterone. Therefore, testosterone production was determined at various times following removal of liposomes from Leydig cells, thereby circumventing this complication. It was found that testosterone production was not altered by liposomes under these conditions. Finally, free clodronate had no effect on testosterone production, even at doses representing the amount present within the 500 microliters dose of liposomes. In summary, clodronate-containing liposomes killed testicular macrophages at a far smaller dose than required to kill Leydig cells. Most importantly, neither liposomes no free clodronate had a direct effect on testosterone production. Thus, clodronate-containing liposomes represent a valuable tool to study Leydig cell-macrophage interactions.

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Reversal of long-term LH deprivation on testosterone secretion and Leydig cell volume, number and proliferation in adult rats

Journal of Endocrinology ◽

10.1677/joe.0.1270047 ◽

1990 ◽

Vol 127 (1) ◽

pp. 47-NP ◽

Cited By ~ 25

Author(s):

D. S. Keeney ◽

R. L. Sprando ◽

B. Robaire ◽

B. R. Zirkin ◽

L. L. Ewing

Keyword(s):

Cell Volume ◽

Leydig Cell ◽

Leydig Cells ◽

Cell Number ◽

Implant Removal ◽

Adult Rats ◽

Testosterone Secretion ◽

Lh Secretion

ABSTRACT The purpose of this study was to determine whether Leydig cell volume and function could recover fully from long-term LH deprivation upon restoration of endogenous LH secretion, and whether the restoration of LH would elicit a mitogenic response, i.e. stimulate Leydig cell proliferation or affect Leydig cell number per testis. LH secretion was inhibited by treating adult rats with testosterone and oestradiol-filled (TO) silicone elastomer implants (16 weeks), and was restored by removing the implants. Changes in serum concentrations of LH and FSH, LH-stimulated testosterone secretion by testes perfused in vitro, Leydig cell volume and number per testis, average Leydig cell volume and Leydig cell [3H]thymidine incorporation were measured at weekly intervals following implant removal. The TO implants inhibited (P < 0·01) LH secretion, but serum concentrations of FSH were not significantly different (P > 0·10) from control values. After implant removal, serum LH returned to control values within 1 week, whereas serum FSH increased twofold (P < 0·01) and returned to control values at 4 weeks. LH-stimulated in-vitro testosterone secretion was inhibited by more than 99% in TO-implanted rats, but increased (P < 0·01) to 80% of control values by 8 weeks after implant removal. The total volume of Leydig cells per testis and the volume of an average Leydig cell were 14 and 19% of control values respectively, after 16 weeks of TO implantation (P < 0·01), but returned to 83 and 86% of controls (P > 0·10) respectively, by 6 weeks after implant removal. Leydig cell proliferation ([3H]thymidine labelling index) was low (< 0·1%) in both control and TO-implanted rats, increased (P < 0·01) fivefold from 1 to 4 weeks after implant removal and then declined to control values at 6 weeks. The increase in Leydig cell [3H]thymidine incorporation was mimicked by treating TO-implanted rats with exogenous LH, but not FSH. Leydig cells were identified in both the interstitium and the lamina propria of the seminiferous epithelium. The proportion of Leydig cell nuclei in the lamina propria was 30-fold greater (P < 0·01) at 1 and 3 weeks after implant removal (3%) compared with that for control and TO-implanted rats (0·1%). Total Leydig cell number per testis was marginally but not significantly (P = 0·06) decreased in rats treated with TO implants for 16 weeks when compared with controls (18·4±2·2 vs 25·4±1·2 × 106). Three weeks after implant removal, the numbers of Leydig cells per testis were identical (26·8±2·8 × 106) to those in control animals. These results not only demonstrate dramatic morphogenic effects of LH on mature rat Leydig cells, but also suggest that endogenous LH might be mitogenic at least to a subpopulation of Leydig cells. Journal of Endocrinology (1990) 127,47–58

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Adrenocorticotropic Hormone Directly Stimulates Testosterone Production by the Fetal and Neonatal Mouse Testis

Endocrinology ◽

10.1210/en.2003-0277 ◽

2003 ◽

Vol 144 (8) ◽

pp. 3279-3284 ◽

Cited By ~ 72

Author(s):

P. J. O’Shaughnessy ◽

L. M. Fleming ◽

G. Jackson ◽

U. Hochgeschwender ◽

P. Reed ◽

...

Keyword(s):

Leydig Cell ◽

Leydig Cells ◽

Cell Function ◽

Expressed Sequence Tag ◽

Acth Stimulation ◽

Testosterone Production ◽

Testicular Cells ◽

Testicular Steroidogenesis ◽

Fetal Leydig Cells

Abstract Adult Leydig cell steroidogenesis is dependent on LH but fetal Leydig cells can function independently of gonadotropin stimulation. To identify factors that may be involved in regulation of fetal Leydig cells expressed sequence tag libraries from fetal and adult testes were compared, and fetal-specific genes identified. The ACTH receptor [melanocortin type 2 receptor (Mc2r)] was identified within this fetal-specific group. Subsequent real-time PCR studies confirmed that Mc2r was expressed in the fetal testis at 100-fold higher levels than in the adult testis. Incubation of fetal or neonatal testes with ACTH in vitro stimulated testosterone production more than 10-fold, although ACTH had no effect on testes from animals aged 20 d or older. The steroidogenic response of fetal and neonatal testes to a maximally stimulating dose of human chorionic gonadotropin was similar to the response shown to ACTH. The ED50 for ACTH, measured in isolated fetal and neonatal testicular cells, was 5 × 10−10m and the lowest dose of ACTH eliciting a response was 2 × 10−11m. Circulating ACTH levels in fetal mice were around 8 × 10−11m. Neither α-MSH nor γ-MSH had any effect on androgen production in vitro at any age. Fetal testosterone levels were normal in mice that lack circulating ACTH (proopiomelanocortin-null) indicating that ACTH is not essential for fetal Leydig cell function. Results show that both LH and ACTH can regulate testicular steroidogenesis during fetal development in the mouse and suggest that fetal Leydig cells, but not adult Leydig cells, are sensitive to ACTH stimulation.

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177. ANTIBODIES AGAINST BMPR-IB UNCOVERS A PARACRINE FUNCTION FOR THE RECEPTOR IN MALE MOUSE LEYDIG CELL TESTOSTERONE PRODUCTION IN VITRO

Reproduction Fertility and Development ◽

10.1071/srb10abs177 ◽

2010 ◽

Vol 22 (9) ◽

pp. 95

Author(s):

I. M. Ciller ◽

U. A. Ciller ◽

J. R. McFarlane

Keyword(s):

Cell Culture ◽

Leydig Cell ◽

Leydig Cells ◽

Chorionic Gonadotrophin ◽

Slice Culture ◽

Type I ◽

Paracrine Signalling ◽

Testosterone Production ◽

Bmp Receptors

The male reproductive system is regulated by pituitary gonadotrophins and local testicular factors including the transforming growth factor-β superfamily members which up-regulate and modulate testosterone production respectively. Type I and type II bone morphogenetic protein (BMP) receptors have been identified in both steroidogenic and non-steroidogenic cells in the testis and have been reported to impact steroid production via their effect on steroidogenic enzymes. In this study we investigated if BMPR-IB had an autocrine or paracine role in testosterone production using BMPR-IB antibodies in testis tissue and Leydig cell culture in vitro. Immature (21d) and mature (60d) mice were sacrificed by CO2 asphyxiation and the testis removed, decapsulated and cultured in basal and equine chorionic gonadotrophin (eCG)-conditioned media for three hours at 32 °C in 5 % CO2, in the presence and absence of anti-BMPR-IB. Additionally, Leydig cells were Percoll purified from adult mouse testicular interstitial cells and cultured for three hours with and without anti-BMPR-IB under human chorionic gonadotrophin (hCG) stimulated or unstimulated conditions. After three hours of incubation the culture media was aspirated into labeled vials and assayed for testosterone using a radioimmunoassay. In adult testicular slice culture, treatment with anti-BMPR-IB resulted in a significant decrease in basal and eCG-stimulated testosterone production by 37 % and 41 % respectively, while having no significant effect on basal or eCG-stimulated testosterone production by the immature testis. In purified Leydig cell culture from adult male mice BMPR-IB immunization had no effect on testosterone production in basal or hCG-stimulated conditions. In conclusion, anti-BMPR-IB significantly reduced testosterone production in adult testicular slice culture but not in cell culture, demonstrating that BMP paracrine signalling from the seminiferous tubules is likely to be important in modulating testosterone production by Leydig cells. Additionally, the paracrine signalling appears to be developmentally regulated only occurring in the adult testis.

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Stimulatory and inhibitory factors of Leydig cell steroidogenesis are secreted simultaneously by the rat seminiferous tubules and do not affect Leydig cell inhibin production in vitro

Reproduction Fertility and Development ◽

10.1071/rd9940693 ◽

1994 ◽

Vol 6 (6) ◽

pp. 693 ◽

Cited By ~ 4

Author(s):

JR McFarlane ◽

Kretser DM de ◽

GP Risbridger

Keyword(s):

Conditioned Medium ◽

Leydig Cell ◽

Leydig Cells ◽

Seminiferous Tubules ◽

Low Doses ◽

Adult Rat ◽

Testosterone Production ◽

Significant Reversal ◽

The Mean

The effect of conditioned medium from rat seminiferous tubules (at Stages VII-VIII and Stages IX-VI) cultured with or without follicle-stimulating hormone (FSH) on the production of testosterone and immunoactive inhibin by Leydig cells was examined. Low doses of conditioned medium from unstimulated tubules at Stages VII-VIII significantly (P < 0.05) increased the mean testosterone production to greater than 31 +/- 11% over that achieved with luteinizing hormone (LH) alone. At the highest dose, the conditioned medium significantly inhibited (P < 0.05) LH-stimulated testosterone production by 13 +/- 7%. Low doses of conditioned medium from unstimulated tubules at Stages IX-VI increased the mean testosterone production to 22 +/- 10%, whereas at higher doses, a significant reversal in the stimulation occurred although not to the same extent as that found with medium from tubules at Stages VII-VIII. Conditioned medium from FSH-stimulated tubules at Stages VII-VIII and Stages IX-VI, significantly increased testosterone production to 39 +/- 7% and 31 +/- 13% respectively. Immunoactive inhibin production by the Leydig cells remained unaffected by exposure to conditioned medium from FSH stimulated and unstimulated tubules at Stages VII-VIII and Stages IX-VI. The data demonstrate that tubule culture medium contains FSH-modulated activities which can specifically stimulate and inhibit testosterone synthesis by adult rat Leydig cells in vitro and therefore explains the contradictory reports in the literature.

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Cyclooxygenases in Rat Leydig Cells: Effects of Luteinizing Hormone and Aging

Endocrinology ◽

10.1210/en.2006-0925 ◽

2007 ◽

Vol 148 (2) ◽

pp. 735-742 ◽

Cited By ~ 20

Author(s):

Haolin Chen ◽

Lindi Luo ◽

June Liu ◽

Barry R. Zirkin

Keyword(s):

Leydig Cell ◽

Leydig Cells ◽

Close Correlation ◽

Dibutyryl Camp ◽

Protein Levels ◽

Testosterone Production ◽

Age Related ◽

The Relationship

Previous studies suggested that increased Leydig cell cyclooxygenase (COX)2 expression may be involved in the reduced testosterone production that characterizes aged Leydig cells. Our objective herein was to further elucidate the relationships among LH stimulation, Leydig cell COX2 and COX1 expression, aging, and testosterone production. Incubation of Leydig cells from young or aged rats with LH or dibutyryl cAMP resulted in increases in both intracellular COX2 protein expression and testosterone production. COX1 expression did not respond to LH or dibutyryl cAMP. Incubation of adult cells with a protein kinase A inhibitor suppressed the stimulatory effects of LH on COX2 and testosterone production. Short-term incubation of Leydig cells with TGF-α or IL-1β also increased COX2 protein levels; IGF-I had no effect. In vivo, LH also was found to stimulate both COX2 and testosterone, but not COX1. As reported previously, COX2 expression was greater in old than in young cells, and old Leydig cells responded to inhibition of COX2 in vitro with increased testosterone production. However, the effects of the COX2 inhibitors were not restricted to old cells; young Leydig cells also responded to COX2 inhibition with increased testosterone production. This and the observation that the incubation of young or old cells with LH resulted in increased COX2 and testosterone production in both cases suggests that the relationship between COX2 and testosterone production is not unique to aged Leydig cells. Moreover, the close correlation between increases in COX2 and testosterone in LH-stimulated young and aged Leydig cells is difficult to reconcile with the contention that the increased expression of COX2 in aged cells is responsible for age-related suppression of Leydig cell testosterone production.

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TESTOSTERONE SECRETION BY FOETAL RAT TESTES IN VITRO

Journal of Endocrinology ◽

10.1677/joe.0.0710231 ◽

1976 ◽

Vol 71 (2) ◽

pp. 231-238 ◽

Cited By ~ 67

Author(s):

RÉGINE PICON

Keyword(s):

Cyclic Amp ◽

Synthetic Medium ◽

Functional Differentiation ◽

Dibutyryl Cyclic Amp ◽

Testosterone Secretion ◽

Testosterone Production ◽

Foetal Rat

SUMMARY Testosterone secretion by foetal rat testes (13½–21½ days of gestation) explanted for 3 days in a synthetic medium was measured every 24 h by radioimmunoassay. During the first day of explantation, the foetal testis produced, respectively, 1013 ± 132, 8734 ± 1118, 9179 ± 2185 and 3886 ± 309 (s.e.m.) pg/testis when explanted at 14½, 16½, 18½ and 21½ days respectively. Testosterone production by 13½-day-old testes was not detectable on the first day of culture, but appeared on subsequent days. Daily testosterone secretion increased on the 2nd and 3rd days of culture in 14½-day-old testes and decreased in older stages. These results suggest that the functional differentiation of the testis is independent of stimulatory factors like gonadotrophins. Dibutyryl cyclic AMP was found to stimulate testosterone production significantly from 14½ days of gestation onwards.

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