Role of corticosterone in the control of post-shock fighting behaviour in male laboratory mice

Two experiments were conducted to examine the role of corticosterone in controlling fighting subsequent to electric foot-shock in male mice. It was found that (1) treatment with corticosterone restores post-shock fighting in adrenalectomized males but not in castrated-adrenalectomized males; (2) raising cortieosterone levels induces post-shock fighting in castrated-adrenalectomized males treated with testosterone but not in castrated-adrenalectomized controls treated with a placebo; and (3) preventing corticosterone responses to shock occludes the increasing in fighting which normally follow shock in intact males. These findings suggest that (1) corticosterone plays an important role in the control of post-shock fighting in mice; (2) this importance of corticosterone is dependent on the presence of circulating testosterone, or other androgens; and (3) corticosterone responses to the experience of shock may be involved in the mediation of the facilitatory effects of this experience on subsequent fighting behaviour.     

Role of the pituitary-adrenocortical axis in the control of agonistic behaviour after single and repeated footshock in castrated male mice

Two experiments were conducted to examine the role of the pituitary-adrenocortical axis in the mediation of the effects of single and repeated electric footshock on subsequent agonistic responding in castrated male mice. It was found that (1) preventing corticosterone responses to shock occludes the facilitatory effects of single shock on both aggressive and submissive behaviour and occludes the additional increases in submissive behaviour which normally occur after repeated shock, and (2) blocking pituitary release of ACTH by dexamethasone treatment restores aggressive behaviour after repeated shock, independently of the initial levels of corticosterone and testosterone. These findings suggest that (1) increases in aggressive and submissive behaviour in response to single shock depend on shock-induced increases in corticosterone levels; (2) further increases in submissive behaviour in response to repeated shock require further increases in the corticosterone levels; and (3) inhibition of aggressive behaviour after shock repetition appears to depend on increases in ACTH levels, and occurs independently of the initial corticosterone and testosterone levels.     

Effect of sex hormones on the growth and multiplication of tetrathyridia of Mesocestoides corti (Cestoda: Cyclophyllidea) in mice

The number of tetrathyridia in the peritoneal cavities of mice increases exponentially with time. Thirty days post infection more larvae are in the cavities than in the livers. After that the increase of intraperitoneal populations continues, whereas the number of tetrathyridia in the livers remains more or less constant.Exogenous testosterone propionate, 10 $\text{μg}\text{g}$, twice a week, for 5 weeks, increases significantly the total volumes of tetrathyridial populations in the peritoneal cavities, whereas oestradiol benzoate, 5 $\text{μg}\text{g}$ or 10 $\text{μg}\text{g}$, also for 5 weeks, accelerates the rate of growth and multiplication of coelomic tetrathyridia to a much lesser extent, but increases significantly the infection of the livers.     

Individual differences in the attack behavior of male mice: a function of attack stimulus and hormonal state

Male CFW mice were tested for fighting behavior directed against olfactory bulbectomized male mice and against lactating female mice. Some males were tested with each stimulus type before and after castration. Some males were tested first following castration and then after testosterone treatment. All gonadally intact males attacked bulbectomized males and 25% attacked lactating females. After castration 81% attacked males on at least one occasion and 62% began to attack lactating females, although individual differences in the pattern of post-castration behavior were large. Individual differences in attack behavior were also large in males whose first tests followed castration. Of these, 60% attacked both stimuli. Following testosterone treatment, attack against males increased while attack against females was inhibited. Hormonal stimulation reduced individual differences in behavior and increased males' discrimination between the two types of stimuli.     

Puberty in the male pony: Plasma gonadotropin concentrations and the effects of castration

Ten male ponies were studied from 17 December 1978 through 9 August 1979. Six of the colts were born the previous spring (1978) and four were born during the previous summer. Three of the spring-born colts had been castrated at 4 months of age. Based on the presence of spermatozoa in the epididymis, all spring-born colts ($\text{3}\text{3}$), but only one summer-born colt ($\text{1}\text{4}$) had reached puberty by the end of the project (August). Spermatogenesis was significantly more advanced in the spring-born colts than in the summer-born colts.Concentrations of FSH and LH in the intact males remained constant from December through August, and levels were significantly lower than for the long-term castrated males throughout this period. In the long-term castrates, concentrations of FSH and LH increased from late winter and early spring to the highest levels during late spring and summer.On 9 August, the three spring-born colts (approximately 16 months of age) were castrated. The four summer-born colts (approximately 12–13 months old) were randomly assigned to either castrate (n=2) or hemicastrate (n=2) groups, and surgery was done on all colts on the same day. Both gonadotropins increased following castration in spring-born males. Concentrations of FSH and LH did not change following hemicastration.     

Postcastration retention of sexual behaviour in the male BDF1 mouse: The role of experience

Male BDF₁ mice (the F₁ progeny of a cross between C57BL/6 females with DBA/2 males) show a remarkable retention of sexual behaviour following castration. Two experiments were conducted to describe in detail the postcastration copulatory performance of the BDF₁ male mouse and to determine to what extent such performance is influenced by experience prior to castration. Experiment I found that castration leads to significant increases in the number of mounts and intromissions needed to reach ejaculation, and to a significant increase in ejaculation latency. Experiment II found that although precastrational sexual experience is not essential for the performance of the ejaculatory reflex after castration, it does influence the frequency of its occurrence. Furthermore, type of postweaning social experience influenced the display of ejaculatory behaviour by non-experienced castrates, as those with female social experience were superior to those with social experience with males or no social experience. The interactions of experience, hormones and genotype in the control of sexual behaviour in the BDF₁ male are discussed.     

Chronic treatment with valium (diazepam) fails to affect the reproductive system of the male rat

Male rats treated chronically with high doses of Valium (50mg/ Kg/day; 10 days) failed to exhibit changes in their reproductive system. Testicular and prostate weights, serum testosterone (T) and LH were unaffected. Testes and pituitary tissue stimulated $\text{in}$$\text{vitro}$ with LH and GnRH, respectively, released normal amounts of T, LH and FSH. Brain benzodiazepine receptors were slightly but significantly elevated by Valium treatment as well as by castration. We conclude that the male reproductive system is resistant to chronic Valium treatment even though the brain levels of benzodiazepine receptors are not.     

Dominance and aggression in social groups of male and female rats

Two experiments were performed to examine aggression and dominance in domestic male and female $\text{Rattus norvegicus}$ living in small mixed-sex (3 males and 3 females) groups. Experiment 1 examined the development of aggression in females. A single female (alpha) within each of the six colonies tested showed the preponderance of attacks on male intruders placed into the home-cage when male colony residents were absent. Over 12 weeks of intruder-aggression training female alphas showed only a mild nonsignificant elevation of aggressive behavior. A comparison of aggression of male and female colony alphas tested with opponents of each sex revealed that aggression was mainly directed at like-sex opponents, and that female attack was more defensive in character than male attack regardless of opponent sex. The highest intensity of aggression occurred when male alphas confronted male intruders. Although intruders never showed offense toward male residents, 61% of intruding males showed offense in response to attack by females.Experiment 2 investigated the relationship between aggressive dominance and competitive measures of dominance within each of 10 mixed-sex colonies. Alpha stat s of male and female colony residents did not reliably predict priority of access to food or water in tests of direct resource competition with like-sex colony members. When colony males were simultaneously tested for copulation, the copulatory behavior of alpha males was significantly greater than that of other colony males. Results are discussed in relation to the role of aggression in the reproductive strategy of male and female $\text{Rattus norvegicus}$.     

A novel nuclear protein in rat ventral prostate androgen-dependent and age-related change

A novel protein was found in the nuclei of rat ventral prostate. This protein has a molecular weight of about 21 kDa as measured by SDS-polyacrylamide gel electrophoresis. It showed a characteristic change between 3 and 84 weeks after birth in close association with the level of testosterone in the blood. After castration, the level of the 21-kDa protein decreased to $\text{1}\text{60}$ of normal in 7 days, but on daily injection of testosterone the level was restored to normal in 8 days and to twice the normal level in 14 days. Unlike H₁ and H₁⁰ histone and high mobility group proteins, the 21-kDa protein was not extracted with 5% HClO₄, but was partially extracted with 0.35 M NaCl. The 21-kDa protein was not found in kidney, liver, or brain, suggesting that it is specific to the ventral prostate.     

Naloxone effects on a nociceptive response of hypophysectomized and adrenalectomized mice

Naloxone (1 and 3 mg/kg) or saline was administered to adrenalectomized, sham-adrenalectomized, hypophysectomized, and sham-hypophysectomized mice prior to placing them on a 55° C hot plate. Naloxone reduced the jump latency of the adrenalectomized, sham-adrenalectomized, and sham-hypophysectomized mice, but did not reduce the jump latency of hypophysectomized mice. Hypophysectomy and adrenalectomy $\text{per se}$ did not significantly affect the jump latency. These results indicate that the pituitary is necessary for naloxone to reduce the escape latency of mice on the hot plate. The possibility of a hyperalgesic factor is suggested.     

Stimulation of leydig cell testoterone secretion in vitro and in vivo in hypophysectomized rats by an agonist of luteinizing hormone releasing hormone

Injection of a luteinizing hormone-releasing hormone (LHRH) agonist into 55-day-old male rats which had been hypophysectomized 3 days earlier resulted in a 10- to 30-fold increase in the levels of testosterone in serum and testicular interstitial fluid (IF) in the 4h following injection. The levels achieved were within or above the normal range for intact untreated rats of this age. In similar animals, injection of LHRH agonist also enhanced the serum testosterone response to injected hCG at $\text{1}\text{1}\text{2}\text{h}$, but not at later times after injection, and by 24h reduced IF levels of testosterone suggested that LHRH agonist had begun to inhibit stimulation by hCG. $\text{In vitro}$, dispersed Leydig cells from untreated hypophysectomized rats showed a 2-fold increase in testosterone responsiveness to LHRH agonist when compared to cells from intact rats, and this change was associated with an 80% increase in the number of Leydig cell LHRH-receptors.     

Differential responsiveness to replacement therapy of pre- and postpubertally castrated mice with respect to intermale aggression

Male mice castrated before Day 6 of postnatal life differ from adult castrates in that they do not exhibit normal patterns of intermale aggression following replacement therapy as adults. Two experiments sought to determine the effect castration after Day 6, but before puberty, would have on this response to adult replacement therapy. It was found that adult castrates showed some increases in fighting as early as six hr after a subcutaneous (s.c.) injection of 2 mg testosterone propionate (TP). At 30 hr after treatment 14/15 mice fought, while at 72 hr 15/15 fought and the number of fights in 10 min was significantly higher than at 30 hr. In the second experiment, the response to adult treatment with 2 mg TP (s.c.) in mice castrated in Day 10 or Day 50 was compared. Different groups were tested at 16, 40, 64, and 88 hr after TP treatment. The latency to respond to TP was significantly less and the level of fighting obtained was significantly greater in Day 50 castrates than in Day 10 castrates at each time of testing. The proportion of Day 10 (14/15) and Day 50 (15/15) castrates fighting was equal by 88 hr, while the mean fighting frequency was significantly lower in Day 10 castrates (M = 4.93 ± 4.78) than in Day 50 castrates (M = 8.06 ± 1.63). Day 50 castrates fought significantly more often than controls at 40 hr, whereas the level of fighting attained by Day 10 castrates was not significant until 64 hr. These results suggest that even after the organizational period for aggression, testosterone is necessary for maintenance and/or preparation of the substrate essential to the elicitation of aggression.     

Polyamine metabolism in the kidneys of castrated and testosterone-treated mice after administration of methylglyoxal bis(guanylhydrazone)

The effects of methylglyoxal bis(guanylhydrazone) on $\text{S-}\text{adenosyl}\text{-}\text{l}\text{-}\text{methionine}$ decarboxylase (EC 4.1.1.50) activity were studied in the mouse kidney stimulated to growth by testosterone administration. The drug was found to be a potent inhibitor of the enzyme in vitro. Administration of methylglyoxal bis(guanylhydrazone) in vivo resulted in a transient inhibition followed by a strong enhancement of the enzyme activity. Dialysis of the kidney extract, to remove remaining methylglyoxal bis(guanylhydrazone), revealed a great and rapid increase in the activity of $\text{S-}\text{adenosyl}\text{-}\text{l}\text{-}\text{methionine}$ decarboxylase. Injections of testosterone to castrated mice resulted in a marked increase in kidney weight and an accumulation of renal putrescine, spermidine and spermine. These effects of testosterone could not be blocked by simultaneous injections of methylglyoxal bis(guanylhydrazone). It appears that due to secondary effects by which the inhibition of methylglyoxal bis(guanylhydrazone) on $\text{S-}\text{adenosyl}\text{-}\text{l}\text{-}\text{methionine}$ decarboxylase activity is circumvented the inhibitor seems to be of uncertain value in attempts to decrease selectively the in vivo levels of polyamines.     

Genotype and retention of the ejaculatory reflex in castrated male mice

Genotype proves to be an important factor affecting the retention of sexual responsiveness following castration. Male F₁ hybrids between C57BL/6 females and DBA/2 males (B6D2F₁) show very prolonged retention. Experiment 1 used a diallelic design using C57BL/6, DBA/2 and BALB/c strains and showed that heterosis per se is not an adequate explanation of the superior retention of the B6D2F₁ genotype. Experiment 2 showed that neonatal injections of 100 μg TP to B6D2F₁ male mice did not result in better retention of the ejaculatory reflex after castration in adulthood. B6D2F₁ males from both experiments who exhibited the ejaculatory reflex for many months after castration went through a ‘difficult period’ in the first 10 to 12 postcastration weeks. During this time, ejaculation latency increased fourfold, and then returned to the original level. On the other hand, the proportion of males reaching the ejaculatory threshold during this period first declined then increased.     

In vitro androgen metabolism in mouse kidney: High 3-keto-reductase (3α-hydroxysteroid dehydrogenase) activity relative to 5α-reductase

The $\text{in vitro}$ metabolism of testosterone and dihydrotestosterone was studied in slices and cell fractions of mouse kidney. When testosterone was used as substrate, very little metabolism to dihydrotestosterone occurred suggesting very low 5α-reductase activity. When dihydrotestosterone was substrate, it was rapidly converted to 5α-androstane-3α, 17β-diol by a potent 3-keto-reductase. Ninety-five percent of this latter enzyme is located in cytosol and it requires NADPH as cofactor. The 3-keto-reductase may exist in two molecular forms which can be separated by polyacrylamide gel electrophoresis. Form A and B have mean molecular radii which correspond to molecular weights of 38, 700 and 28, 700, respectively. Sufficient 3-keto-reductase activity is present in cytosol at 0°C to reduce physiological concentrations (2×10^?9 M) of dihydrotestosterone without the addition of cofactor. 3-Keto-reductase activity is higher in intact male than in castrate male or female mice and increases with androgen treatment.From these studies we conclude: (a) The virtual absence of 5α-reductase in mouse kidney is consistent with the thesis that testosterone rather than dihydrotestosterone may be the intracellular androgen in this organ. (b) Kinetic studies which depend upon the $\text{in vitro}$ uptake and retention of dihydrotestosterone by receptor proteins may be difficult to interpret due to rapid metabolism of ligand.     

In vitro androgen metabolism in mouse kidney: high 3-keto-reductase (3alpha-hydroxysteroid dehydrogenase) activity relative to 5alpha-reductase

The $\text{in vitro}$ metabolism of testosterone and dihydrotestosterone was studied in slices and cell fractions of mouse kidney. When testosterone was used as substrate, very little metabolism to dihydrotestosterone occurred suggesting very low 5α-reductase activity. When dihydrotestosterone was substrate, it was rapidly converted to 5α-androstane-3α, 17β-diol by a potent 3-keto-reductase. Ninety-five percent of this latter enzyme is located in cytosol and it requires NADPH as cofactor. The 3-keto-reductase may exist in two molecular forms which can be separated by polyacrylamide gel electrophoresis. Form A and B have mean molecular radii which correspond to molecular weights of 38,700 and 28,700, respectively. Sufficient 3-keto-reductase activity is present in cytosol at 0°C to reduce physiological concentrations (2×10^?9 M) of dihydrotestosterone without the addition of cofactor. 3-Keto-reductase activity is higher in intact male than in castrate male or female mice and increases with androgen treatment.From these studies we conclude: (a) The virtual absence of 5α-reductase in mouse kidney is consistent with the thesis that testosterone rather than dihydrotestosterone may be the intracellular androgen in this organ. (b) Kinetic studies which depend upon the $\text{in vitro}$ uptake and retention of dihydrotestosterone by receptor proteins may be difficult to interpret due to rapid metabolism of ligand.     

Correlated effects of the Y-chromosome of mice on developmental changes in testosterone levels and intermale aggression.

Serum testosterone (T) was studied developmentally in DBA/1/Bg and C57BL/10/Bg inbred mice, as well as in their reciprocal F1 hybrids. Testosterone determinations were made using a radioimmunoassay. At 35 and 40 days $\text{post partum}$, DBA/1/Bg mice had higher levels of T than C57BL/10/Bg males. Comparison of the regression coefficients for the serum T values over days 30, 35, 40, and 50 also indicated statistical differences in the two curves of developmental changes in T. The pubertal rise in B10D1F1 but not D1B10F1 males was found to be steeper in slope over days 30, 35, 40, and 50 than that of C57BL/10/Bg males. These data suggest that during the pubertal period there may be a more rapid increase in serum T titer associated with the DBA/1 Y-chromosome. However, the developmental curves of T for the two reciprocal hybrids were not statistically different from each other or from that of the DBA/1/Bg males. These findings are suggestive of a Y-chromosome effect on developmental changes in T and of a genetic correlation with intermale aggression.     

B-ETA-Endorphin immunoreactivity in rat and human blood: radioimmunoassay, comparative levels and physiological alterations.

Antiserum against human β-Endorphin (β_hEP) has been obtained from rabbit. The antiserum, diluted $\text{1}\text{1500}$ bound I¹²⁵ β_h-EP, demonstrating an effective range from 10pM to 10nM. The sensitivity of the assay is 2–3 fmoles. This antibody exhibits 10–15% cross-reactivity with human β-Lipotropin (β_h-LPH). β-EP-like immunoreactivity in rat blood has been detected in unextracted samples when compared to blood from hypophysectomized rats. The whole assay and calibration curves are carried out in plasma from hypophysectomized animals. β-EP-like immunoreactivity can be detected in normal rat plasma (75 ± 15 fmole/ml), and exhibits substantial increases with adrenalectomy (287 ± 32 fmoles/ml). In contrast, samples from five healthy normal human males gave values near the limits of detection of the assay (12 fmoles ± 3.9 per ml of plasma). Such values may be due to cross-reactivity of the antiserum with β_h-LPH or other circulating hormones. In contrast, patients with elevated ACTH production and normal pregnant humans exhibit significantly elevated levels of β-EP immunoreactivity in plasma.     

Exposure of embryos to an aromatization inhibitor increases copulatory behaviour of male quail

This experiment examined the possibility that endogenous embryonic androgen contributes to sexual differentiation of behaviour in male or female quail ($\text{Coturnix}$$\text{coturnix}$$\text{japonica}$), and that it does so via aromatization (conversion to oestrogen). Eggs were injected on day 9 of incubation with oil or ATD (an aromatization inhibitor). As adults, males and females were exposed to short days, injected with testosterone propionate, tested for male-typical behaviour, then injected with oestradiol benzoate and tested for female-typical receptivity. ATD increased the level of male-typical copulatory behaviour in males. Male-typical behaviour in females was not affected, nor was female-typical behaviour in either sex. Thus normal male quail are actually slightly demasculinized by their own androgen during embryonic development, and this process is mediated by aromatization.