Stimulated Testosterone Production Research Articles

Effects of Neonatal Exposure to 4-Tert-Octylphenol, Diethylstilbestrol, and Flutamide on Steroidogenesis in Infantile Rat Testis

Exposure of neonatal testis, populated by fetal-type Leydig cells, to endocrine-active compounds may have far-reaching consequences. Our aim was to resolve the sensitivity of testosterone synthesis of infant rat (Sprague-Dawley) testis to diethylstilbestrol (DES; 0.1-1.0 mg/kg), 4-tert-octylphenol (OP; 10-100 mg/kg), and Flutamide (FLU; 2.0-25 mg/kg) given by daily sc injections from birth to postnatal day 4. Testes and serum were collected on day 14 when body and testis weight, testicular histology, circulating testosterone, LH and FSH levels, and steroidogenic acute regulatory protein (StAR) and 3beta-hydroxy-steroid-dehydrogenase (3beta-HSD) protein levels were determined. DES at each dose and FLU at 25 mg/kg dose reduced testis weight and the diameter of seminiferous cords. FLU caused some Leydig cell hyperplasia. Plasma testosterone was reduced in all DES animals, LH elevated in DES 0.5 mg/kg and FLU 25 mg/kg animals, and FSH reduced in the DES 1.0 mg/kg group. Basal testicular ex vivo progesterone and human chorionic gonadotropin (hCG)-stimulated testosterone production were decreased in DES animals. Despite a decrease in hCG-induced cyclic adenosine-3',5'-monophosphate (cAMP) production, intratesticular testosterone was increased in the FLU 10 and 25 mg/kg groups. OP 100 mg/kg elevated hCG-induced progesterone production only. No changes were seen in 3beta-HSD protein levels in any treatment group. StAR levels were reduced in DES animals. The results indicate the sensitivity of postnatal fetal-type Leydig cells to endocrine-active compounds. Suppression of StAR expression level was an early sign of the DES-induced steroidogenic lesion. FLU-induced changes suggest the importance of androgen receptor-mediated regulation of testosterone synthesis in the postnatal rat testis. Octylphenol appeared less effective in bringing about acute steroidogenic changes.

Investigation, Treatment, and Monitoring of Late‐Onset Hypogonadism in Males: ISA, ISSAM, and EAU Recommendations

Demographic data clearly demonstrate that the percentage of the population in the older age group is increasing. Androgen deficiency in the aging male has become a topic of increasing interest and debate throughout the world. Cross-sectional and longitudinal data indicate that the testosterone falls progressively with age and that a significant percentage of men over the age of 60 years have serum testosterone levels that are below the lower limits of young adult (age 20–30 years) men (1–4). The principal questions raised by these observations are whether older hypogonadal men will benefit from testosterone treatment and what will be the risks associated with such intervention. The past decade has brought evidence of benefit of androgen treatment of hypogonadal men on multiple target organs and the recent studies show short-term beneficial effects of testosterone in older men that are similar to those in younger men. This has been comprehensively reviewed and summarized by the Institute of Medicine in ‘Testosterone and Aging: Clinical Research Directions’ (5). Long-term data on the effects of testosterone treatment in the older population are limited mainly to effects on body composition and bone mass (6–11). Key questions of the effects of testosterone on patient reported outcomes and functional benefits that may retard physical or mental frailty of the elderly or improve the quality of life are not yet available. Specific risk data on the prostate and cardiovascular systems are needed.

Hypothalamic and Gonadal Components of Hypogonadism in Boys with Prader-Labhart- Willi Syndrome

The specific form of hypogonadism in Prader-Labhart-Willi syndrome (PWS), central or peripheral, remains unexplained. The objectives of this study were to investigate the cause of hypogonadism in PWS and determine whether human chorionic gonadotropin (hCG) treatment can restore pubertal development. This was a clinical follow-up study, divided into two samples, over a duration of 1.5 and 4.5 yr. Eight male infants and six peripubertal boys (age at start of observation, 0.06-0.93 and 8.1-10.8 yr, respectively) with genetically confirmed PWS were studied. hCG (500-1500 U twice weekly) was given from age 13.5 yr to the present. Serum FSH, LH, inhibin B, and testosterone levels and pubertal development were the main outcome measures. Infants with PWS presented normal LH (2.3 +/- 0.7 U/liter) and testosterone (2.5 +/- 0.9 nmol/liter) levels (mean +/- sem at 5 months) compared with the reference range. However, two thirds of the boys displayed cryptorchidism. Inhibin B levels were at the lowest level of the normal range and decreased significantly between infancy and puberty (at 13 yr, 72 +/- 17 pg/ml), whereas FSH secretion increased (9.9 +/- 2.6 U/liter). Pubertal maturation stopped at an average bone age of 13.9 yr. hCG therapy increased testosterone (11 +/- 2 nmol/liter) and reduced FSH (at 16 yr, 1.1 +/- 0.9 U/liter) levels. Testicular volume (5.6 +/- 1 ml) and inhibin B (26.5 +/- 11.9 pg/ml) remained low. Children with PWS display a specific form of combined hypothalamic (low LH) and peripheral (low inhibin B and high FSH) hypogonadism, suggesting a primary defect in Sertoli and/or germ cell maturation or an early germ cell loss. hCG therapy stimulates testosterone production and virilization.

Hormonally Active Nontransformed Human Ovarian Cell Culture from Oophorectomy Specimens: Methods of Development and Initial Characterization

We repeatedly established a nontransformed steroidogenically active human ovarian cell culture derived from oophorectomy specimens. The cells maintained steroidogenic activity for 3–5 passages (6–8 weeks) and responded to stimulation by insulin and gonadotropin. With pregnenolone as substrate, LH stimulated progesterone production up to 124% and FSH up to 121%. Insulin alone stimulated progesterone production up to 135%, in the presence of LH up to 191%, and in the presence of FSH up to 170%. With dehydroisoandrosterone (DHA) as substrate, insulin alone stimulated testosterone production up to 117%, and in the presence of LH (but not FSH) up to 125%. With androstenedione as substrate, insulin alone stimulated estradiol production up to 133%, FSH alone up to 188%, and LH with insulin up to 217%. With progesterone as substrate and in the presence of LH (but not FSH), 17-α-hydroxylase activity was stimulated up to 131%. With DHA as substrate and in the presence of LH, 3-β-hydroxysteroid dehydrogenase (3-β-HSD) activity was stimulated up to 139%. With androstenedione as substrate, insulin alone stimulated aromatase activity up to 202%, LH up to 208%, and FSH up to 251%. Under the same conditions, in the presence of LH and insulin, aromatase activity was stimulated up to 342%, and in the presence of FSH and insulin, up to 318%. With testosterone as substrate, insulin alone stimulated aromatase activity up to 122%. With testosterone as substrate, in the presence of LH and insulin, aromatase activity was stimulated up to 136%, and in the presence of FSH and insulin, up to 156%. Immunocytochemistry studies directly confirmed presence of aromatase and 3-β-HSD in these cultured cells. We conclude that a steroidogenically active nontransformed long-term human ovarian cell culture can be repeatedly established from oophorectomy specimens, providing uninterrupted supply of cultured human ovarian cells for a variety of studies of ovarian physiology.

Stimulation of testosterone production in rat Leydig cells by aldosterone is mineralocorticoid receptor mediated

The testis is known to be a site of corticosterone action, and testosterone production in Leydig cells is directly inhibited by glucocorticoids. Glucocorticoids bind to both glucocorticoid receptors (GRs) and to mineralocorticoid receptors (MRs). In Leydig cells, selective mineralocorticoid binding could result from oxidative inactivation of glucocorticoid by type 1 and/or 2 11β-hydroxysteroid dehydrogenase (11βHSD), as both isoforms are expressed. However, it remains unclear whether Leydig cells express MRs and respond directly to mineralocorticoid action. Therefore, the aims of the present study were to ascertain: (1) whether MR mRNA, protein and receptor binding are present in Leydig cells; and (2) if the mineralocorticoid modulates testosterone production. The mRNA encoding MR, as well as protein, and binding activity were each observed in adult rat Leydig cells. MR-ligand binding specificity within isolated Leydig cells was evaluated further by measuring displacement of MR binding to aldosterone by corticosterone in the presence and absence of carbenoxolone, an inhibitor of 11βHSD1 and 2 that decreases conversion to biologically inert 11-dehydrocorticosterone. Carbenoxolone inhibited 11βHSD oxidative activity, and reduced corticosterone-binding by 50%. Mineralocorticoid effects on steroidogenesis were assessed in the presence of aldosterone (0.01–10 nM) with or without the MR antagonist, RU28318. Aldosterone induced dose-dependent increases in both basal and luteinizing hormone-stimulated testosterone production. RU28318 eliminated the increase, indicating that these effects of aldosterone were mediated by the MR. The effects of aldosterone and luteinizing hormone (0.1 ng/ml) on testosterone production were synergistic, suggesting that the two hormones increased steroidogenesis through separate pathways. We conclude that Leydig cells express MRs and that testosterone production is subject to regulation by aldosterone.

Regulatory mechanism of Toona sinensis on mouse leydig cell steroidogenesis

Toona sinensis (TS), a kind of arbor, widely distributes nowadays in Asia. The leaves of TS have been used as an effective nutritious food in Chinese society for a long time. It was reported that Toona sinensis can induce apoptosis of cancer cells, reduce plasma glucose in diabetic rats, and improve lipolysis of differentiated 3T3-L1 adipocyte and its uptake of glucose. It has also been shown that TS may increase dynamic activity of human sperm. Thus, we are interested to investigate whether Toona sinensis has any effect on mouse Leydig cell testosterone production, which correlates to sperm activity. Primary mouse Leydig cells were purified to conduct the in vitro experiments. Different concentrations of crude Toona sinensis were added to primary mouse Leydig cells and the testosterone production was determined. The results showed that crude TS significantly inhibited both basal and human chorionic gonadotropin (hCG)-stimulated testosterone productions in dose dependent manner, respectively ( P<0.05). Crude TS also reduced the forskolin- and dibutyryl-cAMP (dbcAMP)-stimulated testosterone production ( P<0.05), which indicated that crude TS might affect protein kinase A (PKA) signal transduction pathway at the site after the formation of cyclic AMP. Moreover, TS inhibited Leydig cell steroidogenesis by suppressing the activity of steroidogenic enzymes including P450 side chain cleavage enzyme, 3β-hydroxysteroid dehydrogenase, 17α-hydroxylase, 20α-hydroxylase and 17β-hydroxysteroid dehydrogenase ( P<0.05). In summary, these results suggested that TS inhibited steroidogenesis by suppressing the cAMP-PKA signaling pathway and the activities of steroidogenic enzymes in normal mouse Leydig cells.

Fish eggs as bioreactors: the production of bioactive luteinizing hormone in transgenic trout embryos

We demonstrated the production of goldfish luteinizing hormone (gfLH) by the use of 4-day-old rainbow trout embryos as novel bioreactors. This expression system has several advantages: target proteins can be rapidly expressed at low cost, and recombinant proteins can be synthesized at low temperatures and can undergo complex post-translational modifications (PTMs). An expression vector containing gfLH cDNA was microinjected into fertilized trout eggs. After 4 days of incubation at 10 degrees C, transgenic embryos were harvested and glycosylated recombinant gfLH was recovered, which stimulated testosterone production in testicular fragments from the goldfish. This is the first report on the successful production of bioactive recombinant gonadotropin originated from cyprinid. Further, these results demonstrate that trout-embryo bioreactors are a potentially powerful tool for the production of functional recombinant proteins.

Endocrine and paracrine control of Leydig cell steroidogenesis and proliferation in the wall lizard: an in vitro study

The present in vitro study, for the first time, demonstrates the endocrine as well as paracrine control of Leydig cell steroidogenesis and proliferation in the wall lizard Hemidactylus flaviviridis. Unlike mammals, Leydig cell activity in the wall lizard seems to be directly controlled by ovine follicle-stimulating hormone (FSH)-like molecule, since FSH increased the testosterone production and tritiated thymidine ([ 3H]TdR) incorporation by Leydig cells. In addition, Sertoli cell paracrine factor or factors play important roles in controlling Leydig cell activities as non-activated Sertoli cell-conditioned medium (SCCM) alone stimulated testosterone production by both non-activated and FSH-preactivated Leydig cells. As far as the proliferation was concerned, non-activated SCCM did not influence [ 3H]TdR uptake by non-activated or FSH-preactivated Leydig cells, while FSH-preactivated SCCM was able to stimulate proliferation of activated Leydig cells. It may be concluded that FSH, besides directly controlling, also regulates Leydig cell activities indirectly through stimulating the secretion of Sertoli cell paracrine factors. Moreover, steroidogenic factor is different from mitogenic factor because non-activated Leydig cells were responsive to steroidogenic factor but nonresponsive to mitogenic factor.

Effects of thyroid hormones on Leydig cells in the postnatal testis.

Thyroid hormones (TH) stimulate oxidative metabolism in many tissues in the body, but testis is not one of them. Therefore, in this sense, testis is not considered as a target organ for TH. However, recent findings clearly show that TH have significant functions on the testis in general, and Leydig cells in particular; this begins from the onset of their differentiation through aging. Some of these functions include triggering the Leydig stem cells to differentiate, producing increased numbers of Leydig cells during differentiation by causing proliferation of Leydig stem cells and progenitors, stimulation of the Leydig cell steroidogenic function and cellular maintenance. The mechanism of action of TH on Leydig cell differentiation is still not clear and needs to be determined in future studies. However, some information on the mechanisms of TH action on Leydig cell steroidogenesis is available. TH acutely stimulate testosterone production by the Leydig cells in vitro via stimulating the production of steroidogenic acute regulatory protein (StAR) and StAR mRNA in Leydig cells; StAR is associated with intracellular trafficking of cholesterol into the mitochondria during steroid hormone synthesis. However, the presence and/or the types of TH receptors in Leydig cells and other cell types of the Leydig cell lineage is still to be resolved. Additionally, it has been shown that thyrotropin-releasing hormone (TRH), TRH receptor and TRH mRNA in the testis in many mammalian species are seen exclusively in Leydig cells. Although the significance of the latter observations are yet to be determined, these findings prompt whether hypothalamo-pituitary-thyroid axis and hypothalamo-pituitary-testis axis are short-looped through Leydig cells.

Effect of bullfrog LH and FSH on newt testes under different temperatures

Environmental temperature plays important roles for amphibian gonadal function. In this study, we examined the responses of testicular tissue of adult male newts ( Cynops pyrrhogaster) to amphibian gonadotropins both in vitro and in vivo under different temperatures. When minced testes were incubated in vitro at different temperatures (8–37 °C) under an atmosphere of 95% O 2–5% CO 2 for 3 h with bullfrog luteinizing hormone (LH) or follicle-stimulating hormone (FSH), LH stimulated testosterone production more than FSH. The testosterone production increased as the incubation temperature increased. Hypophysectomized newts were injected with bullfrog LH or FSH and maintained at 8 or 18 °C. In the 18 °C group, the testicular weight of the hypophysectomized control decreased when compared with that of with the intact control. The testicular weight of the LH-treated hypophysectomized group decreased more than that of the hypophysectomized control, indicating that LH induced the evacuation of mature spermatozoa from the testes of LH-treated hypophysectomized newts. In the FSH-treated newts, the testicular weight was greater than that in the hypophysectomized control, and was maintained at a value similar to that of the intact control. In the 8 °C group, there was no significant difference in testicular size among the intact control, hypophysectomized control, and FSH-treated newts. LH strongly induced spermiation as it did at 18 °C. The plasma testosterone level in the hypophysectomized newts decreased dramatically, but LH was effective in restoring it. Its effect was more potent at 8 °C than at 18 °C. On the other hand, FSH did not induce a significant increase in the plasma testosterone levels at either temperatures. The results indicate a temperature-dependent difference in responsiveness of the testis both in vitro and in vivo to LH and FSH.

Effects of Aqueous Extracts of "Betel Quid" and Its Constituents on Testosterone Production by Dispersed Mouse Interstitial Cells

Betel quid (BQ) is a favorite chewing item among many communities in different parts of Asia where it is popular by different names. BQ is a unique combination of nut or fruit from the Areca catechu Linn. (AN) tree, leaf from the Piper betle Linn. (BL) vine, slaked lime, paste of bark from the Acacia catechu tree and other spices. AN has been used successfully in various traditional medicines by different civilizations over several ages. Initially condemned by the medical communities for its health hazards, identification and application of potent pharmacologically bioactive compounds from different constituents of BQ have rekindled growing interest in related investigations. Curious about the stimulating role of BQ, we investigated the potential steroidogenic activity of hot water extract from BQ and its constituents and arecoline on testosterone producing ability in an in vitro experiment. Enzyme dissociated interstitial cells from adult mouse testes (ICR strain) were cultured with/without different doses of the extracts and the level of testosterone produced was assayed by an enzyme immunoassay (EIA) technique. It was found that at lower doses of arecoline, AN and BL extracts had significantly stimulated testosterone production over the basal level (p < 0.05). BQ extract, on the other hand, did not show any significant effect on testosterone production. Combinations of arecoline at low doses with 10 ng/ml ovine leutinizing hormone (oLH) showed increases in testosterone produced, while cyclic adenosine monophosphate (cAMP) co-culture showed dose-related inhibition. Our current finding hints at the possible dose-dependent dualistic role of AN and BL extracts and arecoline for testosterone production employing possible non-cAMP-dependent pathway of steroidogenesis. However, the identity of the active compounds besides arecoline and the exact mechanism involved remains to be further investigated.

Adrenocorticotropic hormone directly stimulates testosterone production by the fetal and neonatal mouse testis.

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 ED(50) for ACTH, measured in isolated fetal and neonatal testicular cells, was 5 x 10(-10) M and the lowest dose of ACTH eliciting a response was 2 x 10(-11) M. Circulating ACTH levels in fetal mice were around 8 x 10(-11) M. Neither alpha-MSH nor gamma-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.

Time- and dose-related effects of estradiol and diethylstilbestrol on the morphology and function of the fetal rat testis in culture.

The mechanisms underlying the action of estrogens in the fetal testis are still largely obscure. In particular, whether this action is direct or indirect remains largely unexplored. This study was aimed at investigating the effect of estradiol (E2) and diethylstilbestrol (DES) on the testis from 14.5-day-old rat fetuses in culture, at concentrations ranging from 4 x 10(-10) M (Kd of E2 for the estrogen receptors [ER]: 1-4 x 10(-10) M) to 4 x 10(-6) M (concentration previously shown in the literature to affect in vitro gonocyte proliferation). Exposure to DES and E2 decreased gonocyte number, the effects of DES being much more drastic than those of E2. Gonocyte number decreased in a concentration-dependent manner (day 3: -5%, -16%, and -80% at 4 x 10(-10) M, 4 x 10(-8) M, and 4 x 10(-6)M of DES, respectively), as well as in a time-dependent manner (at 4 x 10(-6) M DES: -31% on day 1, -60% on day 2, and -80% on day 3). This was due to a decrease in the gonocyte mitotic index and a dramatic increase in apoptosis. Importantly, in the presence of the anti-estrogen ICI 182.780 (ICI), the effect of DES was abolished. Sertoli cell number subsequently decreased (day 3), although the rate of apoptosis did not increase. These changes were less dramatic than those observed with gonocytes and were due to a decrease in Sertoli cell proliferation, which was not antagonized by ICI. Addition of 4 x 10(-6) M DES had no effect on basal Sertoli cell cyclic adenosine 5'-monophosphate (cAMP) levels or on follicle-stimulating hormone (FSH)-stimulated cAMP production after adjustment for Sertoli cell number. Finally, estrogens reduced both Leydig cell number (-26% on day 3, 4 x 10(-6) M DES) and basal and luteinizing hormone (LH)-stimulated testosterone production. The latter effects were antagonized by ICI. 1) E2 and DES induce alterations in the germ line and in somatic cells; 2) gonocyte alteration was the first event detected, and the action of estrogens at this level was mediated by ER, as is the case in Leydig cells; and 3) these data should help us to understand estrogen effects on the fetus and should be considered in the context of the debate on environmental estrogens.

Clinical review 148: Monogenic disorders of puberty.

Normal puberty is associated with the onset and progressive activation of the hypothalamic-pituitary-gonadal (HPG) axis and the resultant development of secondary sexual characteristics. Puberty begins with increasing nocturnal pulsatile hypothalamic GnRH secretion, which gradually occurs throughout the 24-h day. Pulsatile GnRH stimulates pituitary FSH and LH secretion, ultimately stimulating gonadal steroid production and gametogenesis in females and males. In the ovary, FSH stimulates follicular maturation and estrogen production through aromatization of androgens, whereas LH stimulates androgen production by theca cells, triggers ovulation, and maintains progesterone production by the corpus luteum. In the testis, FSH acts on Sertoli cells to initiate spermatogenesis and LH acts on Leydig cells to stimulate testosterone production. The hypothalamic-pituitary-adrenal (HPA) axis also has some minor input into the physiological process of puberty, through the secretion of adrenal androgens. However, the major involvement of the HPA axis in puberty is in its potential pathological influence, primarily in accelerating its onset and/or progress. Traditionally, the onset of adrenal androgen secretion in childhood has been termed “adrenarche,” whereas the onset of gonadal steroid secretion has been called “gonadarche.” Delayed puberty describes the clinical condition in which the pubertal events start late or are attenuated or arrested. In contrast, precocious puberty describes the clinical condition in which the pubertal events start early. Mutations have been identified in an increasing number of genes that influence the onset and progression of puberty. These discoveries have provided new insights into the physiology and pathophysiology of this important life transition and have greatly influenced the practice of reproductive medicine. For instance, the concept of two gonadotropins acting on two separate cell types in the gonad has been the cornerstone of reproductive endocrinology, while it has been traditionally believed that both FSH and LH are required for fertility in females and males. However, studies of mutant gonadotropin receptors indicate that female reproductive capacity depends primarily on FSH, whereas male reproductive capacity depends primarily on LH. We summarize the molecular defects that influence gonadal and/or adrenal function and cause delayed (Table 1) or precocious puberty (Table 2). Here, we present genes whose alterations result in abnormal puberty in girls and boys.

Breaking Up the T Party: Short-circuited hormone signal keeps old rats from making testosterone (Sex hormones)

Women haven't cornered the market on hormonal woes. As men age, their blood testosterone concentrations decline, which might contribute to health problems such as osteoporosis (see "More Than a Hot Flash" ). A new study casts blame on a withered signaling pathway that normally jolts testosterone-producing cells into action. Scientists don't understand what triggers the fall in testosterone concentration, but the Brown Norway rat, which experiences an ebb in testosterone concentration similar to that in humans, has provided some clues. In old animals, testosterone factories in the testes called Leydig cells lose their capacity to churn out the hormone. Previous studies indicated that supplemental luteinizing hormone (LH), which normally prods Leydig cells into action, couldn't elevate testosterone concentrations in old rats. To find out why, Chen and colleagues removed Leydig cells from young and old rats and cultured them in the presence of LH. Cells from the youngsters pumped out testosterone, but those from geezer rodents did not. To investigate why old cells fail to respond to LH, the team measured how much LH bound to each type of cell. Old cells glommed onto one-third fewer hormone molecules than young cells did, indicating a decrease in the number of cell surface receptors that bind LH. But a lack of receptors didn't cause the decline in testosterone production, according to the outcome of an additional experiment. The researchers collected Leydig cells from young animals whose LH output had been artificially lowered. These cells had fewer LH receptors--yet they made much more testosterone in culture than old cells did, a result that takes the number of LH receptors off the hook. Perhaps the pathway through which the LH receptor stimulates testosterone production is defective, the researchers proposed. After binding LH, the receptor triggers synthesis of a molecule called cyclic AMP (cAMP) inside Leydig cells; cAMP ignites testosterone production. Chen and colleagues found that cells from old rats contained lower cAMP concentrations than did those from either normal young animals or young animals with reduced LH. A general slump in cAMP formation wasn't to blame: Old cells pumped out the molecule when treated with a chemical that directly stimulates the cAMP-producing enzyme. The results suggest that aging short-circuits the LH receptor's ability to trigger cAMP synthesis in Leydig cells. How this glitch arises is mysterious, but the authors propose that Leydig cells might accrue oxidative damage while making testosterone. Enzymes generate reactive oxygen species as they manufacture testosterone, and previous work by the team suggests that decreasing testosterone output reduces oxidative damage in Leydig cells. The study provides new evidence for how rat Leydig cells lose their capacity to generate testosterone, says endocrinologist Alvin Matsumoto of the University of Washington, Seattle, but it's uncertain whether humans suffer from the same defect. The idea that free radicals assault Leydig cells is an intriguing one, although not proven, he says. If that's true, antioxidants might help maintain testosterone production. Perhaps the delicate broccoli flower will be the secret to youthful vigor in older men. --R. John Davenport; suggested by James M. Harper H. Chen, M. P. Hardy, B. R. Zirkin, Age-related decreases in Leydig cell testosterone production are not restored by exposure to LH in vitro. Endocrinology 143 , 1637-1642 (2002). [Abstract] [Full Text]

Endocrine regulation of testicular function in men: implications for contraceptive development

The testicle has two important and complimentary functions in the adult male: the production of sperm and the synthesis of testosterone. Testicular function is directed by the central nervous system through its effects on the pituitary gland. Precise regulation of testicular function is accomplished by an elegant endocrine feedback loop in which the secretion of pituitary gonadotropins is stimulated by the pulsatile release of GnRH from the hypothalamus and modulated by testicular hormones. Inhibin, testosterone and its metabolites inhibit the secretion of the gonadotropins both directly at the pituitary and centrally at the level of the hypothalamus. In the testis, LH mainly stimulates testosterone production and FSH promotes spermatogenesis, but the exact details of these actions are only now being elucidated. The discovery of novel mutations in gonadotropin structure and signaling provide unique insight into the roles of these crucial hormones in the development and function of the male reproductive axis. A key goal of research into hormonal regulation of testicular function is the development of reversible, safe and effective male hormonal contraceptives. Recent promising trials of hormonal contraceptive combinations may soon bring the promise of male contraception to fruition.

Metyrapone-induced corticosterone deficiency impairs glucose oxidation and steroidogenesis in Leydig cells of adult albino rats.

The present study was designed to identify the effects of metyrapone-induced corticosterone deficiency on Leydig cell steroidogenesis in adult male rats. Adult Wistar rats (200-250 g body weight) were treated with metyrapone, an inhibitor of corticosterone synthesis (10 mg/100 g body weight, s.c., twice daily) for 10 days. Experimental animals were killed along with controls, blood was collected, and sera separated for testosterone and estradiol assays. Testes were removed and Leydig cells were isolated, purified and used for estimating the specific activity of 17beta-hydroxysteroid dehydrogenase (17beta-HSD) and 14C-glucose oxidation. Serum testosterone (p < 0.05), Leydig cellular 14C-glucose oxidation (p < 0.001) and the specific activity of 17beta-HSD (p < 0.01) were significantly decreased in metyrapone treated rats. However, serum estradiol was not markedly altered compared to control. In addition to this, a set of in vitro experiments were also performed to identify the effects of metyrapone-induced corticosterone deficiency on hCG and prolactin-induced Leydig cell testosterone production. Metyrapone treatment significantly (p < 0.05) decreased the Leydig cellular basal as well as hCG and its combination with prolactin stimulated testosterone production in vitro. It is concluded from the present study that the inhibitory effects of metyrapone-induced corticosterone deficiency on Leydig cell steroidogenesis are mediated through impaired glucose oxidation and 17beta-HSD activity. In vitro studies showed that corticosterone deficiency impairs not only hCG action but also the potentiating effect of prolactin on Leydig cell steroidogenesis.

Effects of Cordyceps sinensis on testosterone production in normal mouse Leydig cells

The stimulatory effect of Cordyceps sinensis (CS) on MA-10 mouse Leydig tumor cell steroidogenesis was previously demonstrated in our laboratory. In the present studies, we further determined the effect of CS on steroidogenesis in purified normal mouse Leydig cells. Different concentrations of CS (0.1–10 mg/ml) were added to Leydig cells without or with human chorionic gonadotropin (hCG) (50 ng/ml), and the steroid production was determined by radioimmunoassay (RIA). The results illustrated that CS stimulated normal mouse Leydig cell steroidogenesis in a dose-dependent relationship. CS at 3 mg/ml significantly stimulated testosterone production (p<0.05). Concerning the temporal relationship, CS at 3 mg/ml stimulated maximal testosterone production between 2 to 3 hr. Interestingly, hCG-stimulated testosterone productions were suppressed by CS in a dose-dependent relationship. CS also reduced dbcAMP-stimulated testosterone productions, which indicated that CS affected signal transduction pathway of steroidogenesis after the formation of cyclic AMP. Moreover, cycloheximide inhibited CS-treated mouse Leydig cell testosterone production, suggesting that new protein synthesis was required for CS-stimulated steroidogenesis.

Age-dependent stimulation of Leydig cell steroidogenesis by interleukin-1 isoforms

Different isoforms of testicular interleukin-1 (IL-1) were analysed to determine whether there were differences in the ability to modulate rat Leydig cell steroidogenesis in vitro. Rat 17K IL-1α and IL-1β, 32K IL-1α precursor (32proIL-1α) and a 24K splice variant (24proIL-1α) stimulated testosterone production by Leydig cells from 40- but not 80-day-old rats. The potency of the isoforms was IL-1α>IL-1β>32proIL-1α>24proIL-1α, IL-1α being 50-fold more potent than IL-1β. IL-1 receptor antagonist reversed the effects and IL-1 receptor type I mRNA was expressed by the responding Leydig cells, indicating a receptor mediated action. Inhibition of PKA and Ca 2+ channels abolished IL-1-induced steroidogenesis, while inhibition of PKC had no significant effect. Except for 24proIL-1α which was stimulatory, all IL-1 isoforms suppressed hCG-driven testosterone production. This inhibitory effect was abolished by androstendione, suggesting that P450c17 was suppressed by IL-1. Our results indicate that IL-1 plays a paracrine role in the regulation of Leydig cell steroidogenesis.

Endocrine regulation of testicular function in men: implications for contraceptive development

The testicle has two important and complimentary functions in the adult male: the production of sperm and the synthesis of testosterone. Testicular function is directed by the central nervous system through its effects on the pituitary gland. Precise regulation of testicular function is accomplished by an elegant endocrine feedback loop in which the secretion of pituitary gonadotropins is stimulated by the pulsatile release of GnRH from the hypothalamus and modulated by testicular hormones. Inhibin, testosterone and its metabolites inhibit the secretion of the gonadotropins both directly at the pituitary and centrally at the level of the hypothalamus. In the testis, LH mainly stimulates testosterone production and FSH promotes spermatogenesis, but the exact details of these actions are only now being elucidated. The discovery of novel mutations in gonadotropin structure and signaling provide unique insight into the roles of these crucial hormones in the development and function of the male reproductive axis. A key goal of research into hormonal regulation of testicular function is the development of reversible, safe and effective male hormonal contraceptives. Recent promising trials of hormonal contraceptive combinations may soon bring the promise of male contraception to fruition.