Intermale aggression and infanticide in aged C57BL/6J male mice: Behavioral deficits are not related to serum testosterone (T) levels and are not recovered by supplemental T

Healthy aged adul (24–26 months of age) and young adult (2–4 months of age) C57BL/6J male mice were assessed for intermale aggression, pup-killing behavior (infanticide), and circulating levels of testosterone (T). When compared to young adult male mice, aged adult males were highly variable in the exhibition of both androgen-dependent behaviors. Significant numbers of aged males exhibited deficits in aggression and pup-killing while other animals were as behaviorally active as their young male counterparts. Assessment of serum T showed that aging did not produce a reduction in levels of the steroid and individual variability in androgen-dependent behavior of aged males was not related to plasma levels of the hormone. When aged non-aggressive and non-killer males were exposed to supplemental T by way of subcutaneously implanted silastic capsules, circulating levels of the steroid were elevated but T-dependent behavior was not recovered. These findings, in combination with those previously reported for copulatory behavior, indicate that the deficits observed in the androgen-dependent behavior of aged male mice cannot be attributed to a breakdown in the production of testicular androgens. While neural refractoriness to T may account in part for deficits in androgen-dependent behavior of aged males, the variability that is observed in the reproductive behaviors of aged male rodents ultimately may be related to other sources of variation such as perinatal environment.     

Pup-killing in mice: the effects of gonadectomy and testosterone administration.

The administration of testosterone to intact and to gonadectomized adult female mice induced pup-killing whereas such treatment failed to induce killing in intact males and in males gonadectomized in adulthood. Testosterone treatment was effective in inducing killing when given to adult, neonatally gonadectomized, males. Also, gonadectomy of males performed prior to experience with young reduced the number of animals that subsequently killed. Gonadectomy performed after animals had killed pups did not eliminate killing behavior. Finally, practically all females that exhibited pup-killing in response to the administration of testosterone also displayed intraspecific aggression than killed pups.     

Genotypic influences on infanticide in mice: Environmental,situational and experiential determinants

Approximately 20–30% of adult, 60–70 day old, DBA/2J male mice exhibit infanticide toward standard stimulus 1–3 day old Rockland-Swiss Albino mouse pups, while 50–80% of adult C57BL/6J males display the behavior. Reciprocally-crossed hybrid males exhibit DBA-like levels of low pup-killing behavior and cross-fostered DBA and C57BL males retain their strain-typical phenotype. Also DBA males continue to exhibit low levels of infanticide and C57BL males display high levels of the behavior regardless of the length of pup exposure, or the sex, age, or strain of the stimulus newborn. Thus, strain differences in pup-killing behavior are not related to differences in the prenatal or postnatal maternal environment, or to situational or experiental factors. Inherent biological differences may be responsible for the strain differences in infanticidal behavior.     

Infanticide: Genetic,developmental and hormonal influences in mice

DBA/2J and androgen-deficient C57BL/6J mice were examined for their response to newborn (1–3 day old) Rockland Swiss (R-S) albino mouse pups. Significantly more 70–90 day old C57BL male mice killed young as compared to similarly aged DBA males (80% vs 30% respectively). Adult, 70–90 day old, female mice of both strains typically retrieved newborn young to the nest site. Adult castration significantly reduced infanticide in males of both strains while exposure to testosterone (T)-containing silastic capsules restored it. Treatment of adult ovariectomized female mice of both strains with T-containing capsules significantly elevated the exhibition of pup killing. When tested for infanticide at 25, 35, 45, 55 or 65 days of age, few males of either strain killed young at 25 days of age. However, beginning at 35 days of age, significantly more C57BL males killed young at every age as compared to DBA males. Moreover, C57BL males exhibited an earlier developmental onset of adult-like levels of infanticide than DBA males (45 vs 65 days of age respectively). Finally, older (4.5 to 7 months of age) DBA males exhibited levels of pup-killing identical to that of younger (2 months of age) C57BL males (70%). The findings are discussed in terms of their relationship to other sexually dimophic T-dependent masculine behaviors. The potential importance of infanticide for rodent reproductive strategies and population regulation is also considered.     

The role of androgens in the mediation of seasonal territorial aggression in male Siberian hamsters (Phodopus sungorus)

Testosterone (T) mediates aggression in a wide range of species. In some species, however, aggressive behavior persists or increases during the non-breeding season when T levels are relatively low. Animals that do not display a positive correlation between aggression and gonadal steroids suggest the need for further investigation of alternative neuroendocrine mechanisms mediating seasonal aggression. Siberian hamsters (Phodopus sungorus) are an ideal study system because they display increased territorial aggression during the non-breeding season which may be independent of circulating T levels. The goals of the present study were to: 1) explore the role of T in the aggression of reproductive males, and 2) test the hypothesis that the adrenal steroid dehydroepiandrosterone (DHEA) acts as an endocrine regulator of seasonal aggression. In Experiment 1, individuals were housed in long day (breeding) photoperiod and received castrations, exogenous T capsules or both manipulations. In Experiment 2, animals were housed in either long or short days (non-breeding) photoperiod and received DHEA or empty capsules. In both experiments, serum hormone levels and aggressive behavior were assessed. In Experiment 1, castration did not reduce aggression whereas exogenous T actually inhibited aggressive behavior. In Experiment 2, short-day individuals were more aggressive than long-day animals but DHEA treatment did not affect aggressive behavior, regardless of photoperiod. The present study supports the hypothesis that circulating gonadal steroids are not necessary to activate aggressive behavior in adult male hamsters. Further, seasonal changes in territorial aggression appear independent of circulating levels of DHEA in Siberian hamsters.     

Aggressive behaviors in adult SF-1 knockout mice that are not exposed to gonadal steroids during development

Sex hormones are a major factor responsible for the development of sex differences. Steroidogenic factor 1 (SF-1) is a key regulator of gonadal and adrenal development, and SF-1 knockout mice (SF-1 KO) are born without gonads and adrenal glands. Consequently, these mice are not exposed to gonadal sex steroids. SF-1 KO pups die shortly after birth due to adrenal deficiency. In the present study, SF-1 KO mice were rescued by neonatal corticosteroid injections followed by adrenal transplantations on day 7-8 postnatally. Control mice received corticosteroid injections and were gonadectomized prior to puberty. Mice were observed interacting with ovariectomized hormone primed females and gonad-intact males. In the absence of sex steroid replacement, adult SF-1 KO mice were significantly more aggressive than control mice in tests with stimulus females. After testosterone treatment, control males displayed significantly more aggression towards male intruders than control female mice, or male and female SF-1 KO mice, suggesting a developmental role of gonadal hormones in the expression of aggressive behavior and affirming SF-1 KO mice as a behavioral model to investigate affects of fetal gonad deficiency.     

Effects of pubertal anabolic-androgenic steroid (AAS) administration on reproductive and aggressive behaviors in male rats

Adolescence in human males is a hormonally sensitive period when many adult behaviors develop, including sexual and aggressive behaviors. Using a rat model, the authors examined the effects of three anabolic-androgenic steroids (AAS) during puberty: testosterone, nandrolone, and stanozolol. Copulation, vocalizations, scent-marking, and aggression were tested following AAS exposure. Relative to gonadally intact controls, rats injected with testosterone showed a significant increase in scent-marking and aggression in the opponent's home cage. Nandrolone had no effect. Stanozolol significantly inhibited all behaviors. Results suggest that depending on the chemical structure of the steroid, AAS exposure during puberty affects several androgen-dependent behaviors. Because adolescence in humans is a period of hormonal change, abuse of AAS, particularly stanozolol, during this time may disrupt the establishment of normal adult behavior patterns.     

Pup-killing behavior in mice: Suppression by early androgen exposure

Significantly more male mice castrated at birth killed newborn pups in response to adult testosterone propionate (TP) treatment than sham-operated controls. Female mice administered TP or estradiol benzoate (EB), but not testosterone (T), dihydrotestosterone (DHT), or dihydrotestosterone propionate (DHTP), on the day of birth killed significantly fewer young in response to adult TP than oil-treated controls. Neonatal MER-25 treatment prevented the suppressive effects of early TP exposure on T aroused pup-killing behavior. These data show that early androgen exposure attenuates the pup-killing producing properties of adult T and that aromatization of T to estrogen (E) during early life may mediate these effects.     

Effects of neonatal castration and testosterone on the rat's pup-killing behavior and activity

A male rat will generally kill young rat pups that are placed with him whereas a female generally will not. Another sex difference is that females are usually more active than males in the open-field test. This set of experiments was designed to evaluate the role of testosterone induced sexual differentiation in the development of these behaviors. The first experiment established that the frequency of the killing response is not diminished by castrating males in adulthood. The second experiment found that male rats which were neonatally castrated killed in fewer instances as compared to controls. These animals also were more active in the open field. Experiment 3 used males which had been handled or not disturbed in infancy to introduce an emotionality difference. Handling had no effect upon pup-killing behavior; again those castrated in infancy killed less often than controls. In Experiment 4 castrations at 2, 10 or 15 days of age were equally effective in stopping pup killing by males. In Experiment 5 females were androgenized at 2 days of age;this reduced their adult open-field activity but did not increase their pup-killing.     

Deletion of corticotropin-releasing factor binding protein selectively impairs maternal, but not intermale aggression

Corticotropin-releasing factor (CRF) binding protein (CRF-BP) is a secreted protein that acts to bind and limit the activity of the neuropeptides, CRF and urocortin (Ucn) 1. We previously selected for high maternal defense (protection of offspring) in mice and found CRF-BP to be elevated in the CNS of selected mice. We also previously determined that both CRF and Ucn 1 are potent inhibitors of offspring protection when administered centrally. Thus, elevated CRF-BP could promote defense by limiting endogenous actions of CRF or Ucn 1. To test this hypothesis, we crossed the deletion for CRF-BP into the mice selected for high maternal defense and evaluated offspring protection and other maternal behaviors. CRF-BP knockout (KO) mice exhibited significant deficits in maternal aggression relative to wild-type (WT) mice in three different measures. Other maternal features were almost identical between groups, including dam and pup weight, litter size, nursing time, and pup retrieval. Both groups performed similarly in a forced swim stress test and aggression in both groups was reduced following the swim test. Virgin KO female mice exhibited higher levels of anxiety-like behavior in terms of decreased time in the light portion of the light/dark box test. For males, no differences in light/dark box or swim test were found. However, increased anxiety-like behavior in male KO mice was identified in terms of contact and approach to a novel object both with and without previous exposure to the swim test. No differences in isolation induced resident intruder male aggression were found between groups. Together, these results indicate that loss of CRF-BP selectively impairs maternal, but not intermale aggression and that loss of the gene induces anxiety-like behavior in males and females, but there are sex differences in terms of how that anxiety is revealed.     

Effect of intrauterine position on sex differences in the gabaergic system and behavior of rats

In multiparous species such as the rat (in this case the albino Wistar strain), steroid influence during fetal growth is affected by the relative intrauterine position of male and female fetuses and is stronger when the potential effects of contiguity and caudal position are combined. The effect of intrauterine position on gonadal steroid levels in neonatal and adult animals was examined using radioimmunoassay techniques. Since the organizing effect of prenatal steroids may influence the postnatal GABA content, HPLC was used to determine the gabaergic content in several hypothalamic and limbic areas in the adult rat. The effects of intrauterine position on adaptive behavior were examined by recording exploratory behavior (using the corridor and hole board tests) and intraspecific aggression (induced by isolation). Female pups influenced by males during development produced more testosterone. In adult males, those that developed closer to the cervix (and with no influence from other fetuses) produced more testosterone and less estradiol. The same animals also produced more hypothalamic GABA and showed greater exploratory capacity. No significant differences were seen between any experimental groups with respect to aggression. These results show increased variability between males with respect to adult exploratory behavior, and in the neurochemical and endocrine systems involved, due to intrauterine position during development. The effect of this physiological phenomenon on the structure of rodent populations is discussed.     

Prenatal stress reduces intermale aggression in mice

Prenatal stress (heat and restraint) significantly reduced intermale aggression (percentage of animals fighting and the number of attacks and lunges) in Rockland-Swiss Albino mice. These data, in combination with previous reports showing deficits in male copulatory responses, suggest that a wide array of androgen-dependent social behaviors may be influenced by prenatal stress.     

Age-related deficits in brain androgen binding and metabolism, testosterone, and sexual behavior of male rats

Brain androgen binding and metabolism, serum testosterone (T), and sexual behavior were measured in old and young male Fischer 344 rats. After completion of sexual behavior tests, blood was collected for T assay and brains were removed for simultaneous measurements of cytosolic (ARc) and nuclear (ARn) androgen receptors and aromatase activity (AA) in the preoptic area (POA), hypothalamus (HYP) and amygdala (AMG). In Experiment 1, old and young intact males were examined. None of the old males ejaculated in any of the tests of sexual behavior whereas all of the young males ejaculated. The old males had lower levels of serum T, lower levels of ARn in the POA and HYP and lower levels of AA in the POA. The ARc levels of the old and young males did not differ. Experiment 2 was designed to determine if the deficits in brain androgen binding and metabolism were due to low levels of T. Old and young T-treated gonadectomized (GX-T) males and young intact (I) males were examined. T levels were comparable in the young and old GX-T males and were higher in each of these groups than in the young I males. In sexual behavior tests, all of the young but only 25% of the old GX-T males ejaculated. Although ARn levels in the old GX-T males were lower than in the young GX-T males, they were comparable to the young I male levels. No age-related differences in T induction of AA were observed.(ABSTRACT TRUNCATED AT 250 WORDS)     

Ethanol exposure during development reduces resident aggression and testosterone in rats

Ethanol exposure during development has been shown to alter social behaviors in people, but the range of deficits is not clear. Using an animal model of Fetal Alcohol Spectrum Disorders, inter-male aggression and testosterone levels were examined in adult rats. Rats were exposed to ethanol during the entire prenatal period and from postnatal day 2 through 10. Ethanol was administered via intragastric intubation. Two other groups consisted of a nontreated control and an intubated control group that was exposed to the administration procedures but not ethanol. Both offensive and defensive aggression were examined in experimental residents and intruders under three different housing conditions for the resident males: (1) with another male, (2) with a pregnant female, and (3) with a female and litter fathered by the experimental animal. When housed with a female and litter, ethanol-exposed rats displayed reduced offensive aggression compared to control groups under the same condition. Defensive aggression in the non-experimental intruders was reduced in this same condition. There were no differences in duration of non-aggressive social behaviors among the groups in any of the housing conditions. Testosterone levels were reduced in ethanol-exposed rats compared to controls. In summary, ethanol exposure over the combined prenatal and postnatal periods reduces aggressive behavior in a condition where aggressive behavior is normally seen. This reduction may be related to lower testosterone levels.     

Food availability affects behavior but not circulating gonadal hormones in maternal Belding's ground squirrels

We tested predictions of hypotheses suggesting that the steroid hormones, testosterone (T), progesterone (P), and estradiol (E2), contribute to the energetic regulation of behaviors associated with rearing young in free-living female Belding's ground squirrels (Spermophilus beldingi). We provisioned some female S. beldingi with food rich in fat and calories, and used unprovisioned females as controls. We observed the behavior of females throughout the reproductive cycle, and regularly collected blood samples to measure plasma hormone concentrations. Circulating concentrations of T, P, and E2 were similar in provisioned and unprovisioned females, as were temporal patterns of variation in these hormones. Peaks in rates of nest maintenance and aggressive behavior occurred during gestation and were associated with elevated concentrations of circulating T, P, and E2, raising the possibility that one or more of these hormones mediates behaviors that help females establish maternal nest sites and territories after mating. Temporal patterns of variation in behavior were similar among provisioned and unprovisioned females; however, rates of resting, vigilance, and aggression were higher among provisioned females, whereas unprovisioned females devoted significantly more time to feeding and locomotion. Thus, our data suggest that in maternal S. beldingi, gonadal steroids play a role in mediating behavior associated with raising offspring, but do not facilitate changes in rates of behavior associated with increased energy availability.     

Infanticide: accounting for genetic variation in mice

Infanticide, the killing of young, is one of a number of sexually-dimorphic traits in mice that is dependent upon androgen stimulation during perinatal life and during adulthood. Genotype also influences infanticide in that males of some strains of mice (C57BL/6J) exhibit high levels of this behavior while males of other strains (DBA/2J) seldom kill young. The experiments conducted here show that strain differences in pup killing behavior exhibited by males are not related to postweaning social factors nor are they due to differences in perinatal, pubertal, or adult levels of circulating hormones. These results, in combination with those previously reported, suggest that strain differences in the tendency of mice to kill young may instead depend upon the interaction of genotypic features such as prenatal hormone titers and/or sensitivity to these hormones, as well as on extra organismic factors such as intrauterine position. A model for understanding the manner in which genes and hormones may interact to influence infanticide and other hormone dependent sexually-dimorphic behaviors in mice is presented.     

Macrophages but not B cells from aged mice are defective in stimulating autoreactive T cells in vitro

In the present study the effect of aging on the capacity of Ia+ cells to stimulate autoreactive T cells in the syngeneic mixed lymphocyte reaction (SMLR) was investigated. Using young CD4+ T cells as responders, it was observed that unseparated whole spleen cells from aged mice had normal stimulatory activity comparable to that of young spleen cells. Interestingly, however, when purified splenic adherent cells (SAC) enriched for macrophages or splenic B cells were used as stimulators, aged SAC but not aged B cells were found to be defective in stimulating autoreactive T cells. This defect in aged SAC was not due to decreased expression of Ia antigens since the percentage of Ia+ SAC and density of Ia antigen expression was similar in both young and old mice. Also, the B cells from aged mice expressed normal levels of Ia antigens. Aged SAC, when mixed with young SAC could also actively suppress the normal SMLR. However, this suppression was not due to increased prostaglandin production but was found to be associated with interleukin-1 (IL-1) regulation, inasmuch as addition of exogenous IL-1 could completely reconstitute the defective stimulatory activity of aged SAC and also abolished the suppressor activity of the SAC. Aged mice also demonstrated an intrinsic defect in the CD4+ T cells responding in the SMLR. Together, our studies on the SMLR demonstrate an age-related defect in responder autoreactive T cells and in stimulator splenic macrophages but not in the stimulatory activity of B cells.     

Skin conductance fear conditioning impairments and aggression: A longitudinal study

Autonomic fear conditioning deficits have been linked to child aggression and adult criminal behavior. However, it is unknown if fear conditioning deficits are specific to certain subtypes of aggression, and longitudinal research is rare. In the current study, reactive and proactive aggression were assessed in a sample of males and females when aged 10, 12, 15, and 18 years old. Skin conductance fear conditioning data were collected when they were 18 years old. Individuals who were persistently high on proactive aggression measures had significantly poorer conditioned responses at 18 years old when compared to others. This association was not found for reactive aggression. Consistent with prior literature, findings suggest that persistent antisocial individuals have unique neurobiological characteristics and that poor autonomic fear conditioning is associated with the presence of increased instrumental aggressive behavior.     

The role of 5alpha-reductase activity in sexual behaviors of the green anole lizard

Both testosterone (T) and its metabolite, 5alpha-dihydrotestosterone (DHT), can facilitate male sexual behavior in the lizard Anolis carolinensis. The present study addresses the role of DHT synthesis in regulating male sexual behavior by inhibiting 5alpha-reductase, the enzyme that converts T into DHT. In two separate experiments (one replacement and one maintenance paradigm), breeding adult males were castrated and implanted with capsules of T, DHT, or a control capsule (blank, BL). The animals were then injected with the 5alpha-reductase inhibitor, FCE, or with steroid suspending vehicle (SSV) as a control. Both experiments produced similar results. Overall, T was most effective in eliciting courtship and copulatory behaviors above control levels. In both experiments, treatment with FCE attenuated the T-induced effects on courtship behavior, whereas the inhibition of 5alpha-reductase activity resulted in modest and inconsistent effects on the latency to intromission and the proportion of copulating males. DHT treatment did not significantly increase courtship or copulatory behaviors above control levels. These results suggest that (a) 5alpha-reductase activity is necessary but that DHT alone is not sufficient for stimulating courtship in male A. carolinensis; and (b) courtship behavior is more sensitive than copulatory behavior to the activity of the androgen metabolizing enzyme.     

Testosterone,puberty, and the pattern of male aggression in Syrian hamsters

The interaction between testosterone and pubertal development on aggression in the male Syrian hamster (Mesocricetus auratus) was examined in two experiments. First, gonad-intact prepubertal and adult male hamsters were tested for aggression using the resident-intruder paradigm with an age- and weight-matched intruder. Prepubertal males engaged in a greater number of attacks and had longer attack durations than adults. In the second experiment, prepubertal and adult males were castrated and treated with either a 0-, 2.5-, or 5-mg pellet of testosterone, and their aggression was assessed using the resident-intruder paradigm. Prepubertal males again showed shorter attack latencies and greater attack durations compared to adult males across all levels of testosterone. Testosterone treatment did not significantly affect these behaviors before or after pubertal development. We conclude that (a) pubertal development is associated with a decline in aggression in the male Syrian hamster, and (b) a developmental shift in behavioral sensitivity or responsiveness to testosterone does not underlie the pubertal decrease in aggression.