Differences in the capacity of male odours to affect investigatory behaviour and different urinary marking patterns in two strains of mice,selectively bred for high and low aggressiveness.

The investigatory behaviour of male and female mice on sawdust soiled by male mice with different levels of aggressiveness was studied in two experiments. To investigate whether high and low aggressive males show different urinary marking patterns, a third experiment was set up. The strains TA(Turku Aggressive) and TNA (Turku Nonaggressive) have been developed by selective breeding. The TA-soiled bedding discouraged investigation by male mice, while females avoided TNA-soiled areas. Also the urine marking patterns differed between the high aggressive TA and low aggressive TNA males. The results indicate that the urine marking behaviour and the odour communication system in the TA and TNA males correlate with their hereditarily determined disposition for aggressive behaviour.     

Urine marking and social dominance in male house mice (Mus musculus domesticus)

House mice use urine marking for a variety of forms of social communication. Urine marking varies with dominance status; socially dominant male house mice urine mark more than those that are socially subordinate. Experiment I was designed to confirm this previous finding. Experiment II was designed to test whether urine marking, measured prior to testing males for aggression, could be used to predict social dominance. Mice were tested for urine marking in 20 cmx40 cm rectangular cages with filter paper below the wire mesh bottom of the cage. In Experiment I, groups of four males were tested in a round robin design to assess social dominance and were then placed individually in urine marking cages. Social dominance was a significant predictor of the number of 1 cm squares that contained urine marks, both with regard to interior squares and for perimeter squares in the test cage. In Experiment II, groups of four males were first tested individually in urine marking cages and then used for round robin aggressive encounters to assess social dominance. The number of interior squares with urine marks, and, to a lesser extent, the number of perimeter squares with urine marks, were both significant predictors of aggression scores and social dominance status. Being able to judge social dominance without having the mice encounter each other could be a valuable tool for future work; confounding effects on such parameters as hormone levels could be avoided while obtaining an estimate of male social dominance status.     

The hormonal control of scent marking and precopulatory behavior in male gray short-tailed opossums (Monodelphis domestica)

Little is known about the hormonal control of behavior in marsupials. In the present study, the effects of castration and of testosterone or estradiol replacement therapy on scent marking and precopulatory behavior in male gray short-tailed oppossums (Monodelphis domestica) were examined. It was found that castration resulted in decreases in chest and flank/hip marking displayed by male gray opossums. Testosterone but not estradiol stimulated chest marking in castrates. Males treated with either estradiol or testosterone displayed more flank/hip marking than control males. Highest levels of female aggression toward males were seen when the males had received testosterone treatment. These findings are discussed with respect to similarities and differences between marsupials and eutherians in the neural metabolism of testosterone and the hormonal control of scent marking behavior.     

Individual odours and mate recognition in the prairie vole,Microtus ochrogaster

A habituation-discrimination technique was used to demonstrate that male and female prairie voles can discriminate individual differences in the odours of soiled shavings and urine from male and female conspecifics. A second experiment, employing a Y-maze, showed that females significantly preferred the odours of their mate over those of either another mated male or an unmated male. Males preferred their mate's odours to those of other mated females but showed no significant preference between the odours of their mate and those of a virgin female. A third experiment demonstrated that, over a 10-h period, females built nests and/or stayed preferentially on the side of a Y-maze containing their mate's odours. Likewise, males preferentially built nests in their mate's side compared to the side containing odours from a virgin female. However, although the same trend was present when mate odours were paired with odours from another mated female, the preference was not statistically significant. Taken as a whole, these results indicate that mate recognition may be an important of individually distinctive odours in this species.     

The development of aggressive and marking behavior in intact and castrated male hamsters.

In pairs consisting of two intact or two castrated male hamsters, the level of aggressive and flank-marking behavior increased with age. Dominance relationships were established early and remained stable. Castration prevented the appearance of the flank gland but did not alter the development of aggression, dominance, and marking. After the formation of intact-castrate pairs, the level of performance of the intacts was significantly higher than the castrates only for marking behavior. In a second experiment involving 49 intact-castrate pairs, there were no differences between intacts and castrates for any behavioral variables measured. Although testicular integrity may influence agonistic behavior in certain situations, these data indicate that it must be only one of several influences in determining the occurrence and outcome of an agonistic encounter.     

Gonadal hormones and conspecific marking in male rats

Urine deposited by a rat on a conspecific was quantified with injections of sodium fluorescein, a substance that changes the color of urine. The hypothesis examined in experiment 1 was that marking the environment and a conspecific would be similarly androgen-sensitive behaviors during each of three stages--before castration, after castration, and with restorative therapy with testosterone propionate. Findings were that castration reduced both forms of marking, and testosterone therapy to castrated males restored environmental marking in a dose-response fashion. However, the findings for social marking were more complex; for example, a physiological 200-micrograms testosterone dosage to castrated males was unable to elevate conspecific marking over the rates of marking by castrates without testosterone replacement. In experiment 2 the ontogeny conspecific marking in juvenile males was examined in relation to the pubertal surge of androgens. Results suggested that juvenile male marking of an adult male decreased despite a pubertal increase of androgens. Conclusions were that testicular hormones influenced both forms of marking but were less important in the social setting. Moreover, conspecific marking is not simply an extension of marking the environment.     

Disruption of the fifth melanocortin receptor alters the urinary excretion of aggression-modifying pheromones in male house mice.

The preputial glands of house mice express the gene for the fifth melanocortin receptor (MC5-R) and are a primary source of urinary pheromones involved in inter-male aggression. A 'resident-intruder' behavioral model was used to compare the responses of resident males to urine from mice with an engineered disruption of the fifth melanocortin receptor (MC5-RKO) with residents' responses to urine from wild-type mice (WT). Each type of urine was presented in combination with a castrated intruder male to provide the appropriate biological context. Resident males responded with a longer latency to bite when the urine was from gonadally intact WT males compared with urine from MC5-RKO mice. These results are consistent with the hypothesis that activation of the fifth melanocortin receptor in the preputial glands of male house mice causes excretion of urinary pheromones that delay aggressive responses by other males.     

The causation of two scent-marking behaviour patterns in female hamsters (Mesocricetus auratus)

Social, olfactory, and oestrous cycle influences on the frequency of flank-marking and vaginal marking were studied in female hamsters. Vaginal marking was more frequent in the presence of males or their odours than females or their odours. Vaginal marking frequency was greatest the day before oestrus (day 4), intermediate on days 2 and 3, and zero on the oestrous day. The probable sexual advertisement functions of vaginal marking were discussed. In contrast, flank-marking by females was stimulated more by other female odours than by male odours. The frequency of flank-marking was elevated by agonistic encounters and was reduced by sexual encounters. Flank-marking seems to have functions related to functions of aggression among hamsters.     

Assessment of dominance hierarchy through urine scent marking and its chemical constituents in male blackbuck Antelope cervicapra, a critically endangered species

In ungulates the process of chemical communication by urinary scent marking has been directly related to reproductive dominance, territorial defense and proximity to resources. The differences in the frequency of urine marking and chemical composition of urine of males Antelope cervicapra before, during and after the dominance hierarchy period were assessed. The variations in the urine marking and its chemical profiles of dominant males (n = 9), bachelors (n = 5) and sub-adult males (n = 5) were compared to find out how the dominance hierarchy influences the confined blackbuck herd under semi-natural captive conditions. The frequency of urine marking is significantly higher (p < 0.001) in dominant males. Twenty-eight major constituents were identified in the urine of dominant males (before, during and after the dominance hierarchy period), bachelor and sub-adult males. Among these, three specific compounds namely, 3-hexanone (I), 6-methyl-5-hepten-2-one (II) and 4-methyl-3-heptanone (III) were seen only in dominant males urine during the dominance hierarchy period. Based on the behavioural observation and the unique chemical constituents in the urine, it is concluded that the dominant male scent odor suppresses aggression, scent marking, scent production and territorial patrolling activities of subordinate males, through which the dominant male establish their hierarchy and attains success in reproduction.     

The effects of castration and Silastic implants of testosterone on intermale aggression in the mouse

Castration and testosterone (T) replacement were used to study developmental changes in aggressive behavioral responsiveness to androgenic stimulation. Male mice castrated at birth were less sensitive to circulating T than were prepubertal or adult castrates, but fighting was induced in neonatal castrates with a dose of androgen that produced hypertrophy of the accessory organ system in adult castrates. Gonadectomy shortly prior to pubertal increases in serum T concentration also reduced behavioral responsiveness to androgen administration. Intermale aggression was induced in prepubertal castrates only with T treatment that maintained accessory organ growth in adult castrates. The aggressive behavior of males castrated after the pubertal surge in serum T was supported with circulating levels of androgen that failed to stimulate the accessory organ system above that of oil-treated castrates. It was concluded that T stimulation during neonatal or pubertal life is not totally crucial for organization of neural substrates that mediate the ultimate expression of intermale aggression, but exposure to androgen from birth throughout pubertal development is normally required to produce maximal aggressive behavioral responsiveness to circulating T encountered in adulthood.     

The dose-response to testosterone propionate of preputial glands, pheromones and aggression in mice

Urine from female mice injected with 24 daily doses of 0, 1, 10, 50, and 100 μg testosterone propionate (TP) was applied to the coat of castrate males, and the aggressive response of fighter mice toward these castrates recorded. Some antiaggressive property in castrate female urine was confirmed, and this was neutralized by only 1 μg daily TP. Urine from the 50 μg group greatly increased the aggression response, but 100 μg had no additional effect.     

Olfactory Discrimination of Individual Scents in the Subterranean Rodent Ctenomys talarum (tuco-tuco)

Biological odours that convey cues regarding individual identity can provide valuable information mediating many aspects of mammalian social relationships such us dominance hierarchies, group memberships, territorial and mating activities. The ability of the subterranean rodent, tuco‐tuco (Ctenomys talarum), to discriminate between soiled shavings, urine and faeces from different individuals was investigated using the habituation–discrimination paradigm. Discrimination by both males and females was tested using scents obtained from same‐ and opposite‐sex individuals. Each test subject was habituated for three consecutive days to odour samples from the same individual; on the fourth day a scent from a novel individual was provided. For all odour sources, animals spent significantly less time investigating the habituation scent over successive trials, indicating that animals perceived the stimulus as familiar. For all stimuli, test subjects spent more time investigating the novel odour, rather than the familiar one. Animals spent more time investigating soiled shavings than urine or faeces. Both males and females discriminated novel from familiar odours in shavings and urine regardless of the gender of the odour donors. In contrast, test animals discriminated between familiar and novel odours in feces only when the fecal donors were of different sex from subjects. Possible territorial and reproductive functions of individual scent discrimination are discussed.     

Trojan Genes or Transparent Genomes? Sexual Selection and Potential Impacts of Genetically Modified Animals in Natural Ecosystems

Introgression of genetically engineered modifications (GMs) into natural populations represents a new realm for mutation theory. GMs, like mutations, have direct and pleiotropic impacts that can disrupt evolved adaptive suites. If GM males are more competitive or attractive mates, the “Trojan Gene Hypothesis” predicts potentially drastic impacts. We examined sexual selection in transgenic growth hormone (Tg) mice that are strong Trojan candidates given their exceptional size and extensive pleiotropic deficits. We hypothesized that the sophisticated olfactory abilities of females would recognize dysregulation of Tg males (the Transparent Genome Hypothesis). Females expressed interest in Tg males and their volatile scent, but when allowed nasal contact with urine (critical to mate choice) they preferred normal males. Tg male urine had reduced major urinary proteins (important in social signaling) and contained albumin and transferrin indicative of pathology. Novel Tg males failed to elicit pregnancy block in recently inseminated females (the “Bruce Effect”) whereas normal males were highly effective. Normal males expressed high aggression but Tg males were placid, non-aggressive and were largely ignored by normal males. Female mice also strongly preferred normal males over p53^± knockout males in response to volatiles, contact with urine and male presence. This study suggests that conspecific discrimination of fitness may be more powerful than generally appreciated. This has great implications for introductions of GM animals and sexual selection generally.     

The scent of age

In many species, older males are often preferred mates because they carry 'good' genes that account for their viability. How females discern a male's age is a matter of question. However, for animals that rely heavily on chemical communication there is some indication that an animal's age can be determined by its scent. To investigate whether there are changes in body odours with age, and if so their composition, mice were trained in a Y-maze to discriminate urine odours of donor mice of different ages: Adult (3-10 months old) and Aged (more than 17 months old). Trained mice could discriminate between these two age groups by odour alone. To determine the chemical basis for these discriminations, studies were performed using gas chromatography and mass spectrometry. These analyses demonstrated differences in the ratio of urinary volatiles with age. The most prominent differences involved significantly greater amounts of 2-phenylacetamide and significantly lower amounts of methylbutyric acids in Aged animals relative to Adult animals. Fractionating and manipulating the levels of these compounds in the urine demonstrated that the mice can distinguish age based on variation in amounts of these specific compounds in the combined urine.     

The influence of females on male territorial dominance and female preference in dwelling place in laboratory mice

The influence of female mice (Mus musculus) on intermale aggression and the female choice between dominant and subordinate males were tested. Normal male mice were paired with a castrated male or a female. Two pairs of the same kind were housed together after acclimation for 3 days in half (home cage) of the apparatus, which was composed of eight cages connected by tunnels. One of the two normal males became dominant. The dominant males became more aggressive in the presence of females than in the presence of castrated males. The two normal males dwelt in their own home cages. The two females dwelt together and preferred to dwell in the home cages of the dominant male. The two castrated males dwelt together and seemed to decide their dwelling cage more freely than the subordinate normal male. Received: August 20, 1999 / Accepted: December 6, 1999     

Use of olfaction for sexual recognition in the subterranean rodent<Emphasis Type="Italic">Ctenomys talarum</Emphasis>

The ability of the tuco-tucoCtenomys talarum Thomas, 1898 to recognize sex by olfactory cues contained in urine, faeces and soiled shavings was tested by using preference tests. Nonbreeding tuco-tucos selected odours from opposite-sex rather than same-sex conspecifics. This pattern differed between sexes: females spent more time sniffing male than female odours for all scent sources whereas males did not show any difference in the time they spent investigating odours of each sex for each tested odour sources. Dissimilarities in odour selections between sexes may be attributed to a different combination of factors involved in olfactory interest for each sex. The function of gender cues recognition is discussed.     

An observational study of scent-marking in coyotes, Canis latrans

Urination and defaecation patterns of free-ranging coyotes (Canis latrans) were studied in the Grand Teton National Park, Jackson, Wyoming, for two years. The vast majority of urinations by adult males and females were involved in ‘marking’, and differentiating between ‘marking’ and ‘elimination’ may not be necessary. Our results may be summarized as follows: (1) raised-leg urinations (RLU) performed by males were most frequently used in marking. (2) Females marked throughout the year using the squat (SQU) posture. (3) Snow tracking and reading snow sign resulted in a gross underestimate of the relative frequency of SQU's and a large overestimate in the relative frequency of defaecations (DEF) when compared to results obtained by direct observation. (4) There was sexual dimorphism for the contexts in which marking occurred. Overall, marking by males was associated with courtship and mating, with travelling, and with aggression. Marking by females was associated with the acquisition and possession of food and with the denning season. (5) Marking rates per coyote increased in groups larger than two animals. (6) RLU marking rates were greatest in areas of high intrusion when compared to denning areas and areas in which non-group members infrequently tresscent odours are important in orienting individuals in space but do not represent in and of themselves barriers to movement.     

Territorial Behaviour and Social Organization as a Function of the Level of Aggressiveness in Male Mice

Territoriality and social organization were examined in relation to different genetic dispositions for aggressive behaviour. The animals used in the study were male mice of the 51st and 52nd generation of selection for high (Turku Aggressive, TA) and low (Turku Non-Aggressive, TNA) levels of aggressiveness. The level of aggressiveness of the animals was assessed by means of individual tests with non-aggressive standard opponents, after which they were allowed to form individual territories in a 102 times 204 times 90 cm enclosure. TA and TNA males were placed in different enclosures. After 2 wk, when the partitions between individual pens were removed, the behaviour of the animals was observed for a 7-d period. Excessive fighting between the highly aggressive TA males occurred, resulting in the formation of dominant-subordinate relationships. A great number of attacks inflicted were found to be associated with dominance in the colony, and correlated with a high level of aggressiveness when the animals were individually tested for aggression after having been in colony environments. The level of aggressiveness of the TA males that had become subordinates had significantly decreased. The TNA males fought less and were more often found to stay in their original territory for the entire period of observation. The results suggest that different genotypes for aggression arc related to differences in territoriality and social organization in mice.     

Reproductive success in male sunbleak, a recent invasive fish species in the U.K.

The spawning behaviour of male nest guarding sunbleak Leucaspius delineatus , a recent invasive species in southern England, was studied and quantified for the first time. In the absence of physical differences between territorial and non‐territorial males ( i.e . colour, size, etc .), the reproductive behaviour of territorial males was analysed and related to reproductive success. The results showed that females preferred high‐courting and highly aggressive males. The initial cue in female mate choice, however, was based on courtship, while aggression was the decisive behavioural trait in influencing mate choice, providing a direct signal of physical condition and 'paternal competence'. Some males picked nest sites which were subsequently preferred by other males taking over the nest of a previous male ('communal nest'), with the new territorial male adopting the eggs already present at the nest. It appears that either due to female preference for nests already containing eggs or lower rates of sired egg predation by dilution among unrelated eggs, sunbleak males have adopted the mating strategy of allopaternal care.     

Intracranial androgenic and estrogenic stimulation of male-typical behaviors in house mice (Mus domesticus).

Two experiments in house mice (Mus domesticus) examined the neural sites at which steroid hormones activate the following male-typical behaviors: 70 kHz ultrasonic mating vocalizations in response to stimulus females or their urine, urinary marking in response to stimulus males or stimulus females, mounting of estrous females, and intermale aggression. In the first experiment, four groups of castrated males received bilateral intracranial implants of testosterone (T) into either the septum (SEPTUM), medial preoptic area (MPO), anterior hypothalamus (AHA), or ventromedial hypothalamus (VMH). Two control groups received subcutaneous silastic capsules of T (TSIL) or empty silastic capsules (BSIL). The TSIL males performed all behaviors at male-typical levels while the BSIL males were unresponsive. MPO males emitted ultrasonic mating vocalizations at high levels while few vocalizations were seen in males of the other brain implant groups. The VMH, AHA, and MPO males urine marked at higher levels than the BSIL males but did not exhibit the high levels of the TSIL males. Mounting was observed only in MPO and TSIL males. Aggression was rare in males from any of the brain implant groups. In the second experiment, the hormone activity of the implants was increased by using testosterone propionate (TP) or a 50% mixture of estradiol (E2) and cholesterol. The six groups were SEPTUMTP, SEPTUME2, MPOTP, MPOE2, TPSIL, and BSIL. The TPSIL males performed all behaviors at male-typical levels while the BSIL males were unresponsive. TP was effective at restoring vocalizations and urine marking only when placed in the MPO; however, E2 was effective at both sites. Again aggression and mounting were less evident in the brain implanted males. In conclusion, implants of T or TP were effective in restoring ultrasonic mating vocalization when placed into the MPO. MPO implants of T and TP were also effective in stimulating urine marking, although VMH and AHA implants also showed some effectiveness. The restorative effects of E2 were not localized which is probably related to the greater hormonal activity of this treatment. Comparisons of the properties of the various brain implants to restore more than one behavior were discussed.