Male-induced puberty acceleration in young female wild mice: hormonal regulation and source of pheromonal cue

Experiments were designed to examine the influence of adult males on the rate of sexual maturation in young female wild mice. In one experiment, young females were raised in presence of adult males, adult females and in absence of any individual, while in another, they were exposed to urines of: (1) castrated males, (2) spayed females, (3) castrated and TP-treated males, (4) castrated and placebo-injected males. Female maturation as measured by age at vaginal opening and first vaginal oestrus was accelerated by presence of adult males, whereas presence of adult females considerably delayed the vaginal opening and the appearance of first oestrus in young females. In the other set of the experiments, urine from castrated or castrated and placebo-injected males was ineffective in inducing early puberty while urine from spayed females highly delayed the sexual maturation. By contrast, urine from castrated and TP-treated males accelerated the puberty more or less like normal males. The results indicate that male's chemosignal accelerating puberty in young females is present in urine and its production is under the control of androgens. However, the female-originating urinary pheromone which delays the puberty in young females is not regulated by ovarian hormones.     

Effects of urine from females in oestrus on puberty in female mice

A series of 9 experiments was conducted to examine various characteristics of the urinary chemosignal found in the urine of oestrous female mice that accelerates the sexual development of conspecific females. This urinary chemosignal was effective in doses as small as 0.001 ml/day, was present in excreted and bladder urine, required 3 days of treatment starting before Day 29 of age to effect an acceleration of puberty, required a minimum daily exposure of 2 h, and was relatively nonvolatile. In addition the chemosignal from oestrous females was effective in summer but not in winter months, was significantly more effective when collected at the middle or end of the dark portion of the daily cycle than at the beginning of the dark phase or middle of the light phase, and was not affected by food deprivation or shortened photoperiod. Simultaneous treatment of test subjects with urine from oestrous females and grouped females resulted in delays in puberty and simultaneous treatment with urine from oestrous females and urine from males or pregnant or lactating females did not result in any enhanced acceleration of puberty.     

A study of spontaneous sexual maturation of the female ferret

Although ferrets are long-day breeders, females reared exclusively in nonstimulatory short days will undergo spontaneous sexual maturation by 30-50 wk of age. In the following report, this spontaneous sexual maturation of ferrets was studied to determine mechanisms regulating sexual maturation in nonstimulatory photoperiods. Study of ovariectomized females treated with low, constant levels of estradiol suggest that a marked decrease in the efficacy of estradiol to inhibit luteinizing hormone secretion occurs shortly before sexual maturity becomes evident in intact controls (both groups housed in short days). During this long juvenile period, a marked increase in body weight occurred, but ovarian responsiveness to exogenous gonadotropin did not change. Older, larger females did respond more rapidly to stimulatory photoperiod than did younger females. These studies suggest that the mechanisms of spontaneous puberty in ferrets are likely the same as those regulating photoperiod-stimulated puberty in this species.     

Effects of donor age on urinary chemosignals that influence the timing of first oestrus in young female house mice, Mus domesticus

The effects of donor age on the effectiveness of puberty-influencing urinary chemosignals in wild house mice was tested in a series of 3 experiments. The chemosignal from male mice that accelerates puberty was present in the urine from about the time of puberty and throughout the normal lifespan, but declined about 1 year of age. Oestrous females released a substance in their urine that accelerates puberty in young females. This substance remained effective from first oestrus until over 1 year of age, although older females were in oestrus less frequently than younger mice. Females that are pregnant or lactating released a puberty-accelerating substance in their urine regardless of age. Production and release of the puberty-delaying chemosignal by grouped females was initiated before puberty and continued throughout the lifespan of the mouse.     

Hormonal and behavioral changes at puberty in the squirrel monkey

Developmental changes in the reproductive behavior and physiology of 9 male and 15 female juvenile squirrel monkeys were evaluated in a 20-month study. Plasma levels of gonadal steroids remained relatively low for this species until most animals reached puberty between 2.5 and 3 years of age. Longitudinal assessment of plasma progesterone levels indicated that the onset of ovarian cycles tended to be synchronized between females although the 5 heaviest females began to cycle earlier than the rest. The heavier females reached puberty at a time which was appropriate to their birth in the wild, whereas most of the remaining females conceived 6 months later during a second period of reproductive activity that coincided with the laboratory mating season. Pubescent males underwent their first seasonal elevation in plasma testosterone levels during the second period and its onset was synchronized across all males. Thus, even in the absence of adults, pubertal processes in the squirrel monkey were strongly influenced by the seasonal breeding pattern. In addition, behavioral observations revealed that social maturation closely parallels reproductive ability in females, whereas males enter a protracted subadult stage after puberty.     

Acceleration of puberty in female mice by a chemosignal from pregnant and lactating females: circadian rhythm effects

Two experiments were designed to test whether the urinary chemosignal excreted by pregnant and lactating female mice that accelerates puberty in young females is affected by circadian rhythms. The experiments also measured the possible influence of circadian rhythms on the response of the young recipient females. For urine from both pregnant and lactating females there was no difference in the effectiveness for accelerating puberty in urine collected during all 24 h. However, pregnancy urine used for treatment at 1800 and 0000 h, and lactation urine used for treatment at 1800, 0000 and 0600 h, all resulted in significantly earlier mean ages for puberty than pregnancy urine treatment at 0600 or 1200 h, or lactation urine treatment at 1200 h. There was also a significant interaction between the time of urine collection and the time of urine treatment for each urine source; urine was generally more effective in accelerating sexual development when used for treating young females at the same hour at which it had been collected, or at the time interval(s) just before or after the time at which it had been collected.     

Acceleration and delay of first vaginal oestrus in female mice by urinary chemosignals: Dose levels and mixing urine treatment sources

The effects of varying dose levels and mixing of urine from various types of donor mice on the age of sexual maturation in female mice were tested. Over the range from 0.001 ml/day to 0.01 ml/day, there was no difference in the effectiveness of male urine in producing acceleration of puberty, nor was there any difference over the same dose range for urine from grouped females bringing about a delay of puberty. Urine from pregnant and lactating females brought about earlier puberty when applied in the higher dose amounts but was not effective in altering the age of first oestrus relative to untreated controls at lower doses. These findings concerning dose levels are important for a full understanding of the behavioural consequences of urinary chemosignals. When urine from different sources was mixed, all treatments which involved urine from grouped females produced delays in first oestrus. The second finding has important consequences for a feedback model for population regulation in house mice involving urinary chemosignals that accelerate or delay sexual maturation and thus shorten or lengthen generation time by affecting reproductive behaviour.     

Effect of density, duration of grouping and age of urine stimulus on the puberty delay pheromone in female mice

The ability of urine from female mice to delay puberty in test females was directly related to the density and duration of grouping of females. When females were removed from group housing their urine lost its ability to delay puberty within 10 days. No interactive effects were observed between duration and density of grouping on the onset of pheromone release after grouping or on the persistence of pheromone release after re-isolation. Urine from grouped females lost its ability to delay puberty in test females after 7 days of exposure to air.     

Aggressive behavior in female golden hamsters: development and the effect of repeated social stress

In male golden hamsters, agonistic behavior matures during puberty, changing from play fighting to adult-like aggression. In addition, this transition is accelerated by repeated social subjugation early in puberty. However, little is known about the development of agonistic behavior in females. In the present study, we compared the development of agonistic behavior in male and female golden hamsters. Furthermore, we also tested the effects of repeated social subjugation on the development of agonistic behavior during puberty. Hamsters were tested for agonistic behavior in the presence of a smaller intruder at different intervals during puberty. Several observations were made. First, the frequency of attacks remained stable in females, while varying in males. Second, the transition from play fighting to adult-like aggression occurred at earlier time periods in females than in males. Finally, a clear transitional period marked by attacks focused on the flanks was observable in males around mid-puberty. However, this transitional period was not apparent in females. In addition, juvenile females were exposed to aggressive adult males or females. In both cases, repeated exposure to stress had no statistically significant effect on the development of agonistic behavior. After 2 weeks of subjugation, exposure to aggressive adults had no effect on serum cortisol levels, indicating that juvenile females habituate to repeated social stress. These data show significant sex differences in the development of agonistic behavior and adaptation to repeated stress in juvenile golden hamsters.     

Urinary sex steroids during sexual development in female mice and in proximate novel males.

The experiments described here were designed to determine whether males' capacity to accelerate female pubertal development is reflected in females' urinary steroid levels in mice, and whether steroids in males' urine are influenced by exposure to developing females. In the first experiment, measures from urine collected daily from female mice aged 31-59 days showed a gradual rise in 17beta-estradiol levels and a distinct linear rise in progesterone levels. In a second experiment, daily steroids were measured in females aged 30-42 days while they were either housed alone or underneath two novel outbred males. Females exposed to males showed accelerated development at day 43 in uterine weight, and to a lesser extent in ovarian and whole-body weights. Average steroid levels did not significantly differ between conditions, but intra-individual variance in estradiol measures was greater in male-exposed than in isolated females. Creatinine levels were higher in isolated females. Males exposed to developing females excreted higher levels of estradiol in their urine compared to isolated males. These data suggest that excreted steroids can reflect general pubertal development, but may not fully reflect substantial morphological impacts of exposure to novel males. Elevations of estrogen levels in males exposed to developing females could help to account for precocious puberty in such females.     

Body weight as an effective tool for determination of onset of puberty in captive female Nile hippopotami (Hippopotamus amphibious)

Profiles of fecal progestogens and body weight from the early juvenile to the peri‐pubertal period are presented for eight captive female Nile hippopotami housed at Disney's Animal Kingdom in Florida. Average growth rate in juveniles was 0.85±0.03 kg/day (r²=0.913). Progestogen elevations were detectable as early as 16.8 months of age, and elevations indicative of ovulation and luteal activity were identified in seven of eight females by 30.3±1.6 months of age and body weight of 829.3±49.4 kg. Progestogen patterns before the onset of puberty were highly variable within and between females. Some females remained at baseline concentrations, whereas in others the progestogen pattern was characterized by infrequent, transient elevations of low amplitude and shortened duration. Four females were monitored through onset of puberty, conception, and first pregnancy. Onset of puberty was defined as the first luteal phase from the series of consecutive ovarian cycles culminating in conception and was observed at 34.9±2.2 months of age and 963.6±39.4 kg, however, the quality and number of cycles varied among females before conception. Females conceived between 2.7–3.9 years of age after attaining an approximate threshold body weight of 1,000 kg (1,070.5±39.5 kg). The age at first conception in captivity occurs at a younger age than has been reported for wild populations. Body weight may be an effective tool for approximating the state of reproductive maturity and facilitate collection management in zoos. Zoo Biol. 0:1–13, 2005. © 2005 Wiley‐Liss, Inc.     

Puberty delay of bank vole females in a high-density population

The onset of puberty in bank vole females was studied, with uterine weight, ovarian weight, and the number of large ovarian follicles used as indicators of gonadal activity. Maturation of females born at the beginning of the reproductive season was suppressed by the presence of other females. Puberty of animals born at the end of season was primarily influenced by climatic variation.     

Puberty in pine voles, Microtus pinetorum, and the influence of chemosignals on female reproduction

We investigated the reproductive biology of an induced ovulator, the pine vole (Microtus pinetorum). Male puberty, measured as age at first impregnation, was found to occur as early as 44 days of age. Female puberty measured as age at first conception, was found to occur as early as 32 days of age, considerably earlier than previously reported. Females paired with stud males exhibited a doubling of uterine weight within 12 h, and vaginal sperm were present after 48 h. This indicates that although behavioral responses to males--including mating--require prolonged contact, physiological responses to males occur rapidly. Chemosignals from males slightly increased uterine and ovarian weights of females, but chemosignals from other females did not. Young females paired with stud males for 48 h in the presence of soiled bedding from the female's family had significantly smaller increases in ovarian and uterine weights than similar females paired on clean bedding. Suppression of reproduction in female offspring while they remain with the extended family unit is discussed as a life-history tactic and as a possible mechanism for inbreeding avoidance.     

Lack of pubertal influences on female dispersal in muriqui monkeys, Brachyteles arachnoides

The hormonal mediation of dispersal in female mammals is poorly understood, in part because of the difficulties of detecting the onset of ovarian cycling and puberty in dispersing individuals. We used noninvasive methods of faecal steroid assays to determine the timing of dispersal relative to puberty and ovarian cycling in wild female muriqui monkeys, a species in which males are philopatric and nearly all females transfer from their natal groups. Natal females had a mean+/-SE age of 73.4+/-7.2 months (N=18) at the time of their transfers. Intergroup transfers occurred when one or more sexually active adult females were present, but did not show any seasonal patterns. Faecal progesterone and oestradiol profiles from nine natal females prior to transfer and four non-natal females that transferred into our study group demonstrate unequivocally that dispersal occurs prior to puberty in this species. All females showed baseline oestradiol levels and low progesterone levels compared with cycling adult females. Immigrants were first observed to copulate at 11.2+/-2.2 months of age (N=4), prior to the onset of normal ovarian cycles, and gave birth to their first offspring at 33.8+/-7.3 months (N=4) after transferring. Mean cortisol levels did not differ between natal emigrants or recent immigrants, and were within the range of those of adult males during the nonbreeding season in 10 of the 11 prepubertal females sampled. These results indicate that female dispersal is not triggered by activational hormones associated with puberty or escape from reproductive suppression in this species. Copyright 2000 The Association for the Study of Animal Behaviour.     

Long-term effects of accelerated or delayed sexual maturation on reproductive output in wild female house mice (Mus musculus)

The effects of acceleration and delay of puberty in female house mice on survival and reproduction were tested using 6 experimental groups: (1) control females mated at the time of first oestrus, (2) females mated at weaning, (3) females treated with male urine starting at weaning and mated at first oestrus, (4) females housed in groups and mated at first oestrus, (5) females housed alone, treated with urine from grouped females and mated at first oestrus, and (6) females housed alone and mated at 68 days of age. Females caged with males at weaning or treated with male urine and mated at puberty had lower rates of survival to 180 days of age, but did not differ in rates of fertility from mice in the other four treatments. Those females that were housed with males from weaning or treated with male urine also had smaller total numbers of litters, fewer total young, and smaller average litter sizes than did females for which the age of mating was delayed, by grouping or treatment with urine from grouped females, or by being held until age 68 days before mating. Control females mated at first oestrus generally were intermediate or did not differ from the male treatments on these dependent variables. There were no differences in the average number of female young/litter across the 6 treatments. However, females that were delayed in age of first mating had significantly more male young/litter than did females that were accelerated in their sexual development or control females.(ABSTRACT TRUNCATED AT 250 WORDS)     

MPOA cytotoxic lesions and maternal behavior in the rat: effects of midpubertal lesions on maternal behavior and the role of ovarian hormones in maturation of MPOA control of maternal behavior

Small neurotoxin lesions in the medial preoptic area (MPOA) block maternal behavior (MB) in adults but large lesions are required to produce the same effect in juvenile rats (23-27 days of age). To study the maturation of MPOA control of MB, in Experiment I, we compared the effects of small versus large neurotoxin MPOA lesions at midpuberty (38 days of age) on MB. Midpubertal females with large MPOA lesions showed severe impairment in MB affecting retrieving, crouching, and nest building, but 85% of females with small MPOA lesions exhibited all components of MB and performed like control females without MPOA lesions. To study the role of ovarian hormones during puberty on the maturation of MPOA mediation of MB (Experiment IIA), females were ovariectomized either before or after puberty and small MPOA cytotoxic lesions were made at 53 days of age. At 60 days of age both groups showed similar deficits in MB which indicated that the maturation of the MPOA mediation of MB is not dependent on pubertal ovarian hormones. In Experiment IIB, we administered estradiol benzoate (sc) and this overcame the deficit in MB after small MPOA lesions in females that had been deprived of estrogen for shorter periods (30 days) but had not been deprived for longer periods (60 days). In addition, ovary-intact females with circulating estrogen and small lesions in the MPOA at 53 days of age did not show deficits in MB.     

Intermittent stimulation and acceleration of puberty by urinary chemosignals in female mice (Mus musculus domesticus)

A sequence of 17 experiments was used to test the effects of intermittent stimulation with urinary chemosignals on the age of puberty in young female mice. The three chemosignals tested all accelerate the age of sexual maturation: urine from adult males, urine from females in estrus, and urine from females that are pregnant or lactating. The basic technique involved presenting the prepubertal females with 'Nestlets' on which the urine was placed. The 'Nestlets' were placed in the cages of the test females for a 15-min period, removed for a variable period, and then replaced in the cage for 15 min. In this manner it was possible to vary the number of exposures, the total length of exposure, and the total time period over which the exposures occurred. Control procedures, involving exposures of young females to cotton squares with water rather than urine placed upon them, resulted in no alterations in puberty relative to untreated females. For mice exposed to the urine-treated cotton squares, acceleration of puberty occurred with less total stimulus-exposure time when the stimulus was presented in short exposures over a number of hours than in previous investigations when the exposure to the urinary chemosignal occurred in a single block of time of one or two hours. For each of the three acceleratory chemosignals, there was a diminution of acceleratory effect when the ratio of total stimulus-exposure time to total exposure time grew smaller. This diminution was more pronounced for urine from pregnant or lactating females than for urine from males or from females in estrus.(ABSTRACT TRUNCATED AT 250 WORDS)     

Salivary and urinary metabolome analysis for pre-puberty-related biomarkers identification in porcine

Estrus synchronization is important for optimal management of gilt reproduction in pig farms. Hormonal treatments, such as synthetic progestogens, are used on a routine basis, but there is a growing demand for non-hormonal alternative breeding tools. Before puberty, gilts exhibit a ‘waiting period,’ related to the ovarian development and gonadotrophin secretions, during which external stimulations, such as boar exposure, could induce and synchronize first ovulation. Practical non-invasive tools for identification of this period in farms are lacking. During this period, urinary oestrone levels are high, but urine sampling is difficult in group-housed females. The aim of this work was to search for specific biomarkers of the ‘waiting period’ in saliva and urine. In total, nine 144- to 147-day-old Large White gilts were subjected to trans-abdominal ultrasonography three times a week for 5 weeks until puberty detection (week –5 to week –1 before puberty). Urine and saliva samples were collected for oestrone assay to detect the ‘waiting period’ and for metabolome analysis using ¹H-nuclear magnetic resonance spectroscopy to detect potential biomarkers of the ‘waiting period.’ Gilts were slaughtered 7 days after puberty detection for puberty confirmation. Results were consistent with ultrasonography data for six gilts. Urine and saliva samples from these six gilts were analyzed. Urinary estrone concentration significantly increased 2 weeks before puberty detection. Metabolome analysis of urine samples allowed the identification of 78 spectral bins, among them, 42 low-molecular-weight metabolites were identified. Metabolome analysis of salivary samples allowed the identification of 59 spectral bins, among them, 23 low-molecular-weight metabolites were detected and 17 were identified. No potential biomarker was identified in urinary samples. In saliva, butyrate and 2HOvalerate, 5.79 ppm (putatively uridine), formate, malonate and propionate could be biomarker candidates to ascertain the pre-puberty period in gilt reproduction. These results confirm that non-invasive salivary samples could allow the identification of the physiological status of the gilts and presumably the optimal time for application of the boar effect. This could contribute to synchronize puberty onset and hence to develop non-hormonal breeding tools.     

Graded response to short photoperiod during development and early adulthood in Siberian hamsters and the effects on reproduction as females age

Short day (SD) lengths delay puberty, suppress ovulation, inhibit sexual behavior, and decelerate reproductive aging in female Siberian hamsters (Phodopus sungorus). To date, the modulation of the age-associated decline in reproductive outcomes has only been demonstrated in female hamsters experiencing different day lengths during development. To determine if developmental delay is necessary for photo-inhibition to decelerate reproductive aging, hamsters raised in LD were transferred to SD as young adults and remained there for 6 months. Females that demonstrated the most immediate and sustained photo-inhibition were found to have greater numbers of ovarian primordial follicles at advanced ages (9 and 12 months) than did females held in LD, nonresponders to SD, and females with a marginal SD-response. Similarly, for females raised in SD from conception to 6 months of age, prolonged developmental delay was associated with greater numbers of primordial follicles at later ages as compared to hamsters that became refractory to SD. A robust response to SD in juvenile and adult hamsters is associated with decelerated reproductive aging, which may result in greater reproductive success in older females as compared to age-matched individuals demonstrating a more modest response to SD.     

Analysis of male pheromones that accelerate female reproductive organ development

Male odors can influence a female's reproductive physiology. In the mouse, the odor of male urine results in an early onset of female puberty. Several volatile and protein pheromones have previously been reported to each account for this bioactivity. Here we bioassay inbred BALB/cJ females to study pheromone-accelerated uterine growth, a developmental hallmark of puberty. We evaluate the response of wild-type and mutant mice lacking a specialized sensory transduction channel, TrpC2, and find TrpC2 function to be necessary for pheromone-mediated uterine growth. We analyze the relative effectiveness of pheromones previously identified to accelerate puberty through direct bioassay and find none to significantly accelerate uterine growth in BALB/cJ females. Complementary to this analysis, we have devised a strategy of partial purification of the uterine growth bioactivity from male urine and applied it to purify bioactivity from three different laboratory strains. The biochemical characteristics of the active fraction of all three strains are inconsistent with that of previously known pheromones. When directly analyzed, we are unable to detect previously known pheromones in urine fractions that generate uterine growth. Our analysis indicates that pheromones emitted by males to advance female puberty remain to be identified.