Differential effects of follicle-stimulating and luteinizing hormones on testosterone production by mouse testes

In adult mice, direct intratesticular injection of ovine follicle-stimulating hormone (o-FSH-13; AFP 2846-C, from NIAMDD, less than 1% LH contamination) at 10, 100 or 1000 ng significantly elevated concentrations of testosterone (T) within the testis. These effects were rapid, with peak values attained by 15 min, and transient, with return to values comparable to that in the contralateral, saline-injected testis within 90 min. Intratesticular injection of FSH (1 microgram) significantly increased testicular T levels in 15- and 60-day old mice. This contrasted with the effects of intratesticular administration of human chorionic gonadotropin (hCG), which stimulated T production significantly at 30 days of age through adulthood. In adult mice, the equivalent LH to the possible contamination in the FSH preparation (1 ng) had no effect. Intratesticular injection of 10 ng LH produced comparable stimulation to that by 100 ng FSH (approximately 7-fold). Systemic pre-treatment with a charcoal-treated porcine follicular fluid (PFF) extract for 2 days reduced plasma FSH levels [86 +/- 17 (5) vs 700 +/- 8 (6); P less than 0.05], but had no effect on plasma LH. Twenty-four hours after the last treatment, the response to intratesticular injection of hCG (2.5 mIU), FSH (100 ng) or LH (10 ng) was also significantly attenuated in these mice. Intratesticular injection of PFF had no direct effect on testicular T levels. In vitro T production in the presence of hCG, LH or FSH were differentially affected by the concentrations of calcium (Ca2+) or magnesium (Mg2+) in the incubation media. The stimulatory effects of FSH were apparent at significantly lower levels of Ca2+ or Mg2+, than were those of LH or hCG. The results of these studies indicate that FSH is capable of stimulating testicular T production. Furthermore, the responsiveness to FSH is qualitatively different than that to LH/hCG in terms of the age pattern, as well as the dependence on Ca2+ or Mg2+. In addition, plasma FSH levels appear to influence testicular responsiveness to direct exogenous administration of gonadotropins. These studies indicate that FSH stimulation of T production can be differentiated from those of LH, and that these effects of FSH can be observed under physiological conditions.     

The influence of short photoperiod on testicular and circulating levels of testosterone precursors in the adult golden hamster

The in vivo effects of short photoperiod (SPP, 6L:18D) for 8 and 12 wk on plasma and testicular levels of testosterone (T) precursors in adult golden hamsters were evaluated. Plasma and testicular progesterone (P), 17 alpha-hydroxyprogesterone (17 alpha-OHP), androstenedione (A-dione), and T were measured after 5 injections of saline or human chorionic gonadotropin (hCG) (5 or 25 IU/day). The basal levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and prolactin (PRL) in circulation were also determined. There were significant reductions in the weight of the testes in animals exposed to SPP. After 12 wk in SPP, circulating levels and testicular content of 17 alpha-OHP, A-dione, and T were significantly reduced, suggesting that the decrease in T secretion may be associated with the impairment of synthesis and/or action of 17 alpha-steroid hydroxylase, C17-20 steroid lyase, and 17 beta-hydroxysteroid dehydrogenase enzymes in the testes. Exposure to SPP for 8 wk resulted in decreased plasma and testicular content of T. Although there were reductions in testicular content of 17 alpha-OHP and A-dione, this was not reflected in plasma levels of these steroids. After 8 and 12 wk of exposure to SPP, hCG treatment increased the total amounts of T precursors (except P at 8 wk) in the testes, but the values attained in animals exposed to 12 wk of SPP remained below those observed in hamsters kept in a long photoperiod (14L:10D), suggesting that gonadotropin replacement alone may be insufficient to normalize testicular steroidogenesis.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of short photoperiod on the ability of golden hamster pituitaries to secrete prolactin and gonadotropins in vitro

Transfer of male golden (Syrian) hamsters from a 14L:10D (light:dark) to a 5L:19D photoperiod induced significant changes in pituitary function tested in vitro. Within 27 days after transfer to a 5L:19D photoperiod, basal prolactin (Prl) release was significantly depressed and response to dopamine (DA) was significantly enhanced as compared to Prl release by pituitaries from 14L: 10D hamsters. Follicle-stimulating hormone (FSH) release tended to be depressed after 9 or 27 days of 5L:19D exposure, but the effect was not significant. After 77 days of 5L:19D exposure, Prl release was further suppressed, while FSH release surpassed that seen in 14L:10D pituitaries. In vitro FSH response to luteinizing hormone releasing hormone (LHRH) was also enhanced at this time. After 15 weeks of exposure to a short photoperiod, FSH secretion was still elevated above control levels, but Prl release and Prl response to DA were no longer different from that of 14L: 10D controls. Secretion of luteinizing hormone (LH) in vitro, either basal or LHRH stimulated, was not affected by photoperiod at any time tested. From these results, we conclude that short photoperiod exposure does not reduce the pituitary's ability to secrete LH or FSH, although secretion of Prl is severely attenuated.     

Suppression of sympathetic nervous system by short photoperiod and melatonin in the Syrian hamster

Exposure to short photoperiod or melatonin treatment brings about gonadal regression in Syrian hamsters. The possible influence of these treatments on the sympathetic nervous system (SNS) in these animals was investigated. Male Syrian hamsters were exposed to either long or short photoperiod or subjected to administration of melatonin or its vehicle solution. Exposure of hamsters to 10 weeks of short photoperiod, significantly reduced the noradrenaline (NA) turnover in the heart. Daily administration of melatonin for 8 weeks also resulted in a similar suppression of NA turnover in the heart. Hamsters that were treated with melatonin maintained a lowered metabolic rate as well, at and below thermoneutral temperature. These findings suggest that in a deep hibernator, short photoperiod could suppress the peripheral sympathetic activity and that melatonin may act as the endogenous mediator.     

Effects of photoperiod, beta-endorphin, and naloxone on in vitro secretion of testosterone in white-footed mouse (Peromyscus leucopus) testes

The effects of the pro-opiomelanocortin-derived beta-endorphin (B-EP) and the opioid antagonist naloxone on in vitro secretion (accumulation of testosterone (T) in the medium) of T by testicular cells were assessed in adult white-footed mice (Peromyscus leucopus). Animals were housed under long days (16L:8D) to maintain testicular function or under short days (8L:16D) to induce gonadal regression. In vitro treatment with B-EP or naloxone did not affect basal secretion of T in dispersed cells from active or regressed testes. However, B-EP caused a dose-dependent reduction in secretion of T from cells stimulated maximally with human chorionic gonadotropin (hCG) or dibutyryl cyclic adenosine 3', 5'-monophosphate (dbcAMP). Conversely, naloxone enhanced maximal hCG- and dbcAMP-stimulated secretion of T in testicular incubates from both long- (1.5-fold) and short-day (3.5-fold)-exposed mice. The finding that the addition of naloxone to maximally stimulated cells increased further the secretion of T is evidence that B-EP may act to inhibit gonadotropin-stimulated secretion of T. Also, the stimulatory effect of naloxone on cells from regressed testes indicates that B-EP may be involved in suppressing production of T during the gonadally regressed state. Testicular B-EP-like immunostaining is present within the cytoplasm of interstitial cells and is not apparent in the seminiferous tubules. Together, these results support the idea that in P. leucopus endogenous opioid peptides in the testes may aid in the regulation of testicular function throughout the yearly breeding cycle.     

Modulation of leptin sensitivity by short photoperiod acclimation in the Djungarian hamster, Phodopus sungorus

During seasonal acclimation, Djungarian hamsters spontaneously exhibit a reduction in food intake, body mass and body fat stores, which is externally cued by shortening of day length in autumn and controlled by a sliding set-point. We investigated the function of the leptin adipostatic feedback system in the photoperiodic control of seasonal acclimation. In response to mouse recombinant leptin injections for 10 days, long day photoperiod (LD) and short day photoperiod (SD)-acclimated hamsters decreased food intake and body mass. The reduction of body mass was due to the depletion of body fat, as revealed by carcass composition analysis. In SD hamsters, leptin caused a larger reduction of body fat mass than observed under LD conditions, whereas the anorectic effect was similar in both photoperiods. The serum leptin concentration was 9.3 ± 1.2 ng/ml in LD-acclimated hamsters and decreased significantly to 4.2 ± 0.8 ng/ml and 2.1 ± 0.6 ng/ml in hamsters exposed to SD for 66 days and 116 days, respectively (P < 0.001). A strong positive correlation between total body fat mass and serum leptin concentration was found (r _S=0.935, P < 0.0001, n=70). Despite the anorectic action of exogenous leptin, higher endogenous leptin levels in LD hamsters were paralleled by higher food intake in LD hamsters as compared to SD hamsters. This paradoxical finding further supports the increased leptin sensitivity in SD hamsters as judged from leptin treatment experiments. We tested the functional significance of leptin for the controlled down-regulation of food intake and body mass induced by short photoperiod. Food restriction for 10 days during the transition phase decreased body mass below the desired sliding set-point, which was recovered in control hamsters following ad libitum refeeding. Treatment with mouse recombinant leptin during ad libitum refeeding inhibited the recovery of body mass and blunted the increase of food intake observed in controls, indicating that the sliding set-point utilizes leptin as a signal for the adjustment of the appropriate body mass level. Accepted: 15 October 1999     

Effects of short photoperiod on ATPase activities in the testis of the immature Siberian hamster.

The effects of changes in photoperiod length upon body weight; spleen, thymus, and testis weights; testis protein content; testis cation pump enzyme activities; and plasma testosterone were studied in the developing Siberian hamster, Phodopus sungorus. Male hamsters were exposed to a cycle of 16L:8D (long-day), until Day 18 when half were switched to a 10L:14D (short-day) cycle, until killed 0, 2, 4, 7, 10, 12, or 15 days later. Body weight and relative testis weight (expressed as percentage of body weight) increased steadily during the first week of exposure. After 10 days, the long-day hamsters consistently weighed more (p less than 0.05). Relative testis weights in the short-day group began to decrease (p less than 0.005) within 10 days and continued to decline. Testis homogenate K(+)-pNPPase- (as a measure of Na+,K(+)-ATPase) and Mg(2+)-pNPPase-specific activities closely paralleled testis weight, with the short-day animals (p less than 0.05) differing after 10 days. Plasma testosterone levels remained below adult levels through exposure Day 15, but were relatively lower (p less than 0.05) in the short-day group after 10 days. Spleen weights were similar for the long- and short-day groups. The short-day group had larger thymus weights after 12 days (p less than 0.05), but thymus enzyme activities did not differ between the two groups. We conclude that cation pump activities in the Siberian hamster testis are significantly affected by changes in photoperiod length.     

Differential effects of testosterone and progesterone on the activation and retention of courtship behavior in sexual and parthenogenetic whiptail lizards

Both testosterone (T) and progesterone (P) facilitate the expression of male-typical sexual behavior in a variety of animals, including rodents and lizards. In two species of whiptail lizards, Cnemidophorus inornatus and C. uniparens, both hormones elicit the full repertoire of courtship behavior. However, the relative efficacy of the two hormones is unknown. In Experiments 1 and 2 we assessed differences in capacity of exogenous T and P to induce male-typical courtship behavior in gonadectomized whiptail lizards. In both species, individuals implanted with T showed more frequent courtship behavior relative to those implanted with P or cholesterol. In Experiments 3 and 4 we examined whether T and P differentially affected the retention of courtship behavior following implant removal. In both species, individuals implanted with T showed more courtship behavior following implant removal than those previously given P. In these experiments, implants were removed at a time when individuals in both groups were behaviorally similar; therefore, the differences in behavior following implant removal were not due to differences in the amount of courtship experience. Taken together, the hormone that was more effective at activating courtship behavior was also more effective at maintaining courtship behavior following implant removal. In summary, though both T and P can elicit identical sexual behaviors in both whiptail species, T has a greater and more lasting effect on courtship behavior and possibly on the neural circuits underlying courtship behavior.     

Differential effects of testosterone and dibutyryl cyclic AMP on the meiotic maturation of mouse oocytes in vitro

Fully grown, meiotically immature mouse oocytes were isolated and cultured under varying conditions with the aim of determining a) whether the inhibitory effects of testosterone on oocyte meiotic maturation require the synthesis of new oocyte proteins and b) if the meiosis-inhibiting effects of testosterone and dibutyryl cyclic AMP (dbcAMP) are distinct and can be differentiated. We found that the inclusion of puromycin in culture medium containing testosterone has no effect on the meiosis-inhibiting potency of testosterone or upon the reversibility of testosterone effects. We conclude that testosterone inhibits oocyte meiosis by a mechanism that is independent of protein synthesis. We also found that oocytes exposed to testosterone recover more rapidly, as evidenced by the timing of germinal vesicle breakdown (GVBD) following placement in a control medium, than do oocytes exposed to dbcAMP. Through further investigation of this phenomenon we have determined the sequence of testosterone and dbcAMP effects relative to the time course of GVBD. A testosterone-sensitive event occurs 20 min prior to GVBD, while the dbcAMP-sensitive event precedes GVBD by 41 min. The nature of this difference may involve the differential interaction of testosterone and dbcAMP with a set of puromycin-sensitive proteins that are required for GVBD. When oocytes were initially cultured in medium containing both puromycin and either testosterone or dbcAMP and then moved to medium containing puromycin alone the incidence of GVBD was reduced relative to oocytes never exposed to puromycin. This observation suggests that mouse oocytes contain proteins that are required for GVBD and that experience a high turnover rate. The degree of reduction in GVBD was a function of the length of puromycin exposure and was significantly greater in dbcAMP- than in testosterone-exposed oocytes. If oocytes were initially cultured in medium containing puromycin and dbcAMP, the rate of GVBD upon removal of dbcAMP was initially slow but increased with time. This observation is consistent with the hypothesis that dbcAMP inhibits oocytes at a point prior to the functioning of the puromycin-sensitive proteins. However, if oocytes were cultured in medium containing puromycin and testosterone the rate of GVBD following testosterone removal was not significantly reduced relative to oocytes that were not exposed to puromycin. This observation suggests that testosterone acts to inhibit meiosis at a site beyond the function of the puromycin-sensitive proteins or that testosterone causes a reduction in the turnover rate of these proteins.(ABSTRACT TRUNCATED AT 400 WORDS)     

Stimulatory and inhibitory effects of testosterone on testes in Atlantic salmon male parr

Immature 1-year-old Atlantic salmon Salmo salar parr were implanted with Silastic capsules of different sizes filled with testosterone (T). Testosterone had both positive and negative effects on testicular weights, spermatogenesis and steroidogenesis. The positive effects: higher incidence of males with enlarged gonads, spermiation, and high plasma levels of 11-ketotestosterone (11-KT) and 17,20β-dihydroxy-4-pregnen-3-one (17,20β-P), were most pronounced in males treated with small T capsules. The negative effects: suppression of gonadal development and depressed plasma levels of 11-KT and 17,20β-P compared with mature controls, were most evident in fish treated with large T capsules.     

Ratios of serum concentrations of testosterone and progesterone from yearling bulls with small testes

Thirty crossbred bulls, 12 to 13 mo of age, were used to examine the relationship of testosterone and progesterone concentrations and testosterone: progesterone ratio to measurements of testicular function. Bulls were allotted to 1 of 2 groups based on scrotal circumferences (SC) as follows: the Small SC (n=20) group had scrotal circumference less than 28 cm while the Large SC (n=10) group had scrotal circumference greater than 28 cm. All bulls were administered GnRH (100 mug, im), and blood was obtained immediately prior to injection (t=0), 30 min after injection (t=30) and 2 to 3 h after injection (t=150). Serum was assayed for concentrations of testosterone and progesterone. Semen was evaluated for the percentage of morphologically normal spermatozoa. Testicular parenchyma was sectioned and stained, and 300 cross sections per testis of seminiferous tubules were examined under a light microscope and classified as either active (spermatocytes and spermatids present) or inactive (no spermatocytes or spermatids present). Although progesterone concentrations varied widely (range: 21 pg/ml to 1070 pg/ml), repeated measurements from individual bulls were highly correlated (r(2)=0.74) and did not change significantly (P > 0.1) in response to GnRH treatment. Small SC bulls had a higher percentage of inactive seminiferous tubules (P < 0.001) and a lower percentage morphologically normal spermatozoa (P < 0.001) than Large SC bulls, but no differences in testosterone or progesterone concentrations or in the ratio of testosterone: progesterone were detected. Mean serum testosterone concentration increased (P < 0.0001) by 30 min after GnRH treatment and continued to increase (P < 0.0001) through t=150 but did not differ (p > 0.1) between groups. Normal testosterone secretion in response to GnRH injection suggested that no biochemical lesions in the testosterone production pathway were present in bulls with very small scrotal circumference.     

Influence of melatonin on the testicular regression induced by subcutaneous testosterone pellets in male rats kept in long or short photoperiod

Daily afternoon injections of 25 micrograms melatonin for 12 weeks had no effect on testicular weights of male rats kept in long photoperiod (14L:10D); similarly, exposure of rats to short photoperiod (2L:22D) had no effect on gonadal weight. However, rats maintained in a long or short photoperiod and implanted every 2 weeks with a 15 mm Silastic pellet containing testosterone showed a significant reduction in testicular weight; this effect was more pronounced in rats exposed to a short photoperiod. Melatonin injections in testosterone-treated rats in a long photoperiod exacerbated the inhibitory effects of testosterone alone. Subcutaneous 2-weekly implants of a beeswax pellet containing 1 mg melatonin reversed the effects of the melatonin injections on relative testicular weights but not those due to short photoperiod exposure. Testosterone implants significantly reduced pituitary LH values in long and short photoperiod-exposed animals, more particularly in those exposed to short photoperiod. Melatonin injections alone or in combination with melatonin pellets did not further exaggerate the depression in pituitary LH due to testosterone alone in long photoperiod-exposed animals; similarly melatonin pellets did not reverse the depression in pituitary LH observed. No significant differences in plasma prolactin concentrations or in thyroxine concentrations or free thyroxine index were observed after any combination of treatments. We therefore suggest that the effects observed with short photoperiod may be due to melatonin.     

Pregnenolone and progesterone metabolism by the testes of Bufo arenarum

Sliced testis tissue from Bufo arenarum was incubated in the presence of [³H]pregnenolone. Testis fragments were also used for double isotope experiments using [³H]pregnenolone and [¹⁴C]progesterone. Specific activities were equated with the addition of radioinert pregnenolone. When yields of radiometabolites were analysed, pregnenolone was found to be a good precursor for C₁₉ steroids such as dehydroepiandrosterone, 5-androsten-3β,17β diol, testosterone, 5α-dihydrotestosterone and a C₂₁ steroid, 5α-pregnan-3,20 dione. Progesterone mainly converts to 5α-pregnan-3,20 dione, a steroid with unknown function in amphibians. The 5-ene pathway, including 5-androsten-3β,17β diol as intermediate, could be predominant for androgen biosynthesis. Testes bypass not only progesterone but also androstenedione for testosterone biosynthesis. Accepted: 17 April 1998