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Leuprolide (Lupron, TAP Pharmaceuticals), a potent agonist analogue of GnRH, has been shown to suppress testicular androgen production in animals. In order to determine the potential of leuprolide as an alternative to surgical castration in human males with prostatic cancer, this agent was administered to castrate and noncastrate males with carcinoma of the prostate. Baseline and treatment levels of LH, FSH, testosterone and dihydrotestosterone were determined serially. Leuprolide suppressed pituitary production of LH and FSH and, consequently, testicular production of testosterone and dihydrotestosterone. This agent simulates the results achieved with surgical castration.

Alterations in the metabolism of testosterone (T) and dihydrotestosterone (DHT) induced by diethylstilbestrol (DES) or medroxprogesterone acetate (MPA) could account for the beneficial therapeutic effect of these agents in prostatic carcinoma. To investigate this possibility we sutdied plasma kinetics of T and DHT in 17 elderly patients with prostatic carcinoma, before and after treatment with DES (1 or 5 mg/d) or MPA (10 or 30 mg/d) for 30 days. Metabolic clearance rates (MCR) were determined with the single injection technique and by use of two compartment model, plasma concentrations (PC) of T and DHT by radioimmunoassay, the per cent of T bound to plasma protein (T-binding) by charcoal adsorption of the unbound steroid. Production rate (PR) and PC of T were lower, PR and PC of DHT were higher in our patients than in normal men. With both DES regimens, PR, PC and MCR of either androgen declined; however, T was suppressed to a much greater extent than DHT. In either instance, the decrease may have been caused by direct suppression of testicular androgen synthesis and/or by decreased gonadotropin stimulation. Enhanced T-binding played an additional role in reducing the free testosterone index. High and low dose of DES were equally effective. The low dose regimen of MPA did not influence androgen metabolism. MPA in the higher dose suppressed PR and PC of T and DHT, possibly due to effects on testicular synthesis or by gonadotropin suppression as suggested for DES. In contrast to DES, MPA failed to cause profound changes in MCR of either androgen or in T-binding. When judged by its influence on the metabolism of T and DHT in prostatic carcinoma, MPA in higher doses is much less effective than either dose regimen of DES.

In quail, the hypothalamus enzymatically transforms testosterone (T) into estradiol (E2), 5 alpha-dihydrotestosterone (5 alpha-DHT), and 5 beta-dihydrotestosterone (5 beta-DHT). During the embryonic life, the 5 beta-reductase activity is very high, which probably protects the brain of males from being behaviorally demasculinized by their endogenous T. 5 beta androstanes are inactive androgens. The decrease of 5 beta reductase with age during sexual maturation corresponds to a potentiation of the effects of T as shown by experiments that compared the effects of T and 5 alpha-DHT in adult and young quail. T metabolism is also involved in the activation of male behavior in the adult. T aromatization is probably essential for behavioral activation, but nonaromatizable androgens such as methyltrienolone, and to some extent 5 alpha-DHT, can also stimulate sexual behavior in castrates. These enzymatic activities show a clear neuroanatomical localization and are sexually dimorphic. Males produce more active metabolites (E2, 5 alpha-DHT) than females, which could explain the male's greater sensitivity to T treatments. It thus appears that T metabolism is involved in the differentiation and activation of behavior in quail.

Numerous investigators demonstrated increased 5 alpha-reductase activity in testes of developing rats. The rapid in vitro metabolism of progesterone to 5 alpha-reduced androgens occurs at certain stages of testicular development. This was considered evidence for the conclusion that testosterone is primarily an intermediate rather than the final product in testes of immature rats. However, a discrepancy is noted when developmental patterns of circulating or intratesticular levels of androgens are compared with the patterns of accumulation of metabolites of progesterone in vitro. In blood and testicular tissue of 17-20 day old rats a testosterone peak has been reported, while in the in vitro studies such peak was not observed. In this study radiolabelled pregnenolone was utilized in vitro as a substrate, and a pattern of androgen formation similar to that observed in the in vivo studies was noted. A peak of androgen formation (testosterone and 5 alpha-androstanediol) was observed used prior to completion of the meiotic prophase. However, when testosterone was utilized as the substrate, no correlation between 5 alpha-reduction and completion of the meiotic prophase was detected. This suggests that the rise in testosterone may be associated with completion of the meiotic prophase. Investigation of the androgen metabolic pathways revealed the following age-related patterns: no change in 17 beta-hydroxysteroid dehydrogenase, activation of 5 alpha-reductase at 12 days of age, and activation of 3 alpha-hydroxysteroid dehydrogenase between 14 and 18 days of age.(ABSTRACT TRUNCATED AT 250 WORDS)

In an infant with gonadal dysgenesis and somatic anomalies, the internal and external genitalia were female but the gonads contained tubular structures suggesting male differentiation. The karyotype was 46,XY with no evidence of structural aberration or mosaicism. Hormonal metabolism and H-Y antigen expression were assayed in cultured gonadal cells. Although unable to synthesize testosterone, the cultured cells were able to convert it to dihydrotestosterone. H-Y antigen was present, perhaps at a level lower than that in cells from normal XY males. Our observations indicate that a modicum of testicular organogenesis may precede the involution that results in a streak gonad in some cases of gonadal dysgenesis.

Although androgens have been implicated in follicular atresia, ovarian follicular androgen synthesis is required for preovulatory follicular growth. To localize the site(s) of androgen biosynthesis and to obtain a better understanding of the regulation of the androgenic pathway(s) in rat ovarian follicles we examined the relative abilities of developing follicles to accumulate specific androgens [testosterone (T) and dihydrotestosterone (DHT)] using both radioimmunoassay (RIA) and 3H-substrate metabolism techniques. Small antral and preovulatory follicles were obtained from control or human chorionic gonadotropin (hCG)-primed immature rats, respectively (Richards and Bogovich, 1982). Small antral follicles, theca and granulosa cells produced little immunoassayable androgen (T + DHT) when incubated with or without 8-bromo-cAMP. In contrast, preovulatory follicles and theca produced more androgen than small antral tissues and in a manner acutely stimulable by cAMP. Granulosa cells produced little androgen under these conditions. Inclusion of [3H] androstenedione in the incubates yielded increased accumulation of [3H] T and [3H] DHT for all small antral and preovulatory tissues. Indeed, granulosa cells from both small antral and preovulatory follicles possessed a remarkable ability to accumulate [3H] T. This ability was not altered by hypophysectomy or subsequent treatment with estradiol and/or follicle-stimulating hormone (FSH). These results suggest that 17-ketosteroid reductase may be a constitutive enzyme in granulosa cells.

The comparative ability of granulosa cells and theca of the hamster preovulatory follicle to produce androgens in vitro from endogenous and exogenous substrates was assessed. The results indicate that theca are the major source of follicular androstenedione, but that the granulosa cells may be the major source of follicular testosterone. Theca and granulosa cells accumulate comparable amounts of dihydrotestosterone from exogenous androstenedione and testosterone and both may be a significant source of follicular DHT. LH stimulates the conversion of progesterone and 17 alpha-OH progesterone to androstenedione, testosterone and DHT in theca. LH does not stimulate the conversion of androstenedione to testosterone or DHT, and that of testosterone to DHT in either granulosa cells or theca. FSH, in granulosa cells but not in theca, stimulates the conversion of adrostenedione to testosterone but it has no effect in DHT accumulation from exogenous testosterone.

Tritiated 5alpha-androstane-3alpha,17beta-diol (3alpha-diol) and 5alpha-androstane-3beta,17beta-diol (3beta-diol) respectively were administered to patients with benign prostatic hypertrophy (bph) undergoing prostatectomy. In prostate and skeletal muscle homogenates and in plasma the total radioactivity content as well as the formation of metabolites were measured. Histological examination of each ectomized prostate was performed to evaluate the cellular composition of the tissue. After 3alpha-diol injection, a higher uptake of radioactivity in the prostate was obtained than after 3beta-diol. Within 30 min the 3alpha-isomer was very efficiently converted to 5alpha-DHT, while most of the 3beta-isomer remained unchanged. There was, however, also after administration of the 3beta-diol a substantial biconversion to 5alpha-DHT as has been confirmed by recrystallization to constant specific radioactivity. Only after 3beta-diol epiandrosterone was detected in small but significant amounts. 3alpha-diol administration resulted in distinct concentrations of 3beta-diol, whereas the conversion of 3beta-diol to the 3alpha-isomer was insignificant. When comparing the histological composition of the prostatic tissue with the accumulation of radioactivity and the formation of metabolites only a weak correlation between glandular structure and radioactivity uptake after 3alpha-diol administration could be revealed.

After the rat preputial gland was treated with collagenase and trypsin, five bands of cells were isolated by centrifugation in Ficoll gradients. Homogenates of the heavier cells (Bands IV and V) which contained less lipids, were more active than the homogenates of the lighter cells (Bands I, II and III) in transforming [1,2-3H]-dehydroepiandrosterone ([1,2-3H]-DHA) into [3H]-androstenedione and [3H]-testosterone and the latter into [3H]-dihydrotestosterone (DHT). In the presence of NAD, NADH and NADPH-generating system, [1,2-3H]-DHA was transformed into [3H]-DHT in 50-60% yield by homogenates of cells in Bands IV and V. DHT levels in the preputial gland were measured by radioimmunoassay. The levels in female rats reduced by 77% from 3.14 +/- 0.27 to 0.72 +/- 0.10 pg/mg tissue after adrenalectomy, and by 45% to 1.71 +/- 0.10 pg/mg tissue after ovariectomy. In male rats, the level reduced by 15% from 4.58 +/- 0.55 to 3.88 +/- 0.62 pg/mg tissue after adrenalectomy and by 40% to 2.74 +/- 0.21 pg/mg tissue after orchidectomy. These results demonstrated the transformation of DHA into DHT in the preputial gland of the rat, and that the adrenal is an important source of precursor steroid (DHA) for DHT formation in the preputial gland.

Reproductive capability requires synchronization of both endocrine and behavioral components of reproduction. The classic view of reproductive behavior was that there was simply direct stimulation of a behavioral response by the appropriate gonadal steroid. However, it has become clear with recent developments in the field of behavioral endocrinology that the relationship of hormonal and behavioral processes is complex. In addition to the complexity of the mechanisms involved, other factors, both social and environmental, influence both the endocrine and behavioral responses. This paper provides an overview of selected issues within the field of behavioral endocrinology which deal with mechanisms of hormonal induction of behavior at various stages of the life cycle and with factors that interact with these processes.

The properties of 5 beta-reductase, 5 alpha-reductase and aromatase, 3 testosterone metabolizing enzymes, were studied in the quail brain by an in vitro incubation technique. The results describe the changes in time of metabolite production and the effects of temperature, enzyme and cofactor concentrations. The apparent Km and Vmax were evaluated for the 3 enzymes. Aromatase and 5 alpha-reductase have a higher affinity but a lower capacity than 5 beta-reductase. The kinetics of the latter enzyme are complex and suggest the presence of two types of enzymes. These characteristics fit in well with the role probably played by the enzymes in vivo.

Male pseudohermaphroditism can be caused by absent müllerian regression, inadequate synthesis of testosterone, inadequate synthesis of dihydrotestosterone, or androgen receptor deficiency. Indications for gender assignment according to the differential diagnosis of male pseudohermaphroditism are suggested.

The effectiveness of aminoglutethimide as an adrenal inhibitor has been well-documented by decreases in plasma testosterone and delta 4 levels, which fall significantly following the drug in previously orchiectomized patients. The use of cortisone or cortisol along with aminoglutethimide complicates the interpretation of the role of aminoglutethimide in effecting clinical responses. However, since physiologic replacement doses were used in most cases, a significant role for cortisone in effecting a clinical response is unlikely. Aminoglutethimide does have side-effects including rash and lethargy. It requires administration of replacement doses of cortisone and sometimes mineralocorticoid as well since it inhibits adrenal steroid synthesis in all pathways. Peripheral adrenal androgen inhibitors, such as flutamide, Megace, cyproterone acetate or 5 alpha-reductase inhibitors, in the future may be equally effective and simpler to administer than aminoglutethimide but objective and adequate numbers of studies using acceptable objective criteria must be done in order to adequately compare these drugs to aminoglutethimide. There appears to be approximately a 33% response rate (partial objective regression and objectively stable) following blockade of adrenal androgens in patients in relapse after castration. Blockade of adrenal androgen is certainly more tolerable and has many fewer side-effects than the alternative of chemotherapy which does not give response rates in most cases that are significantly different from those noted with aminoglutethimide. Murray's paper, combined with prior studies by Drago et al., goes a long way in establishing adrenal androgen blockade with that drug as the next step to be taken in patients following relapse from prior castration (medical or surgical). The most important question revolves around the timing of adrenal androgen blockade. As stated by Murray, will adrenal androgen blockade provide better survival if given earlier following relapse? The answer is not known yet. The answer may come from the work of Labrie [1], Geller and Albert [2] and others, who suggest that total survival in prostate cancer may be improved with blockade of adrenal androgens not after relapse following castration, but with panandrogen blockade at the time of initial therapy for prostate cancer.

Thecal preparations from medium-sized procine ovarian follicles (3.5-5 mm diameter) were incubated for 4 h in a chemically defined medium in the presence or absence of highly purified luteinizing hormone (LH) and/or estradiol-17 beta (estradiol). LH (1 microgram/ml) stimulated the thecal production of testosterone (T) and dihydrotestosterone (DHT) by 2- to 3-fold. Although estradiol (10 microgram/ml) alone had only a slight but non-significant inhibitory effect on basal testosterone production, it significantly inhibited the production of both T and DHT as well as decreasing the DHT/T ratio in a dose-related manner in the presence of LH. Production of cyclic adenosine 3',5'-monophosphate (cAMP) and progesterone by the thecal cells was stimulated 50-to 200-fold and 2.5-fold, respectively, by LH. Estradiol had no significant effect on thecal cAMP and progesterone production in the presence or absence of the gonadotropin. These findings are consistent with the concept that estradiol produced by granulosa cells following hormonal stimulation may serve as a local negative feedback mechanism to control thecal androgen production.

Hormonal regulation of testicular descent has been investigated. Based on experimental studies using the rat and on a review of clinical material it has been determined that testicular descent is an androgen-mediated event directly under the regulation of pituitary gonadotropin. Furthermore, the active androgenic metabolite involved in this process appears to be dihydrotestosterone, which is synthesized by the testis and must be present in high local concentrations to be effective. In addition to these hormonal requirements the anatomy of the inguinal canal must provide unimpeded migration of the testis into the scrotum. Any imbalance or anomaly of these prerequisites may lead to cryptorchidism.

Activities of 5 alpha, 3 alpha, and 3 beta reductases were studied with the use of microsomal fractions isolated from the ovaries of immature female rats after treatment with pregnant mares' serum (PMS) followed by human chorionic gonadotropin (hCG). The effect of exogenous prolactin treatment was similarly assessed by comparing enzymic activity in prolactin-treated rat ovaries to that in control rat ovaries, after pretreatment with PMS. With the use of 3H-testosterone as substrate, reductase activities were determined by measuring the production of dihydrotestosterone and the 3 alpha- and 3 beta-epimers of androstanediol. The rate of formation of tritiated androstanediol from radiolabeled testosterone was ten times higher than that of tritiated dihydrotestosterone during the follicular phase. After ovulation, the metabolism of testosterone to both androstanediol and dihydrotestosterone decreased markedly. Negligible amounts of estrogen and androsterone were formed, and most of the androstanediol was found to be 5 alpha-androstane-3 beta, 17 beta-diol. The Michaelis-Menten constant (Km) for 5 alpha-reductase was 1.3 x 10(-5)M. During the follicular phase, the sum of 3H-dihydrotestosterone and 3H-androstanediol produced from radiolabeled testosterone was found to be 30% greater in prolactin-treated animals than in control animals. Implication of increased ovarian androgen metabolism as a consequence of hyperprolactinemia are discussed.

The ongoing development and gradual availability of the new anti-androgens hold exciting clinical implications for the future. The biosynthesis and interchangeability of the sex steroids, the roles of the ovaries and adrenals and the value and interpretation of biochemical assays in clinical practice are briefly discussed. Because the anti-androgens are used primarily for seborrhoea/acne/hirsutism/oligomenorrhoea, the pathophysiological basis of this socially debilitating syndrome is discussed. The classification of the anti-androgens, their indications, side-effects, dosage schemes and results of treatment are reviewed. Finally, a summary of a possible clinical management regimen is presented.

Testosterone secreted from the testis is converted to dihydrotestosterone (DHT) in target tissues by 5 alpha-reductase and DHT exerts its action on the nucleus. Since skin is one of the target tissues of androgen we established a system for culturing human skin fibroblasts, and we investigated the conversion of testosterone to DHT in genital and nongenital skin fibroblasts obtained from males as well as from females. In our system, fibroblasts were incubated with 0.1 microM 1, 2-3H testosterone, and labeled metabolites were separated by thin layer chromatography. DHT formation was linear for 6 hours and the rate of DHT formation correlated with the growth phase of the fibroblasts. In the course of serial subculture, DHT formation did not change between the 3rd and 10th subculture in four cell strains. DHT formation in nongenital skin fibroblasts did not differ significantly between males and females. Fibroblasts obtained from male genital skin produced significantly greater amounts of DHT than those from male or female nongenital skin. Furthermore, DHT formation in genital skin fibroblasts did not differ between normal males and hypogonadal males. These data suggest that genital skin is the target tissue where testosterone strongly converts to DHT and the difference of male secondary sexual characteristic appearance between males and females may not depend on the sensitivity of the target tissues to androgen but on the secretion rate of androgen.

Human pubic skin was obtained from normal subjects and patients with abnormal sex differentiation. Skin samples (200 mg) supplemented with NADPH, were incubated for 1 h with labelled testosterone. The conversion of testosterone to dihydrotestosterone, 3alpha- and 3beta-androstanediol was calculated. This conversion averaged 14.9 plus or minus 3.4% (SE) in 11 normal men and 3.6 plus or minus 1.4% (SE) in 8 normal women. In 4 children as in 4 young hypogonadotrophic hypogonadal men, the conversion rate of testosterone to 5alpha-reduced metabolites was low (0.8 to 3.5%) and increased at puberty (13.5 to 19.2%). After administration of HCG for 3 months to 1 of the hypogonadal men, it reached 30.2%. Inversely, the formation of dihydrotestosterone and androstanediols from testosterone was suppressed in 2 men treated with large doses of oestrogen. In 3 subjects with an incomplete form of testicular feminization syndrome, the conversion rate of testosterone to 5alpha-reduced metabolites was in the normal male range (6.4 to 18.3%), whereas it was low in one case of the complete form of the syndrome (1.5%). In 9 women with idiopathic hirsutism the rate of 5alpha-reduced metabolites recovered from testosterone was close to that of normal men (13.5 plus or minus 5.5% (SE). From these results, it is postulated that in human subjects, there is a good correlation between hair growth in skin from a sexual area and the extent of testosterone 5alpha-reduction in this tissue. Such an enzymatic activity might be induced by active androgens; this latter hypothesis is in good agreement with the increase of 5alpha-reduction activity observed at puberty or after treatment of young hypogonadal males. In addition, it is pointed out that a positive correlation is observed between the 5alpha-reductase activity present in each skin sample studied and the urinary 3alpha-androstanediol found for the same individual. This confirms our previous findings suggesting that the determination of urinary 3alpha-androstanediol might prove of clinical interest in the evaluation of the androgenic status in human subjects.

The possible role of 5 alpha-reduction of steroids in the sexual differentiation of guinea pigs was determined by treating pregnant guinea pigs with a 5 alpha-reductase activity (5 alpha RA) inhibitor (17 beta-N,N-diethylcarbamoyl-4-methyl-4-aza-5 alpha-androstan-3-one, 4MA, 10 mg/day) from day 30 to 55 of gestation. 5 alpha RA in fetal diencephalon tissue obtained from 4MA-treated mothers on day 55 of gestation was suppressed compared to that of control tissue. Four litters receiving 4MA were carried to term along with an equal number of litters receiving the vehicle alone. Males that received 4MA in utero (n = 6) had altered external genitalia, i.e., hypospadias and reduced anogenital distances, but their adult copulatory behavior did not differ from that of controls (n = 7). In order to evaluate treatment effects on the hypothalamic-pituitary axis, all animals were challenged with estradiol benzoate (EB, 10 micrograms in oil, s.c.) 2 weeks after gonadectomy. Serial plasma samples were obtained and analyzed for luteinizing hormone (LH) using an heterologous radioimmunoassay. Control females (n = 13) and 4MA-treated females (n = 5) released LH in surge quantities about 42 h after EB treatment. Plasma from 4MA-treated females differed from controls in that it contained greater overall quantities of LH (p less than 0.05) and greater amounts at the time of the LH surge (p less than 0.05). Regardless of treatment males did not respond to EB.(ABSTRACT TRUNCATED AT 250 WORDS)

Aspermatogenic seminiferous tubules were incubated in presence of labelled testosterone. This steroid is converted into androstenedione and 5 alpha-dihydro-testosterone. This finding favours the hypothesis which gives the Sertoli cell an important role in germ cell maturation.

In vitro study of testosterone (T) metabolism shows that formation of 17 beta-oestradiol (E2) in the preoptic area is higher in male doves exposed to behavioural stimuli from interacting pairs than in visually isolated males or males that have interacted with females. Formation of 5 alpha-dihydrotestosterone, 5 beta-dihydrotestosterone and 5 beta-androstane-3 alpha, 17 beta-diol was similar in all groups, indicating that exposure to stimuli from interacting pairs influences aromatization of T and not other pathways of androgen metabolism. The results suggest that sexual stimuli, which do not involve tactile contact between male and female, affect local E2 formation in the brain.

This study describes the recovery of various parameters of the pituitary-gonadal axis following five months of daily treatment of adult male rats with a potent LHRH (luteinizing hormone-releasing hormone) agonist. Two-month-old male rats were treated daily with either 250 ng or 1 microgram of [D-Ser(TBU)6, des-Gly-NH2(10)]LHRH ethylamide (LHRH-A) s.c. for five months. At the end of treatment, prostate weights were within normal limits and seminal vesicle weights were only slightly decreased. While normal values were found three months following cessation of treatment, it was observed, somewhat unexpectedly, that ventral prostate and seminal vesicle weights were increased by 66 and 54%, respectively, five months after cessation of treatment with the 1 microgram daily dose of LHRH-A. Immediately following the five-month treatment period with either dose of the LHRH agonist, basal testicular levels of pregnenolone, progesterone (P), 17-OH-progesterone (17-OH-P), androstenedione, testosterone, androstane-3 beta,17 beta-diol and androst-5-ene-3 beta, 17 beta-diol were decreased, while the concentrations of dihydrotestosterone (DHT), androstane-3 alpha, 17 beta-diol (3 alpha-diol) and 17 beta-estradiol were increased. Three months following cessation of treatment, all basal testicular steroid levels had returned to normal except pregnenolone, P, 17-OH-P and androstenedione, which were still reduced by 40 to 60%. Five months following cessation of treatment, on the other hand, basal levels of all testicular steroids were 40 to 200% increased in the animals having received either dose of the LHRH agonist. The testicular steroidogenic responsiveness was measured 2 hours following the subcutaneous administration of 10 micrograms oLH. Following five months of daily treatment with the LHRH agonist, the main findings are a decreased response of pregnenolone, P, 17-OH-P and androst-5-ene-3 beta, 17 beta-diol, and an increased DHT, 3 alpha-diol and androstane-3 beta, 17 beta-diol responsiveness. Three months post-treatment, on the other hand, particularly at the higher dose of LHRH agonist, there was an increased responsiveness of androstenedione, T, DHT and 3 alpha-diol, a finding which was maintained after two additional months of recovery. Degenerative changes were observed in most tubules following five months of LHRH-A treatment. While most tubules returned to normal five months later, some tubules still showed degenerative changes. Plasma LH measured by radioimmunoassay (RIA) was elevated after five months of treatment with the daily 1 microgram dose, but all other values were within normal limits.(ABSTRACT TRUNCATED AT 400 WORDS)

The growth and function of the human prostate is dependent upon a continuous supply of androgens, mainly 5 alpha-dihydrotestosterone, the 5 alpha-reduced metabolite of testosterone. Within the human prostate dihydrotestosterone is thought to be the intracellular mediator of androgen action. Although it is well documented that dihydrotestosterone is evenly distributed between the stromal and epithelial compartment of the prostate, the anatomical localization of dihydrotestosterone formation within the normal and hyperplastic prostate is still not established. To provide further insight into this problem we have measured, under conditions approximating the in vivo state, dihydrotestosterone formation in prostates obtained from 4 men with normal prostates and 36 men with benign prostatic hyperplasia. In addition to this we have performed histometric analysis of the cellular composition of the samples incubated, in order to correlate the morphological and the histochemical findings. Dihydrotestosterone is the major metabolite, and androstanediol and androstenedione were formed in smaller quantities. Under the given conditions metabolite formation from testosterone increased linearly for 60 minutes and the half maximum rate of dihydrotestosterone formation (Km) was observed at about 1.25 X 10(-6) M testosterone, a value similar to that reported for rat prostatic nuclei and human prostatic tissue. Dihydrotestosterone formation was higher in hyperplastic prostates than in the normal prostate. (Student's t test: p less than 0.05). The stroma in both the normal and hyperplastic tissue converts testosterone to dihydrotestosterone very actively. No significant relation was found between dihydrotestosterone formation and the per cent distribution of the stromal and epithelial compartment in any sample studied. In conclusion, our results are compatible first with the thesis that the rate of dihydrotestosterone formation is increased in the hyperplastic prostate and secondly with the concept that the rate of dihydrotestosterone formation is approximately the same in the epithelial and stromal compartments of the prostate.

Admixture of androgen-sensitive elements from normal or hyperplastic prostatic tissue interferes with biochemical studies of prostate cancer in its primary site. Heterogeneity of cancer tissues, varying in stromal and epithelial elements, also complicates interpretation of data relating to androgen metabolism. Accordingly, we have compared metastatic deposits composed of epithelial cancer cells to the primary biopsies of 4 patients in respect to uptake of 3H-testosterone and its conversion to 5-alpha-dihydrotestosterone during in vitro incubation. 3H-testosterone uptake was similar for both tissue sites but 3H-dihydrotestosterone formation was reduced by 76% in the metastases compared to primary tissues. This group was not large enough to show statistical significance, whereas a total of 11 such primary studies compared to 6 metastatic specimens was significant. When either primary or secondary tissue results were compared to 12 cases of benign prostatic hyperplasia similarly studied the differences were highly significant. These results demonstrate a major impairment in the formation of dihydrotestosterone by metastatic prostatic cancer and a similar but less evident alteration in the primary site. This abnormality in testosterone metabolism is of major importance in the attempt to obtain effective hormonal control of human prostatic cancer.

Growth characteristics, survival time, sex differences and hormonal effects, and various biochemical parameters were evaluated in a transplantable Furth/Wistar rat Wilms' tumor model. Survival time was dependent on site of tumor transplant and ranged from a mean of 28 days for intrarenal implantation to 44 days intramusculary. Maximum tumor weight (130 g) was obtained via subcutaneous implant. Lung metastasis was evident in the majority of animals with the exception of those receiving the tumor implant intraperitoneally. The levels of erythropoietin and serum calcium and phosphatase were comparable to control values whereas hematocrit levels declined. Tumor tissue arginase or total protein remained unchanged during tumor growth. In these same tissues DNA, content and 5-alpha-reductase activity significantly and progressively increased with concomitant tumor growths. Measurements of lactic dehydrogenase, alkaline phosphatase, and their isoenzymes indicated patterns of liver involvement which were not macroscopically evident. After 31 days of subcutaneous tumor transplant, male and female rats had tumors of comparable weights. Orchiectomy or estradiol treatment significantly reduced tumor weight in males. In female rats testosterone treatment significantly increased tumor weights. DNA concentration in tumor tissue was unaffected by treatment. Similiarly, although 5-alpha-reductase activity was higher in tumors from males, and arginase higher in females, these enzymes were not affected by surgical or hormonal treatment.

The sexual behaviour of male rats, castrated and testosterone-implanted, declined following induction of hyperprolactinaemia by domperidone. Treatment with oestradiol benzoate did not reverse this effect, and may have accentuated it. Oestradiol also amplified domperidone-induced hyperprolactinaemia. Testosterone or dihydrotestosterone (DHT) apparently delayed, but did not prevent, the gradual deterioration in sexual behaviour (prolonged ejaculation latencies) induced by domperidone, but this effect was not confirmed statistically. Adrenalectomy, followed by cortisol replacement, failed to prevent the behavioural effects of hyperprolactinaemia. No consistent changes in serum progesterone or corticosterone could be found in hyperprolactinaemic rats in which the adrenals had not been removed. In vitro formation of DHT from precursor testosterone was reduced in the amygdalae of hyperprolactinaemic rats, but not in the hypothalamus or caudal spinal cord. Oestradiol cytosol binding was unchanged in all brain areas, except for a small but significant increase in the anterior hypothalamus. These results do not support a role for altered adrenal activity in determining the effects of high levels of prolactin on sexual behaviour. There is evidence for an impaired formation of DHT in the brain, but this may account for only part of the behavioural changes observed. It is possible that the major effect of prolactin lies in neural systems directly responsive to it, rather than in altered steroid secretion or metabolism.

To characterize Leydig cell steroidogensis, we examined the metabolism of [3H]pregnenolone (3 beta-hydroxy-5-pregnen-20-one) to androgens in the presence and absence of human chorionic gonadotropin (hCG) as a function of culture duration. Approximately 20-30% of the (3H)pregnenolone was converted to testosterone (17 beta-hydroxy-4-androsten-3-one) by purified Leydig cells at 0, 3 and 5 days (d) of culture. Androstenedione (4-androstene-3,17-dione) and dihydrotestosterone (17 beta-hydroxy-5 alpha-androstan-3-one) were also produced while on day 5 of culture, significant amounts of progesterone (4-pregnene-3,20-dione) were isolated. The delta 5 intermediates, 17-hydroxypregnenolone (3 beta, 17-dihydroxy-5-pregnen-20-one) and dehydroepiandrosterone (3 beta-hydroxy-5-androsten-17-one), accounted for less than 1% of substrate conversion, indicating a clear preference for Leydig cells to metabolize (3H)pregnenolone via the delta 4 pathway. On day 0 of culture, unidentified metabolites considered of predominately polar steroids while on day 5 of culture, the unidentified metabolites consisted of predominately nonpolar steroids. In the presence of hCG, (3H-pregnenolone metabolism did not differ from basal on day 0 or 3 of culture. HCG increased the conversion of pregnenolone to progesterone and 17-hydroxyprogesterone (17-hydroxy-4-pregnene-3,20-dione) on 5d. This suggests that Leydig cells cultured for 5d have decreased C17-20 desmolase activity or that hCG acutely stimulates 3 beta-hydroxysteroid dehydrogenase and delta 5-delta 5 isomerase activities.

The splanchnic extraction and interconversion of testosterone and dihydrotestosterone (DHT) were studied in 7 healthy men (ages 29-46 years) undergoing cardiac catheterization. During a constant infusion of [1,2-3H]testosterone and [4-14C]DHT, the arterial and hepatic vein blood samples were taken and radioactive and non-radioactive testosterone and DHT were determined. Metabolic clearance rate (MCR), splanchnic extraction (SE), splanchnic clearance (SC), extrasplanchnic clearance (ESC), transfer constant in blood (T-DHT rhoBB) and transfer constant across the liver (T-DHT rhoSB) were calculated. The MCRT was 952 +- 172 (mean +- SD) 1/day and MCRDHT was 764 +/- 67 1/day in agreement with data from non-catheterized subjects. SET was 68.8 +/- 7.1% (mean +/- SD) and SEDHT was 37.6 +/- 5.9%. SET was significantly greater than SEDHT (P less than 0.001). The calculated SCT and ESCT were 638 +/- 112 (mean +/- SD) 1/day and 314 +/- 190 1/day, respectively. SCDHT and ESCDHT were 343 +/- 95 (mean +/-SD) 1/day and 421 +/-105 1/day, suggesting that a major fraction of testosterone is metabolized in the splanchnic organs and a higher fraction of DHT is metabolized in extrasplanchnic organs. In the interconversion study, overall conversion of testosterone to DHT in blood (T-DHT rhoBB) was 4.0 +/- 0.6% (mean +/- SD). No evidence for a net appearance of DHT by either mass or specific activity analysis in hepatic vein blood was observed in any infusion leading to the conclusion that the overall contribution of testosterone to circulating DHT from the liver (T-DHTrhoSB) was undetectable. This work indicates that conversion of testosterone to DHT occurs entirely in extrasplanchnic tissue in man.

The fetal guinea-pig testis synthetizes testosterone and reduces it in 5 alpha-dihydrotestosterone. The embryonic period which goes from the gonadla differentiation (on day 23) up to the beginning of the sex ducts differentiation (on day 29) is characterized in male guinea-pig by a very important testicular load in testosterone and DHT.

The subcellular distribution and functional characteristics of 5 alpha-reductase (3-oxo-5 alpha-steroid: NADP+ 4-ene-oxidoreductase, EC 1.3.1.22) from rat ventral prostate were studied and compared to the 5 alpha-reductase from female rat liver. Tissue fractionation retained main enzymic activity in the microsomal fraction of rat liver, while 5 alpha-reductase from rat prostate was localized in the nuclear membrane with a specific activity 160 times that of the initial homogenate. The purity of nuclear envelopes was checked by electron microscopy. Solubilization experiments indicated that the hepatic 5 alpha-reductase is attached to the endoplasmic reticulum as a peripheral protein, while the nuclear prostatic enzyme is an integral membrane protein. Incubation experiments with phospholipases suggested a decisive role of the surrounding phospholipids for the prostatic enzyme activity. To elucidate the characteristics of hydrogen transfer of the enzyme, the effect of flavins and different other cofactors on 5 alpha-reductase activity in isolated prostatic nuclei were studied. Our findings indicate that in rat ventral prostate the conversion of testosterone to 5 alpha-dihydrotestosterone proceeds by a direct hydrogen transfer from NADPH to testosterone. Concerning these parameters the behaviour of hepatic 5 alpha-reductase is absolutely different from the prostatic enzyme. The localization of 5 alpha-reductase within the nuclear envelope of rat ventral prostate as an integral membrane protein seems to be of physiological significance with regard to the action of androgens.