Relationship between functional activity of the adrenal glands and gonads in monkeys during puberty]

Testicular and adrenal cortex endocrine functions were simultaneously tested in pubertal baboons (papio hamadryas) during one year. Concentrations of androgens (testosterone, 5 alpha-dihydrotestosterone, dehydroepiandrosterone), corticosteroids (cortisol, progesterone, 17-hydroxyprogesterone, 17-hydroxypregnenolone) and aldosterone were determined in blood of the experimental monkeys by radioimmunoassay. It was shown that with increasing androgen (testosterone and dihydrotestosterone) levels in blood of the pubertal monkeys there was a significant decrease of corticosteroid concentration which was most pronounced in the monkeys with maximum increase of testosterone level. Described hormonal changes did not influence aldosterone concentration.     

Circadian rhythms of androgen levels in the blood of male hamadryas baboons

Radioimmunoassay was used to investigate circadian rhythms of blood plasma testosterone, 5 alpha-dihydrotestosterone, androstendione and dehydroepiandrosterone in intact animals and in those preadapted for experimental conditions. Blood plasma of monkey demonstrated monophasic circadian rhythms in the levels of androstendione, dehydroepiandrosterone and 5 alpha-dihydrotestosterone, with the maximal values seen early in the morning and minimal values in the evening. Circadian rhythms of testosterone were opposite in character. The adaptation of monkeys for housing and experimental conditions was marked by an increase in testosterone and 5 alpha-dihydrotestosterone levels in the blood as well as by an insignificant elevation of the levels of androgens of adrenal origin, leading to the changes in circadian rhythms of steroid hormones.     

Development of the hormonal reaction to stress in the ontogenesis of the baboon Papio hamadryas

The hormonal reaction of adrenal and gonadal glands of baboons at various ages was studied under 2 hr immobilization stress condition. Concentrations of cortisol, 11-deoxycortisol, 17-hydroxypregnenolone, dehydroepiandrosterone and testosterone were determined by radioimmunoassay in the monkey blood plasma at different times during the stress reaction. A more pronounced reaction of adrenal cortex was shown in 1 year old baboons. The peak of cortisol level in immature monkeys under stress conditions was registered much earlier than in adult monkeys.     

5 alpha-dihydrotestosterone administration converts indolamine metabolism and porphyrin content of the female Syrian hamster Harderian gland to the male type

The effects of ovariectomy and exogenous androgen administration on the indole and porphyrin metabolism of Syrian hamster Harderian glands were studied. Ovariectomy alone had no effect on any of the parameters analyzed. The administration of either testosterone or 5 alpha-dihydrotestosterone increased the activity of N-acetyltransferase in the Harderian glands. However, androgen treatment failed to change the activity of hydroxyindole-O-methyltransferase. Melatonin content of the glands dropped 20 days after treatment with testosterone and 10 days after the administration of 5 alpha-dihydrotestosterone. The porphyrin content of the Harderian glands was dramatically depressed after the administration of either androgen. It is concluded that the Harderian glands of Syrian hamsters are under an androgenic control involving 5 alpha-dihydrotestosterone.     

Alteration in the histochemical profile of steroid dehydrogenases in the uropygial gland of male domestic pigeons following androgen deprivation and testosterone supplement

Activities of delta 5-3 beta-hydroxysteroid dehydrogenase (3 beta-HSDH) and 17 beta-hydroxysteroid dehydrogenase (17 beta-HSDH)) were histochemically demonstrated in the alveoli of the uropygial glands of male domestic pigeons (Columba livia Gmelin). The level of 3 beta-HSDH (Substrates: pregnenolone and dehydroepiandrosterone) was very low. On the other hand, an intense activity of 17 beta-HSDH was observed in the glandular alveoli of both sexually immature and adult pigeons when testosterone was used as the substrate. In the adult specimens, bilateral castration resulted in a moderate increase of the level of 17 beta-HSDH indicating a faster rate of conversion of testosterone into the weaker androgen, namely, delta 4-androstene-3,17-dione. On the other hand, administration of testosterone propionate (TP), 500 micrograms/bird/day for 15 days in the intact prepubertal and castrated adult pigeons caused a perceptible depletion of 17 beta-HSDH, which may be correlated with the increase in the local concentration of testosterone. Low dose of TP (100 micrograms/day) had no effect. When estradiol-17 beta was used as the substrate for 17 beta-HSDH, there was only a feeble response within the alveoli.     

Response of the epididymis, ductus deferens & accessory glands of the castrated prepubertal rhesus monkey to exogenous administration of testosterone or 5alpha-dihydrotestosterone.

The response of the epididymis, ductus deferens, and accessory glands of the castrated prepubertal rhesus monkey to exogenous administration of testosterone or 5alpha-dihydrotestosterone (DHT) was investigated. 200 or 800 mcg of either steroid/day were administered for 60 days beginning on the day after castration. Castration caused a marked regression of the weight of and secretory function of the reproductive organs; testosterone/DHT stimulated their growth and secretory activity which were maintained at the level of the controls. The weight of the caput epididymides however, was unaffected by testosterone but was stimulated by DHT. DHT caused a greater stimulation of the growth and secretory activity of the reproductive organs than testosterone and also caused a hyperstimulation of secretion by the seminal vesicles. The data, analyzed statistically, show that the accessory organs of the prepubertal rhesus monkey are affected by castration and vary in their response to stimulation by exogenous androgens.     

The effect of ACTH therapy on serum dehydroepiandrosterone, androstenedione, testosterone and 5 alpha-dihydrotestosterone in infants

Serum concentrations of dehydroepiandrosterone (DHEA), androstenedione, testosterone, 5 alpha-dihydrotestosterone and cortisol were measured in 10 infants (age 5-22 months) before, during and after 6-weeks of ACTH therapy for infantile spasms. During therapy, their mean DHEA concentrations increased 2.3-fold, androstenedione 12.3-fold, testosterone 2.7-fold, 5 alpha-dihydrotesterone 2.5-fold and cortisol 2.9-fold compared to pre-therapy values. Serum dehydroepiandrosterone sulphate (DHEA-S) concentrations were also increased during ACTH therapy above the normal prepubertal range. Three days after the cessation of ACTH treatment, all androgens had returned to the pre-therapy level. We conclude: At least in pharmacologic doses ACTH alone stimulates adrenal androgen secretion in infants, excluding the necessity of a separate adrenal androgen stimulating hormone.     

Androgenic control of N-acetyltransferase activity in the harderian glands of the Syrian hamster is mediated by 5 alpha-dihydrotestosterone

N-acetyltransferase (NAT) activity in the Harderian glands of intact and gonadectomized male and female Syrian hamsters was evaluated. The exogenous administration of 5 alpha-dihydrotestosterone (DHT) to castrated males and intact females produced an increase in NAT values, which reached the values present in the glands of intact males. The administration of a 5 alpha-reductase inhibitor to intact males led to a decrease in NAT activity, suggesting that testosterone is converted in DHT within the glands. It is concluded that NAT activity in the Syrian hamster Harderian glands is under androgenic control, the active steroid being DHT.     

Plasma concentrations of testosterone, dihydrotestosterone, delta 4-androstenedione, dehydroepiandrosterone and oestradiol-17 beta in the crab-eating monkey (Macaca fascicularis) from birth to adulthood

Plasma testosterone, 5 alpha-dihydrotestosterone (DHT), delta 4-androstenedione, dehydroepiandrosterone (DHA) and oestradiol-17 beta concentrations of crab-eating macaques after birth were analysed by RIA. The profiles of plasma testosterone and DHT exhibited four phases: (1) a neonatal phase (0 to 3-4 months of age) with considerable synthetic testicular activity; (2) a phase of 'infancy' (generally up to 29 months of age) during which the values of both androgens were low; (3) a prepubertal phase (generally up to 43 months of age) when circulating values oscillated with wider individual variations, and (4) a pubertal phase when the concentrations increased in parallel and concomittantly with the onset of meiosis and the establishment of spermatogenesis. The testosterone values continued to increase, reaching adult values at about 5-6 years of age, whereas DHT levels tended to stabilize from 4-5 years. Relatively high androstenedione values during the neonatal phase decreased progressively until puberty, then increased again slowly up to the adult stage when they plateaued at about neonatal levels. The DHA levels were high during the first months, decreased at about 1 year, remained stable during infancy and prepuberty and then declined again during puberty. At about 5 years, the values were 28% of those in neonates. There was no evidence of an adrenarche before the first signs of sexual maturity were observed. Oestradiol-17 beta concentrations were high at birth and until 3 months, then decreased and remained steady from 1 year of age until adulthood, except at the onset of puberty (27-30 months of age) when high values were again noted. Our results show that, during the neonatal period, the testis exhibited considerable secretory activity.     

Intratesticular steroid levels and their hormonal control

Litter-mate adult male rats were treated with daily intramuscular injections of ACTH (10.5 micrograms), dexamethasone (2.0 mg), ethynyl estradiol (1.7 micrograms) and hCG (5 IU) for three consecutive days. The animals were sacrificed on the fourth day and the intratesticular and peripheral plasma steroid levels were analyzed. The steroids measured by radioimmunoassay included pregnenolone, 17-hydroxypregnenolone, dehydroepiandrosterone, progesterone, 17-hydroxyprogesterone, androstenedione, testosterone and dihydrotestosterone. In addition, the sulphoconjugated forms of pregnenolone, dehydroepiandrosterone, testosterone and dihydrotestosterone were estimated in the peripheral blood. The administration of ACTH diminished the intratesticular levels of all steroids studied. Also dexamethasone and ethynyl estradiol treatment suppressed all intratesticular steroid levels, except that of pregnenolone (the former) and of 17-hydroxyprogesterone (the latter). The suppressive effect of ethynyl estradiol was strongest on the levels of the delta 5-steroids and that of dexamethasone on the delta 4-steroids; the latter was significantly stronger than the effect of ACTH. The stimulatory effect of hCG was limited to the metabolism of progesterone and was restricted to the sequence: 17-hydroxyprogesterone----androstenedione----testosterone---- dihydrotestosterone. Dexamethasone-suppression, and hCG-stimulation of the intratesticular levels of delta 4-steroids, was mirrored by corresponding changes in the peripheral plasma levels, with the exception of the plasma levels of androstenedione which were not influenced by any of the treatments studied. Also the suppression of intratesticular testosterone and dihydrotestosterone levels by ACTH, dexamethasone, or ethynyl estradiol was closely reflected by their plasma levels both in the unconjugated and sulphoconjugated forms. On the hand, the administration of ACTH diminished the intratesticular levels of pregnenolone and progesterone but significantly increased those in the plasma. Moreover, both ACTH and ethynyl estradiol reduced the levels of all delta 5-steroids in testicular tissue, but not in the peripheral plasma, although they decreased the circulating levels of pregnenolone sulphate and dehydroepiandrosterone sulphate. The data are interpreted as suggesting that the hormonal agents studied interfere with testicular steroidogenesis through different mechanisms.     

Effects of hCG on serum levels of testosterone, dihydrotestosterone and androstenedione in male mice

The concentrations of testosterone (T), 5alpha-dihydrotestosterone (5alpha-DHT) and delta4-androstenedione (delta4 A) in the circulating blood were increased by a single subcutaneous injection of 10 IU of hCG in the mouse. The response of androgen synthesis to hCG stimulation was rapid and persisted over a period of 96 h. The increased supply of T to the peripheral organ(s) under the stimulation of hCG might contribute to an increase in circulating 5alpha-DHT. However, the changes in T/5alpha-DHT ratio implies that the stimulation of T and 5alpha-DHT production by hCG were not of the same degree.     

Identification of receptors in a system of functionally heterogeneous proteins specifically bound to androgens in rat liver cytosol

The methods of androgen receptor (RA) isolation and identification in rat liver cytosol were studied. It was shown that male rat liver contains a system of specific androgen (A)-binding proteins consisting of at least three main components: RA, delta 4-androstendione (delta 4-A)-binding component and an unusual estrogen-binding protein interacting also with A and the first two components in females. The identity of one of A-binding components to RA was proved by cumulative properties of this component which are similar to those of RA from other tissues. These properties are as follows: 1) high values of apparent association constant, Ka, for 3H-R1881 (2.8 +/- 0.3 X 10(8) M-1) and 3H-5 alpha-dihydrotestosterone (3H-DHT) (5.0 +/- 0.4 X 10(8) M-1); 2) low binding capacity--approximately 10 fmol/mg of protein of nonfractionated cytosol; 3) pronounced specificity of affinity for active A (DHT, R1881, testosterone); 4) large size of the protein molecule (6.5 +/- 0.25 nm); 5) ability to decrease this size to 3.2 +/- 0.08 nm in a high ionic strength buffer; 6) precipitation at low concentrations of ammonium sulfate: 7) strong interaction with heparin-Sepharose. The properties of the delta 4-A-binding component do not coincide with those of RA: it has a low Ka for 3H-delta 4-A (1.15 +/- 0.5 X 10(6) M-1), a high binding capacity (1.22 +/- 0,12 pmol/mg of protein of nonfractionated cytosol) and can bind various delta 4-3-ketosteroids irrespective of the degree and nature of their biological activity. It was concluded that preliminary isolation of rat liver RA on heparin-Sepharose can be used for differential identification and characterization of this protein.     

In vitro metabolism of C19 steroids in human endometrium

In order to quantitate the extent of intracellular metabolic conversions of C19 steroids in human endometrium, specimens of proliferative and secretory tissue were superfused at a constant rate with several pairs of labeled compounds at low concentrations. About 16% of dehydroepiandrosterone sulfate interacting with endometrial cells was converted to dehydroepiandrosterone and about 3% of this compound was converted to androstenedione. Androstenedione was reversibly reduced to testosterone and the extent of this conversion was shown to be several-fold higher in secretory than in proliferative tissue. About 1% of testosterone entering the cells was reduced to 5 alpha-dihydrotestosterone. These results demonstrate that the conversion of the main circulating C19 steroids in women, i.e. dehydroepiandrosterone sulfate and androstenedione, to 5 alpha-dihydrotestosterone, the compound considered to be the true intracellular androgen, is very small. In contrast, formation of testosterone from androstenedione is extensive and increases during the luteal phase under the influence of progesterone, a hormone known to stimulate the activity of 17 beta-hydroxysteroid dehydrogenase in human endometrium.     

Possibility of adrenal-testicular interaction as indicated by plasma androgens in response to HCG in men with normal, suppressed and impaired adrenal function.

In order to assess possible adrenal-testicular interactions in vivo, adrenal and testicular plasma androgens (testosterone, delta4-androstenedione, dehydroepiandrosterone and dehydroepiandrosterone sulfate) were measured by specific radioimmunoassays before and after stimulation with HCG in men with normal, dexamethasone suppressed and impaired adrenal function. It was found that men with Addison's disease, in whom circulating dehydroepiandrosterone sulfate levels amounted to 1/10 of normal values, had a decreased response of testosterone to HCG. Simultaneously, the Addison patients and the men under dexamethasone had only an increase of delta4-androstenedione but not of dehydroepiandrosterone, while normal men showed an almost equal increase in both precursors under HCG. The results are interpreted as demonstrating that the delta5-pathway in the testis becomes less important under adrenal suppression and in Addison's disease due to a lack of substrate (possibly dehydroepiandrosterone sulfate) from the adrenals.     

Testicular endocrine function in hamadryas baboons (Papio hamadryas) during chronic emotional stress

Radioimmunoassay was applied to examine the time course of blood plasma testosterone content in 10 adult male hamadryas baboons under repeated immobilization stress (IS). In the first series of experiments, the monkeys (5) were subjected to daily IS for 2 h during 6 days. In the second series, 5 animals were exposed to 2-hour immobilization 3 days prior to the analogous stressful cycle. In response to intermittent IS the first series males demonstrated profound and long-term suppression of blood testosterone that persisted for 2 days after IS cessation. Preliminary application of a stressful stimulus substantially reduced the degree of blood testosterone suppression. Transitory decrease in blood testosterone was observed 2 hours after the outset of each stressful exposure, followed by complete recovery of testosterone blood level after 6 hours. The differences in the time course of testosterone indicate that the response of the pituitary-gonadal system is dependent on the regimen of stressful stimulus application.     

Peripheral aromatization: studies on controlling factors

Using constant infusions of 3H-labeled androgens and 14C-labeled estrogens with measurements of radiolabeled estrogens in blood and/or urine we have carried out studies on the peripheral aromatization of androgens in humans, nonhuman primates, sheep, and rabbits. In the human, aromatization is increased in women as they become postmenopausal, although the mechanism remains uncertain. In humans and cynomolgus monkeys the administration of ACTH and/or glucocorticoids does not increase peripheral aromatization, but results in a slight decrease in the aromatization of androstenedione. The administration of l-thyroxine to cynomolgus monkeys increases peripheral aromatization of androstenedione from basal, 1.16 +/- 0.15%, to 1.71 +/- 0.14% probably due to increased tissue blood flow. The aromatization of testosterone is not affected, probably due to an increase in sex hormone-binding globulin. Peripheral aromatization occurs to a similar degree in humans, rhesus and cynomolgus monkeys, and baboons, but is much lower in sheep and rabbits. The compound 10-(2-propynyl)-estr-4-ene-3,17-dione is an effective inhibitor of the peripheral aromatization of both androstenedione and testosterone.     

Involvement of dietary glucose in adrenal steroidal regulation of spermatogenic and steroidogenic activities in prepubertal rat testis.

Effects of dietary and supplemented dextrose energy on the role of corticosterone (Comp. B) or dehydroepiandrosterone (DHA) in spermatogenic and steroidogenic activity in the bilaterally adrenalectomised prepubertal rat testis were studied. Adrenalectomy reduced the body and testis weight, numbers of the stage VII cell types [spermatogonia A (A), preleptotene (PL) and pachytene (P) spermatocytes and step 7 spermatid (7)], testicular delta 5-3 beta-hydroxysteroid dehydrogenase (delta 5-3 beta-OH-SDH) activity and serum testosterone. Adrenalectomy also caused reduction of energy intake due to loss of appetite which was stimulated by hormone replacement therapy. Treatment of adrenalectomised rats with DNA or corticosterone enhanced the spermatocyte population and the enzyme activity, especially after 30 days of age. Dextrose supplementation with hormone treatment however, did not produce significant additive effect on stage VII cell counts, but delta 5-3 beta-OH-SDH activity showed additive effect in this age group. Results suggest that adrenal steroids regulate testicular steroidogenesis and spermatogenesis during the prepubertal ages by modifying the supply of dietary glucose.     

Changes in plasma steroid levels after single administration of hCG or LHRH agonist analogue in dog and rat

The present study was designed to investigate the effect of acute administration of gonadotropin on testicular steroid secretion in dog and rat. Animals received a subcutaneous injection of 25 IU/kg of hCG or 1.5 microgram/kg of [D-Trp6, des-Gly-NH2(10)]LHRH ethylamide (LHRH-A). Testosterone is the predominant steroid measured, in dog plasma, under basal conditions. After LHRH-A injection, testosterone levels are not significantly changed while dehydroepiandrosterone and androst-5-ene-3 beta,17 beta-diol (delta 5-steroids) levels are stimulated by almost 20-fold (P less than 0.01). When dogs were injected with hCG, we also observed a marked stimulation of dehydroepiandrosterone levels (20-fold; P less than 0.01) accompanied by a small increase of plasma testosterone concentration (2-fold, P less than 0.01). In rats injected with either hCG or the LHRH analogue, an increment of plasma testosterone (7-fold, P less than 0.01) is detected in the first hour while plasma dehydroepiandrosterone levels are slightly stimulated. Moreover, in rats injected with hCG, low plasma steroid levels are present between 4-12 h after injection due to testicular desensitization. This marked decrease is then followed by a second peak of steroid secretion 24 h later. Acute testicular steroidogenic responsiveness to hCG on the dog is, however, different: after stimulation, the levels of plasma dehydroepiandrosterone are maintained at a plateau and slowly decline after 24-48 h. Our data indicate that in dogs, stimulation of testicular steroidogenesis leads to an increase of plasma delta 5-steroid levels while the same stimuli cause, in the rat, a stimulation of delta 4-androgen, particularly testosterone.     

Subnormal pubertal increases of serum androgens in Turner's syndrome

60 patients (139 blood specimens) with Turner's syndrome were investigated in order to obtain information concerning the origin of the increments of androgens during puberty. The concentrations of serum FSH, LH, estradiol, testosterone, 5 alpha-dihydrotestosterone, dehydroepiandrosterone, progesterone, 17-hydroxyprogesterone and pregnenolone in patients less than 10 years old were identical to those previously found in normal healthy girls of the same age. Hence, in adrenarche the early increase of androgen secretion is independent of gonadal hormone secretion. The later increases in serum testosterone and androstenedione in our patients were very small, and the age of 15 years, their concentrations were 50 and 60%, respectively, of the corresponding levels in normal girls of the same age. After 13 years of age, the mean serum dehydroepiandrosterone concentration was also slightly, but significantly (20-30%), lower than in normal girls of the same age. It is concluded that the ovaries are responsible for most of the pubertal rises in circulating testosterone and androstenedione, and possibly for a small part of the late pubertal rise in dehydroepiandrosterone.     

Testicular responsiveness to human chorionic gonadotrophin during transient hypogonadotrophic hypogonadism induced by androgenic/anabolic steroids in power athletes

Serum concentrations of testosterone, 17-hydroxyprogesterone, estradiol and several other unconjugated and sulphated steroids were analyzed before and after a single dose of hCG in 6 power athletes, who had used high doses of testosterone and anabolic steroids for 3 months. The study was carried out 3 weeks after cessation of drug use, but the study subjects were still characterized by hypogonadotrophic hypogonadism. The mean concentrations of serum LH and FSH were 2.6 +/- 0.3 and 1.1 +/- 0.03 mIU/ml (mean +/- SEM), respectively, and the concentrations of several precursors and metabolites of testosterone were lower than those before drug use. In contrast, circulating concentrations of steroid sulphates were not decreased, with the exception of dehydroepiandrosterone sulphate. After hCG injection serum testosterone and 5 alpha-dihydrotestosterone concentrations increased significantly, whereas no increases in estradiol and 17-hydroxyprogesterone concentrations were observed. These results demonstrate that during transient hypogonadotrophism in adult men, the testicular responsiveness to a single injection of hCG is similar to that in prepubertal boys without any sign of steroidogenic lesion at the 17,20-desmolase step. Therefore, the appearance of the possibly estradiol-mediated inhibition at the level of C21-steroid side-chain splitting in testosterone biosynthesis seems to be dependent on priming by gonadotrophins.