Differences in responsiveness to testosterone of penile reflexes and copulatory behavior of male rats

The display of penile reflexes and copulatory behavior appears to reflect the activity of two different underlying neuronal system, both of which are modulated by systemic testosterone (T) concentration. To indirectly compare the two systems, the responsiveness to T of penile reflexes and copulatory behavior was examined. In the first experiment castrated spinal male rats were given penile reflex tests while receiving replacement T through Silastic capsule implants filled with T (50 mm T). After capsule removal the number of penile erections and flips declined within 24 hr and gradually decreased for 12 days. Subjects were then reimplanted with new 50-mm T capsules. The number of penile flips and erections increased within 6 and 12 hr. respectively. This is a much more rapid response rate to T than has been established for copulatory behavior. In the second experiment castrated spinal male rats were tested for penile reflexes with a 50-mm T capsule, which was then replaced with a 10-, 5-, or 2-mm T or an empty capsule. The number of penile reflexes declined in a dose-response fashion. In the third experiment, castrated sexually experienced male rats were tested for copulatory behavior with two 25-mm T capsules which were then replaced with a 10 or 2-mm T or an empty capsule. Only males with empty capsules had decrements in copulatory behavior, revealing that a low level of T can maintain virtually normal sexual behavior despite a marked decline in penile reflex activity. The neuronal system underlying penile reflexes (spinal neurons) is apparently much more responsive to changes in T concentrations than the neuronal system underlying motivational and appetitive aspects of copulatory behavior (brain neurons).     

Serum estradiol, testosterone and dihydrotestosterone in male monkeys treated with testosterone propionate.

Serum estradiol (E2), testosterone (T) and dihydrotestosterone (DHT) were measured in juvenile (pre-pubertal) male rhesus monkeys injected with either 8 mg or 80 mg of testosterone propionate (TP). After one week, the three steroids were elevated and remained essentially unchanged for the duration of the study. There was little difference in serum E2 or DHT when comparing the two groups of steroid-treated monkeys. In contrast, T levels were consistently greater in the animals given the high dosage of TP.     

Sexual behavior and penile reflexes of neonatally castrated male rats treated in infancy with estrogen and dihydrotestosterone

Male rats castrated neonatally and treated with a combination of 0.5 μg estradiol benzoate (EB) plus 50μg dihydrotestosterone propionate (DHTP) for the next 14 days displayed normal sexual behavior when injected with testosterone propionate (TP) in adulthood. Neither EB nor DHTP alone had this developmental effect inasmuch as only 20–25% of the neonatal castrates treated with just 0.1, 0.5, or 10 μg EB, or 50 μg DHTP, displayed ejaculatory responses. The periodic application of mildly painful electric shock, which has been previously shown to markedly facilitate ejaculatory responding in normal male rats, failed to improve sexual performance in these latter subjects. This was true even of the castrates treated neonatally with DHTP which frequently intromitted. Castrates treated with EB or DHTP alone neonatally were subjected to spinal transection (after testing of sexual behavior) for examination of penile reflexes. Those treated with DHTP showed normal reflexes, characterized by numerous erections and flips, indicating the presumably nonaromatizable DHTP has developmental effects on penile reflexes similar to those of testosterone. Subjects treated with EB, including four animals that had ejaculated at least once, displayed very few, if any, erections on reflex tests and no flips. These results show that sometimes intromissive and ejaculatory patterns can occur even though the animal appears to have little or no capacity for penile reflexes.     

Effects of castration on androstenedione, testosterone and dihydrotestosterone plasma levels in adult male rats

In the present study we investigated the effects of castration on androstenedione (A), testosterone (T) and dihydrotestosterone (DHT) plasma levels in adult male rats 5 and 47 days after castration. In another group of 60-day-old castrated rats, the three steroids have been evaluated during testosterone propionate administration. Our data show that 5 days after orchiectomy all three steroids were significantly decreased (p less than 0.001) with respect to control values. 47 days after orchiectomy, T and DHT were also significantly decreased with respect to the control group. In both groups of orchiectomized rats the A/T ratio increased significantly with respect to controls. On the contrary, the T/DHT ratio sharply decreased. This suggests that DHT, in orchiectomized rats, could derive from precursors other than T. A negative correlation between A and the T/DHT ratio was observed 47 days after castration in adult animals and emphasized upon testosterone propionate administration. In the latter group, T was significantly lower while A is significantly augmented with respect to control values. Finally, the above-mentioned negative correlation indicates a possible prevalent role of A in contributing to the circulating levels of DHT in adult orchiectomized rats.     

Plasma testosterone and dihydrotestosterone in male rats during sexual maturation and following orchidectomy and experimental bilateral cryptorchidism.

Plasma concentration of testosterone and dihydrotestosterone (17beta-hydroxy-5alpha-androstan-3-one) were measured at frequent intervals (daily between day 20 and 40) in intact male rats from early pre-pubertal stage to full maturity. Effect of orchidectomy and experimental bilateral cryptorchidism on these two blood-born compounds at different stages of sexual maturation was also studied. A distinct spurt in testosterone level was noted on day 26, and in 3 days a leval of 2600pg/ml was reached, which was about 90% of the highest peak value (2940 pg/ml) seen at day 70. In the same animals dihydrotestosterone spurt was noted at the same time, but the peak value (163 pg/ml) was reached later (day 33). Within 4 days after orchidectomy plasma testosterone level showed no significant change in 20-day old animals, but dropped to 15% of the intact value in 40-day old ones, while remaining at a steady level of about 23% in older animals. Plasma dihydrotestosterone was undetectable after orchidectomy at each stage of sexual maturation. With bilateral cryptorchid animals plasma testosterone was higher than seen in the intact 20-day old control, but remained similar at other stages of sexual maturation. Plasma dihydrotesterone in the same animals was significantly less (p less 0.01) than those seen in the intact controls at all stages of sexual development except in 20-day old ones, where it was essentially similar.     

Differential effect of testosterone and dihydrotestosterone on the sexual behavior of prepuberally castrated male rabbits

The effect of testosterone propionate (Tp) and dihydrotestosterone propionate (DHTp) at doses of 1, 3 and 9 mg daily for 30 days on the copulatory behavior of prepuberally castrated male New Zealand white rabbits was studied. Tp was significantly more effective than DHTp in eliciting copulatory behavior at each dose level tested. Three milligrams Tp was the minimal dose required to elicit mounting consistently. DHTp at the high dose level (9 mg) only elicited sexual activity comparable to that observed with the low dose of Tp (1 mg). The results suggest that T does not need to be reduced to DHT to stimulate sexual behavior in the male rabbit.     

Plasma testosterone and sexual behavior following intracerebral implantation of testosterone propionate in the castrated male rat

Interpretation of behavioral and other effects of intracranial steroid implants depends on knowledge of the rate of release of the implanted hormones into the general circulation. Testosterone propionate implants (200 μg, pellets) in the median eminence and pituitary were found to result in circulating levels of testosterone (T) twice as high as those in the anterior hypothalamus-preoptic area (AHPOA), posterior hypothalamus (PH), and cortex (Ctx). Implants in all cranial areas examined resulted in plasma T levels in the lower range of circulating T found in normal rats for the first 24 hr postoperatively, decreasing thereafter and reaching very low levels by the end of 2 weeks. There were no significant differences in the plasma T levels in rats with implants in the AHPOA, PH, and Ctx, but AHPOA implants were slightly more effective in restoration of sexual behavior than PH implants, and both of these implants were considerably more effective than those in the cortex. There was no apparent correlation between behavioral responses and peripheral levels of T. The major conclusion of this study was that the effects of hypothalamic implants of T on male sexual behavior cannot be explained by the presence of T in the peripheral circulation.     

Developmental expression of penile reflexes and copulatory behavior in male rats.

First, pubertal development of the penile reflexes, e.g., erections, cups and flips in Wistar-Imamichi male rats was investigated following sheath retraction. Second, the penile reflexes and copulatory behavior in the above males were compared between 10 and 44 weeks of age. The penile reflexes in Wistar-Imamichi rats began to appear from day 26, and all males displayed full components of the reflexes on day 47. The occurrence rates and mean numbers of erections, cups and flips in aged adults were significantly low, compared with the young adults. Also, in the observation of copulatory behavior the occurrence rate of ejaculations, and mean numbers of intromissions and ejaculations in aged adults were significantly lower than that in young adults. These results of the present study may suggest that the decrease of copulatory behavior in male rats with age results from the dysfunction of the penile reflexes with age.     

Mounting behavior and lordosis behavior in castrated male rats treated with testosterone propionate, or with estradiol benzoate or dihydrotestosterone in combination with testosterone propinonate.

Treatment of prepuberally castrated male rats with testosterone propionate (TP, 50, 200, 500, or 1000 μg for 30 days) in adulthood stimulated the display of both mounting behavior and lordosis behavior. No correlation between mounting and lordosis behavior could be detected at any TP dose level. Treatment of prepuberally castrated male rats with either 1 μg estradiol benzoate (EB) or 500 μg dihydrotestosterone (DHT) for 60 days stimulated the display of mounting behavior in three of eight and four of eight rats, respectively. Treatment with 200 μg TP for the last 30 days of rats receiving either EB or DHT for 60 days resulted in an abrupt onset on mounting behavior as compared to rats treated with oil for 60 days. These results show additive effects of EB or DHT and TP upon mounting behavior by male rats and are interpreted as a support for the suggestion that testosterone to estrogen as well as testosterone to DHT conversion may be involved in the mechanism whereby testosterone activates the mounting behavior of castrated rats.     

Induction of male mating behavior in androgen-insensitive (tfm) and Normal (King-Holtzman) male rats: effect of testosterone propionate, estradiol benzoate, and dihydrotestosterone

Castrated androgen-insensitive rats exhibited mounting and intromission patterns in response to testosterone propionate (TP), estradiol benzoate (EB), or EB combined with dihydrotestosterone (DHT) treatment in adulthood. Treatment with DHT alone was ineffective in stimulating male mating behavior in the mutant rats. Since androgen-insensitive rats, like normal males, have the potential to show mounting behavior following hormone treatment in adulthood, the neural substrate underlying this behavior must be masculinized during development. The effectiveness of gonadal hormones in activating the entire copulatory sequence in castrated littermate males (King-Holtzman) was also examined. TP treatment induced mating behavior in the control rats. DHT also stimulated the complete copulatory pattern, although it was not as effective as TP. The administration of EB, however, did not induce ejaculation in control rats. These results do not support the hypothesis that the activation of male mating behavior by testosterone requires its metabolite estrogen (aromatization hypothesis).     

Effect of implanting bull calves with testosterone propionate, dihydrotestosterone propionate or oestradiol-17 beta prepubertally on the pituitary-testicular axis and on postpubertal social and sexual behaviour.

Twelve non-implanted crossbred bull calves served as controls and 30 crossbred bull calves (10/treatment) were implanted for 82 days, beginning at 34 days of age, to determine the influence of testosterone propionate (TP), dihydrotestosterone propionate (DHTP) and oestradiol-17 beta (E2) on prepubertal and pubertal pituitary-testicular function and on postpubertal social and sexual behaviour. Compared with control bulls, concentrations of serum luteinizing hormone (LH), follicle-stimulating hormone (FSH) and inhibin concentrations were suppressed (P less than 0.01) in all implanted bulls. Testosterone (T) concentration increased (P less than 0.001) in TP-implanted, but decreased (P less than 0.01) in DHTP and E2 bulls during the implant period. LH response to gonadotrophin-releasing hormone (GnRH) challenge during the implant period (2.5 months of age) was less (P less than 0.01) in TP, E2 and DHTP bulls than in controls. A small but significant T response to GnRH occurred in control bulls at 2.5 months of age. LH and T responses to GnRH challenge at 7 months of age (100 days after implant removal) was similar (P greater than 0.20) in control and implanted bulls. Steroid implants administered prepubertally had no effect (P greater than 0.10) on postpubertal social and sexual behaviours, including number of flehmen responses, abortive mounts, services and competitive order score. Body weight did not differ (P greater than 0.10) between treatment groups, but testis size was reduced (P less than 0.01) during the implant period and up to 10 months of age in treated bulls compared with controls. Testes remained smaller in E2-treated bulls up to the end of the study (23 months of age), but daily sperm production and epididymal weight did not differ (P greater than 0.10) between treatment groups at slaughter. Control bulls reached puberty earlier (P less than 0.01; 270 +/- 11 days of age) than did TP (302 +/- 11 days), DHTP (309 +/- 11 days) or E2 (327 +/- 11 days) bulls. Although puberty was delayed in all implant groups, there was no difference in scrotal circumference at puberty (average 28.4 +/- 0.4 cm) between treatment groups. Our findings indicate that TP, DHTP and E2 implants administered prepubertally result in acute suppression of serum LH, FSH and inhibin during the implant period and in post-implant suppression of testis size and delayed puberty in bulls. The lack of treatment effect on behaviour suggests that steroidal programming of sexual behaviour occurs before 1 month of age in bulls.     

Actions of esters of testosterone, dihydrotestosterone, or estradiol on sexual behavior in castrated male guinea pigs

Prepuberally castrated male guinea pigs were treated in adulthood with estradiol benzoate, testosterone propionate, dihydrotestosterone propionate or corn oil (vehicle control). Both corn oil and estradiol benzoate were ineffective in augmenting or inducing any aspect of adult male sexual behavior. Dihydrotestosterone propionate and testosterone propionate were both effective in establishing the complete male sexual behavior pattern, although they differed in the manner in which they affected specific components. For example, males treated with testosterone propionate showed more non-intromissive but not more intromissive mounts than males treated with dihydrotestosterone propionate. In addition, the average frequency of thrusts per intromission was greater for males treated with dihydrotestosterone propionate than for males treated with testosterone propionate.     

Percentage binding of testosterone and dihydrotestosterone and unbound testosterone and dihydrotestosterone in rabbit maternal and fetal plasma during sexual organogenesis.

The percentages of bound testosterone (17 beta-hydroxy-4-androsten-3-one; T) and dihydrotestosterone (17 beta-hydroxy-5 alpha-androstan-3-one; DHT) and their unbound concentrations were determined in pregnant rabbits and their fetuses from the 18th day of gestation to birth. T and DHT were also measured in fetal testes. In the testis, the total T/total DHT ratio, very high at 22 days (73.7 +/- 15.2), decreased until birth (6.7 +/- 0.8). In male fetuses the concentrations of total and unbound circulating T and DHT were always low and did not show any peak during sexual organogenesis. The percent binding of T (from 73.0 +/- 0.5 to 77.6 +/- 0.6) and DHT (from 76.5 to 83.7 +/- 1.1) in fetuses were similar in both sexes and significantly lower than those measured in mothers (T: from 87.2 +/- 0.6 to 91.6 +/- 0.9; DHT: from 87.3 +/- 0.9 to 93.8 +/- 0.9).     

Contribution of dihydrotestosterone to male sexual behaviour.

OBJECTIVE--To document the relative importance of endogenous sex steroids in modulating the frequency of orgasms, the dominant aspect of sexual behaviour in healthy eugonadal men. DESIGN--Measurement of adrenal and testicular sex steroids in a sample of army recruits and study of their relation to frequency of orgasms ascertained by questionnaire after potential confounding variables were controlled for. SETTING--Military campus and military hospital laboratories in Athens, Greece. SUBJECTS--92 consecutively enrolled healthy male recruits aged 18-22 years. MAIN OUTCOME MEASURES--Weekly number of orgasms. Serum concentrations of testosterone, dehydroepiandrosterone sulphate, dihydrotestosterone, oestradiol, oestrone, delta-4-androstenedione, and sex hormone binding globulin. RESULTS--Serum dihydrotestosterone concentration was the only independent hormonal predictor of the frequency of orgasms; an increase in concentration of 1.36 nmol/l (about 2 SD) corresponded to an average increase of one orgasm a week. CONCLUSIONS--Differences in concentrations of circulating dihydrotestosterone within the normal range may represent a major predictor of sexual activity in healthy young men.     

Testosterone,androstenedione and dihydrotestosterone: Effects on mating behavior of male rats

The relative effectiveness of testosterone, androstenedione, and dihydrotestosterone in maintaining mating behavior following castration of male rats was studied. In Experiment 1 testosterone, but not dihydrotestosterone, was found to maintain mating. In Experiment 2 testosterone and androstenedione were found to be equally effective in maintaining mating. Dihydrotestosterone failed to maintain mating and was no more effective than no treatment at all. Testosterone, androstenedione, and dihydrotestosterone significantly enhanced seminal vesicle and penis weight. In Experiment 3 castrated male rats were administered radiolabeled testosterone, androstenedione, or dihydrotestosterone. Radioactivity was found in hypothalamic and seminal vesicle samples indicating that these steroids can be accumulated by brain as well as peripheral androgen-sensitive tissues. It was concluded that the peripherally active steroid dihydrotestosterone probably plays no role in the maintenance of sexual behavior.