Effects of prior androgen exposure upon adrenal function in maturing rats

Gonadectomy in the newborn produced an elevated or “female” pattern of serum corticosterone levels when basal levels of corticosterone (B) were measured at the trough and peak of the daily rhythm and when the response to a 2 minute ether stress was recorded after weaning age. Administration of 1.25 mg testosterone propionate (TP) at two days of age did not significantly alter corticosterone patterns in intact males but did prevent the rise in serum B noted in gonadectomized (Gondx) males. In contrast with oil-injected females, females androgenized with 1.25 mg TP showed precocious vaginal opening,persistent estrous and a failure of serum B to become elevated at noon on the day of vaginal opening. Gonadectomy at weaning age had no effect on basal levels of B but there was a more rapid elevation of B after ether stress in Gondx males. By 30 minutes post-stress, the levels of B were the same in both groups. Gonadectomy at 60 days of age had no effect on B levels except for a transient decrease in B at 8 p.m. one week after surgery. The stress response was also normal in castrated males. It can be concluded that early androgen exposures does alter the functional properties of the adrenal but that continued secretion of androgen is not necessary to maintain adrenal function at a “male” level in adults.     

Adrenal function in female rats before puberty: the effects of gonadectomy

The following study was designed to test whether the change in the amplitude of the adrenal rhythm and the response to stress seen at puberty in female rats is dependent upon stimulation of the adrenal system by gonadal steroids. Rats were gonadectomized either at two days of age or at 21 days of age and periodic blood samples were taken by cardiac puncture and assayed for corticosterone (B) by a fluorometric procedure. Ovariectomy at weaning age (21 days) had no effect on adrenal function until the time of normal puberty onset. In intact animals, a rise in resting levels of corticosterone and an increase in the incremental response to ether stress was noted at 35 days of age. (Puberty in females was 35.3 ± 1.2 days as indicated by vaginal opening). Ovulation occurred the following day in 9/10 rats. Gonadectomy at two days of age had essentially the same effect as later gonadectomy in females. By 70 days of age, resting corticosterone values and stress responses in gonadectomized females reached levels similar to intact females. It is concluded that an independent adrenarche can occur in females but that gonadal steroids present at the time of puberty modulate the timing of this process.     

Age- and hormone-related changes in vaginal smear patterns in the gray-tailed vole, Microtus canicaudus

Female voles, Microtus canicaudus, exhibited age-related changes in vaginal smear patterns when isolated from males after weaning. Between 30 and 50 days of age, nearly all females exhibited persistently leucocytic vaginal smears. By 90-120 days, most females showed vaginal cyclicity with alternating predominance of leucocytes, nucleated epithelial cells or cornified epithelial cells. Most females examined between 150 and 200 days of age exhibited persistent vaginal cornification. The vaginal cyclicity seen in females between 90 and 120 days was not a reflection of cyclic ovulatory changes; plasma progesterone concentrations remained constant, regardless of age or vaginal smear pattern, and corpora lutea were never seen in unmated females. Although progesterone concentrations did not differ among vaginal smear patterns of 120-day-old females, plasma oestrogen values were highest in females exhibiting vaginal cornification.     

Age at maturity in cavies and guinea-pigs (Cavia aperea and Cavia aperea f. porcellus): influence of social factors

Age at maturity, a particularly important parameter in the life history of small mammals, contributes greatly to fitness. Social influences on age at maturity have been demonstrated for altricial rodents, in particular, mice. Nothing is known about such effects in precocial small mammals. Wild cavies Cavia aperea are born in a highly precocial state and mature early in life, briefly after weaning. We investigated whether the wild cavy C. aperea and the domestic guinea-pig Cavia aperea f. porcellus reach maturity earlier in the presence of adults of the opposite sex. Juvenile females kept in pairs without males showed first vaginal opening (=oestrus) when 59 days old in cavies and at about 40 days in the guinea-pig. However, in the company of adult males, cavy females kept in pairs reached maturity when about 30 days old, and guinea-pig females when 26 days old. Most cavy females experienced successful pregnancy following first vaginal opening. In cavies, female mass at birth and at first oestrus was not correlated with age at first oestrus. In guinea-pigs, birth mass predicted age at maturity only when a male was present. The growth rate from birth to first oestrus related to age at first oestrus. In the wild cavy, the presence of a male appeared to influence maturation more between days 25 and 30 than earlier in life. Male C. aperea matured and had fully descended testes when about 65–70 days old. All male cavies produced abundant motile sperm from day 75. First successful copulations occurred at about the same age. Surprisingly, the priming effect of the presence of an adult male on female maturation proved stronger in these highly precocial caviomorphs than in altricial rodents investigated so far.     

Transitory increase in plasma FSH and LH levels induces early vaginal opening and corpus luteum formation in ovarian transplants of rats ovariectomized at 20 days of age

The effect of a transitory increase in plasma FSH and LH levels on puberty has been investigated. Hormonal changes were induced by bilateral ovariectomy (OVX) at 20 days of age followed by implantation of two ovaries (donor animals 20-day-old) beneath the kidney capsule 1, 2 or 3 days later. In an other group of animals the two interventions were made in reversed order. Rats in which OVX was made first showed early vaginal opening. If ovarian implantation preceded OVX, these two interventions did not affect the time of vaginal opening. More than 1/3 of the ovarian implanted animals exhibiting early opening of the vagina contained corpora lutea three days after vaginal opening. FSH and LH levels of OVX + ovary implanted rats were from the third day on following implantations markedly different from the OVX controls and were in the range of intact or sham-operated rats. The present findings indicate that a transitory increase in plasma gonadotropin levels around twenty days of age may lead to precocious puberty.     

Pulsatile infusion of luteinizing hormone-releasing hormone induces precocious puberty (vaginal opening and first ovulation) in the immature female guinea pig

Previously we have hypothesized that an increase in luteinizing hormone-releasing hormone (LHRH) due to hypothalamic maturation is the key factor controlling the onset of puberty. This led to the working hypothesis that precocious puberty would be induced if LHRH is administered with an appropriate protocol. Thus, effects of pulsatile infusion of LHRH on the onset of first vaginal opening and first ovulation in immature female guinea pigs were studied. Luteinizing hormone-releasing hormone in hourly pulses of either 5 ng or 50 ng was infused through a chronically implanted jugular catheter for 9-29 days starting at 20 days of age. For the control experiment saline was infused in a similar manner. Infusion of 5 ng LHRH/h resulted in significantly earlier (P less than 0.001) ages at first vaginal opening (24.7 +/- 0.9 days) and at first ovulation (28.8 +/- 0.9 days) compared to saline controls (first vaginal opening 53.3 +/- 6.8 days; first ovulation 55.2 +/- 6.5 days). Infusion with a 10-fold higher LHRH dose (50 ng/h) also advanced the age at first vaginal opening (25.3 +/- 0.7 days), but precocious ovulation was no longer induced (53.7 +/- 5.3 days). Interestingly, LHRH infusion with the high dose resulted in a prolonged period of vaginal opening and cornification without ovulation. These results indicate that 1) pulsatile infusion of a small amount of LHRH with a constant frequency induces precocious puberty in a laboratory rodent, and 2) infusion of LHRH with a dose higher than the effective dose for the induction of early puberty results in a persistent estrous anovulatory syndrome. Therefore, the present study not only supports our hypothesis that an increase in endogenous LHRH release is responsible for the onset of puberty, but also further suggests that excessive release of LHRH or abnormal patterns of LHRH release may be involved in the etiology of the anovulatory persistent estrus syndrome.     

Urogastrone-epidermal growth factor and aspects of sexual maturation in female rats as a function of age at treatment.

The effect of epidermal growth factor (EGF) on sexual maturation of female rats was studied. A within-litter experimental design was employed, so that in each litter each female received four daily injections of EGF (E, 500 ng/g body weight s.c.) or vehicle (V), at one of three ages: days 0-3 (E1, V1), days 8-11 (E2, V2), days 16-19 (E3, V3). Body weight, pinna detachment, incisor eruption, eye opening, auditory startle, visual placing, vaginal opening and first cytological oestrus were assessed. Neonatal treatment with EGF (E1) delayed pinna detachment and the appearance of the auditory startle, but accelerated eye opening. Also, E1, but not E2 and E3, resulted in lower body weight at weaning. Treatment E3 advanced sexual maturation, as indicated by vaginal opening and first cytological oestrus, by 5-6 days. E1 and E2 had no such effect. Hence the sensitive period for the effect of EGF on female rat sexual maturation is later than that for effects on other developmental characteristics and body weight. In a second experiment, ovary and uterus weights were found not to differ between E3 and V3 females killed on the day of vaginal opening of the E3 rats, suggesting that the effect of EGF may be specifically on the perineal epithelium and not on sexual maturation generally.     

Hormonal Control of Sexual Differentiation of the Hypothalamus in the Neonatal Female Rat

Groups of female rats were treated either on the day of birth or at 5 days of age with oil, testosterone propionate (TP), MER-25 (an oestrogen antagonist) or TP plus MER-25. Treatment with oil and MER-25 alone on day 1 or 5 did not prevent the development of cyclic repro ductive activity in any of the females. Administration of TP on the day of birth inhibited vaginal opening and prevented cyclic ovarian activity. Treatment with TP on day 5 significantly advanced the time of vaginal opening but prevented both vaginal and ovarian cyclicity. Treatment on the day of birth with a combination of MER-25 and TP did not significantly prevent the masculinizing action of TP. In most cases vaginal opening failed to occur; the ovaries were small and devoid of corpora lutea. Sexual receptivity was not significantly affected by treatment on the day of birth with oil, TP or MER-25 alone. When MER-25 was given before the TP, however, receptivity was significantly enhanced compared with oil treatment. At 5 days of age treatment with MER-25 plus TP prevented the masculinizing action of TP. Most of the animals had normal ovarian weights, and ovaries which contained both follicles and corpora lutea. The most effective treatment for preventing the action of TP was administration of MER-25 6 h before the injection of androgen. After treatment at 5 days of age, sexual receptivity was significantly reduced in all groups compared with the oil-treated animals. Administration of MER-25 plus TP did not enhance sexual receptivity above that found in the animals receiving TP alone. The results are discussed in terms of the possible role of oestrogen in controlling hypothalamic differentiation in the neonatal female rat.     

In utero contiguity to males does not influence morphology, behavioral sensitivity to testosterone, or hypothalamic androgen binding in CF-1 female mice.

Physiological and behavioral systems presumably influenced by prenatal exposure to testosterone (T) were compared in CF-1 female mice from known uterine positions. Anogenital distance did not differ among females that developed in utero between two females (0M), adjacent to one male (1M), or between two males (2M) at birth, at weaning on Day 21, or on Day 60 postpartum. The age of vaginal opening and mean estrous cycle length also were similar among the groups. When ovariectomized and implanted with a T-containing silastic capsule, the mean number of days of treatment required to activate male-like aggressive behavior also did not differ among the three positional classifications. Finally, androgen binding in combined hypothalamic-preoptic-septal cytosol was assessed after 8 days of T treatment, and no systematic variation in [3H]DHT binding related to uterine position was found. These results indicate that contiguity to male fetuses did not induce variation among CF-1 females in morphological, behavioral, or biochemical systems thought to be influenced by prenatal exposure to T.     

Relationship of vaginal impedance with speed of return to oestrus after weaning, oestrous behaviour, parity and lactation length in cyclic sows

The effect of weaning to oestrus interval, oestrus duration, parity, lactation length, breed and their interactions on changes of vaginal impedance in sows after weaning and during oestrus was examined. The impedance measurements were carried out by a four-electrode method. The interval from weaning to oestrus was significantly longer in sows with the length of lactation 21-25 days than 26-30 days and 31-36 days and in primiparous than multiparous sows. The interval from weaning to oestrus was negatively correlated with the length of lactation (r=-0.21; P<0.05), parity (r=-0.36; P<0.01) and oestrus duration (r=-0.26; P<0.01). The weaning to oestrus interval, oestrus duration, parity and lactation length had a significant effect and the breed of sows had no influence on the vaginal impedance in peri-oestrus. The decrease of vaginal impedance after weaning was delayed in sows with a longer weaning to oestrus interval and in primiparous than multiparous sows. The decline of vaginal impedance during peri-oestrus was more gradual in sows with a longer interval from weaning to oestrus, shorter lactation, primiparous sows and sows with the length of oestrus 36 h and 72 h and more. The nadir of vaginal impedance occurred earlier before oestrus in sows with a shorter oestrus. The interaction of weaning to oestrus interval with parity and oestrus duration and the interaction of oestrus duration with parity significantly affected the vaginal impedance in weaned sows. In conclusion, the weaning to oestrus interval, oestrus duration, parity and lactation length considerably influence the vaginal impedance in sows during peri-oestrus. The findings indicate that the impedance technique may be a useful method for a study of factors and processes that accelerate or slow down the return to oestrus after weaning and affect oestrus duration in sows.     

Relation between body weight, growth rate, chronological age and puberty in male and female rats

Body weight, growth rate, chronological age and puberty in female and male rats from litters of 8 and 12 offsprings/mother have been studied. Age and body weight at the moment of vaginal opening (VO) and balanopreputial separation (BPS) were analyzed. Results show that animals reared in smaller groups grew faster than others. After weaning there was an increase in growth rate. VO and BPS occurred at the same age in groups with different growth rates and different body weights. In conclusion this work evidences that external signs of sexual maturation are not linked to a "critical weight" or to a "growth rate".     

A novel postcopulatory block of reproduction in white-footed mice

Female white-footed mice (Peromyscus leucopus) fail to produce offspring when paired with a male from weaning until 150 days of age if an adult female or her odor is also present. The present study delineates more clearly which stage of the young female's reproductive cycle is inhibited by the chemosignal of the older female. Age at vaginal opening and first estrus are delayed by the presence of the older female, but only for about 10 days. The presence of a male counteracts this delay. Thus, the basis for the block is not a delay in puberty. The older female's presence does not influence the number of estrous cycles experienced during the 30 days following first estrus, nor does it influence the number of corpora lutea found at autopsy. The presence of sperm in the vaginae of young females indicates that they were copulating. Likewise, examination of embryos 2 and 3 days after copulation reveals normal developmental progress. However, implantation does not occur in young females that have been exposed to an adult female. Thus, the block occurs either during the final stages of embryo transport or in relation to the implantation process itself.     

The influence of prenatal and postnatal fraternity size on reproduction in mice

Effects of prenatal and postnatal fraternity size (size of litter in which an individual is reared) on age at vaginal opening, growth, and subsequent litter production were examined by rearing mice in standardized prenatal and postnatal fraternities in a 3 X 3 factorial design. Prenatal fraternity size was standardized by reducing litters of 14 or more to either 14, 10, or 6 fetuses on Day 9 of gestation. Postnatal fraternity size was standardized by assigning pups randomly to nurse in litters of 5, 10, or 15 pups within 24 h of birth. Both prenatal and postnatal fraternity size affected growth of the mice (p = 0.02 and p less than 0.01, respectively) with mice reared in small fraternities attaining greater weights throughout the study. Prenatal fraternity size had a negative linear effect on litter size at second parity and tended to have a positive linear effect on age at vaginal opening. Postnatal fraternity size affected age at vaginal opening (p less than 0.01), litter size at first parity (p less than 0.01), and litter size at second parity (p = 0.03). These effects were positive and linear for age at vaginal opening (p less than 0.01; small fraternity size associated with younger age at vaginal opening) and negative and linear for litter size at first and second parity (p less than 0.01 and p = 0.10, respectively; smaller fraternity size associated with larger litter size). There was no interaction between prenatal and postnatal fraternity size effects on age at vaginal opening or litter size (p greater than 0.20).     

Perinatal ACTH-naloxone treatment: Effects on physical and behavioral development

Pregnant mice were treated with either ACTH or naloxone, or both, from 2 to 5 days prior to parturition, until 5 days postpartum. Perinatal ACTH caused a retardation of growth, eye opening, and vaginal opening that was blocked by naloxone; perinatal naloxone produced a decrease in open field activity at 32 days of age that was blocked by ACTH; naloxone reduced the threshold for heat at 50 days of age with no interaction with the ACTH treatment. It is proposed that part of the role of ACTH in physical systems may be mediated via naloxone-sensitive receptors. The direct effect of naloxone on another class of receptors may mediate the changes in threshold to heat flinch. Effects of ACTH and naloxone on the mother may have indirectly produced alterations in physical and motor development of the offspring.     

Activation of AMPA receptors inhibits prolactin and estradiol secretion and delays the onset of puberty in female rats

Previous experiments have evidenced the neuroendocrine role of AMPA receptors. Present studies were carried out to obtain information on the role of these receptors in the control of the onset of puberty. To this end, female rats were i.c.v. injected with vehicle or AMPA (agonist of AMPA receptors: 0.1 or 0.5 nmol/day) between 26 and 30 days (Experiment 1), or 30 and 34 days (Experiment 2) of age. Serum concentrations of PRL, LH and estradiol were measured before drug administration, 10 min after the last injection, at vaginal opening (VO) and at first estrus (FE) presentation. In both experiments, AMPA administration inhibited PRL and estradiol secretion without affecting LH release. When AMPA was administered between 26 and 30 days a significant delay in the day of vaginal opening was observed. These results confirmed the inhibitory effect of AMPA on PRL secretion and suggests a role of AMPA receptors in the control of puberty onset.     

Effect of age at weaning on the physiological stress response and temperament of two beef cattle breeds

The study was conducted to evaluate the effect of age at weaning and breed on the stress response of calves to weaning and their temperament. At calving, 14 Parda de Montaña calves and 14 Pirenaica calves were randomly assigned to either early weaning (at 90 days) or traditional weaning (at 150 days) treatment. During nursing, calves were allowed to suckle their dams twice a day for 30 min. After weaning, calves were placed in an adjacent barn without access to their dams, where they remained for 7 days. On day 8 after weaning, they were transported to a feedlot where they received an intensive diet. Blood samples were taken 168 h before weaning (baseline) and 6, 24, 48 and 168 h after weaning for cortisol, fibrinogen and haematology analyses, and temperament was measured 90 and 180 days after weaning with the flight speed test. Cortisol concentration increased after weaning, irrespective of age at weaning. Early-weaned calves had a lower fibrinogen baseline and a greater increase in fibrinogen concentrations 48 h after weaning than traditionally weaned calves. Moreover, fibrinogen concentration returned to baseline values 168 h after weaning in traditionally weaned calves, whereas it remained high in early-weaned calves. Concerning breed effects, Pirenaica calves had higher cortisol concentration and fibrinogen increments after weaning than Parda de Montaña calves. Slight alterations occurred after weaning in haematology, but all parameters returned to baseline values 168 h after weaning, with no significant effects of age at weaning or breed. Despite the absence of clinical signs, early-weaned calves of both breeds suffered marginal anaemia, according to haemoglobin values. Regardless of age at weaning, Pirenaica calves had greater reactivity to human presence than Parda de Montaña calves, according to their higher flight speed values measured. Finally, early-weaned calves were lighter than traditionally weaned calves at weaning, but had similar weight gains in the feedlot. Consequently, they needed an additional 40 days to reach the target weight, irrespective of breed. Therefore, age at weaning had no major effects on the stress response to weaning or temperament, but early weaning increased the length of the feedlot period. On the other hand, Pirenaica calves were more reactive than Parda de Montaña calves to the stress of weaning and human presence.     

Anovulation in adult female rats after neonatal intracerebral implantation of oestrogen.

Four-day-old female rats were bilaterally implanted with paraffin micropellets containing 0.5% oestradiol benzoate (OB) into the mediobasal hypothalamus or the corticomedial amygdala. Controls received intracerebral paraffin pellets or two s.c. OB-paraffin implants. None of the treatments influenced the date of vaginal opening and vaginal cyclicity at 50 days of age. Permanent vaginal oestrus between 100 and 116 days of age and anovulatory ovaries at autopsy on day 116 were found in rats that had been implanted with OB into the mediobasal hypothalamus, but not in the remaining animals. The findings demonstrate that the delayed anovulatory syndrome can be induced in female rats by the neonatal intrahypothalamic implantation of a very low dose of OB, and that the corticomedial amygdala seems not to be a site of oestrogen action in this sterilizing effect.     

Relationship between opposite changes of vaginal and vestibular impedance during estrous cycle in sows

Several bioimpedance techniques have been developed for noninvasive monitoring of reproductive events occurring in cyclic gilts and sows. Our objective was to compare the changes of vaginal and vestibular impedance during the porcine estrous cycle (experiment 1). In addition, we examined the causes of impedance variations in the vaginal vestibule during periestrus (experiment 2). The vaginal and vestibular impedance were measured with specially designed instruments. Sows were monitored for estrus via exposure to a sexually mature boar. The impedance in the vagina decreased gradually after weaning (P<0.01) reaching its nadir 2 days before estrus and increased during estrus to near maximum 2 days after estrus onset (P<0.01). The vaginal impedance during diestrus reached approximately the same level as on the weaning day. In contrast, the impedance in the vaginal vestibule increased gradually after weaning, then markedly during estrus (P<0.01) reaching its maximum 2 days after the onset of estrus followed by an abrupt decrease during early diestrus (P<0.01). The vestibular impedance after early diestrus reached almost the same level as before estrus. A significant negative correlation was found between the vaginal impedance in proestrus and vestibular impedance in periestrus. In experiment 2, interaction of the interval from weaning to estrus and parity significantly influenced the vestibular impedance in periestrus. The breed of sows did not affect the impedance in the vaginal vestibule through the whole experiment. From the present study we conclude that closely related inverse changes of the vaginal and vestibular impedance take place in pigs during the estrous cycle.     

Nutritional influences on sexual maturation in the rat

The effect of altered nutrition on sexual maturation may depend in part on the nature and timing of the dietary change. The data are conflicting as to whether rats undernourished before weaning but normally fed after weaning have delayed puberty, but such undernourished rats clearly weigh less at vaginal opening than do normally fed animals. Altered nutrition after weaning can change the timing of puberty, and in such cases the body weight at puberty of the animals given the modified diet is frequently abnormal. The factors regulating the age and weight at puberty of rats fed altered diets seem to include the degree of underfeeding, as reflected in the growth rate, and the composition of the diet. Undernourished immature male rats have low serum testosterone secondary to gonadotropin deficiency. Basal luteinizing hormone (LH) in these animals is either low or "inappropriately normal" relative to their hypoandrogenic state (low serum testosterone and sexual accessory gland weights), and serum LH increases after luteinizing hormone-releasing hormone (LHRH) or castration are normal or minimally reduced. Serum follicle-stimulating hormone (FSH) in undernourished rats is subnormal basally and after administration of LHRH, but not after castration, which suggests that the low basal serum FSH is due to inhibition of FSH output by a testicular factor. Spermatogenesis may be unaltered by dietary changes severe enough to cause hypoandrogenism, although very severe under-nutrition will impair sperm production.     

Body composition at vaginal opening in mice as influenced by food intake and photoperiod: tests of critical body weight and composition hypotheses for puberty onset

To determine if photoperiod and food intake influenced age and body composition at vaginal opening (VO), 80 mice were exposed to either 18L:6D (LL) or 6L:18D (SL) and fed either 3.64 g/mouse per day (HIGH) or 2.73 g/mouse per day (LOW) of Wayne Lab-Blox. Treatments began at weaning (21 to 23 days of age). Mice were killed at VO and analyzed for fat, water and protein content. Mice fed the HIGH ration were younger (P less than 0.01; 35.9 vs. 41.1 days) and heavier (P less than 0.001; 17.8 vs. 15.4 g) at VO than mice fed the LOW ration. Neither age nor weight at VO were affected by photoperiod. Fat content at VO was greater (P less than 0.001) for HIGH than LOW. Differences in fat percentage at VO between diet groups were greater for LL (9.7 vs. 7.0%) than for SL (9.1 vs. 8.6%), resulting in a photoperiod X ration interaction (P less than 0.01). Percent water and protein at VO were not significantly affected by photoperiod or diet. Within-group correlations between body weight and age at VO were not significant. Body water/body weight was inversely proportional to age at VO. This resulted from increased fat deposition in older animals of the LL-LOW group and increased percent protein in older animals of other groups. We concluded that dietary intake did not influence age at VO by affecting age at which a critical body weight or fat content were attained. A photoperiod of 18L:6D did not affect age at VO relative to 6L:18D.