Postpartum aggression in mice: Experiential and environmental factors

Six experiments were conducted to assess the influence of duration of lactation, the presence of young, and the stimulus characteristics of intruder animals upon postpartum aggression of mice. The first experiment showed that postpartum aggression toward conspecifics was highest between Day 3 and Day 8, declined between Day 9 and Day 14, and was present toward males but absent toward females between Day 15 and Day 21 of the lactation period. Experiment 2 showed that lactating mice rarely attacked conspecifics to which they had been previously exposed but would readily attack strangers. Experiment 3 and 4 demonstrated that lactating animals never attacked intruders when tested 5 hr after pup removal. However, placement of young behind a wire partition in the home-cage for 5 hr or replacement of the offspring for as little as 5 min following 5 hr of separation restored postpartum aggression. The fifth experiment showed that 1- and 10-day old intruders were seldom attacked while intense aggression was directed against 14- and 20-day old intruders. Finally, Experiment 6 demonstrated that 14-day old intruders whose hair was removed were rarely attacked.     

Induction of maternal nest building in virgin female mice by the presentation of young

Ovariectomized and intact adult virgin female mice presented with two 1-day-old mouse pups built larger nests and more nests rated “maternal” than did intact animals not proffered young. Virgin females presented with live 1-day olds behind a wire partition in their homecages also constructed larger nests and more nests rated “maternal” as compared to animals presented with freshly killed 1-day olds, 19-day olds, or with no pups behind the partition.     

Cytosol and nuclear estrogen receptor binding in the preoptic area and hypothalamus of female rats during pregnancy and ovariectomized, nulliparous rats after steroid priming: correlation with maternal behavior

In a previous study, high nuclear estrogen receptor concentrations in the preoptic area (POA) were found on Day 16 of pregnancy to prime females to respond to a subsequent low dose of estradiol benzoate (EB) after hysterectomy-ovariectomy by exhibiting maternal behavior in 48 hr. Receptor concentrations in the POA were found to be higher than those in the hypothalamus (HYP). The present study investigated when nuclear estrogen receptors increase during pregnancy in POA and when the difference in receptor concentrations between POA and HYP occurs. An attempt was made to reproduce these pregnancy changes with a 16-day treatment of estrogen and progesterone in ovariectomized (OVX), nulliparous rats. In Experiment 1, we measured cytosol and nuclear estrogen receptor concentrations in the POA and HYP of female rats during pregnancy. Nuclear receptor concentrations in the POA increased beginning on Day 10, increased again on Day 16, and continued at this high level for the remainder of pregnancy. Nuclear estrogen receptor concentrations in the HYP remained at a lower level throughout most of pregnancy until Day 22 when they increased significantly. In Experiment 2, we tested the maternal behavior and measured estrogen receptor concentrations in OVX, steroid-primed, nulliparous rats after hysterectomy (H) and EB treatment. While 90% of estradiol (E) + progesterone (P)-primed females displayed short-latency maternal behavior 48 hr after H and EB treatment, 46% of E + vehicle (V)-treated controls were maternal. At 0 hr (prior to H and EB treatment), there was a significantly larger nuclear receptor accumulation in the POA but significantly attenuated receptor binding in the HYP. P treatment significantly affected cytosol and nuclear estrogen receptor dynamics. Differences in nuclear estrogen receptor concentrations were shown to be based on the number of available binding sites and not to changes in receptor affinity for estradiol.     

Prepartum onset of maternal behavior in hamsters and the effects of estrogen and progesterone.

The onset of maternal behavior in pregnant hamsters was measured by presenting foster pups at 0900 and 2100 hr on Day 15 and at 0300, 0500, and 0700 hr on Day 16 and then at hourly intervals until parturition began. The occurrence of parturition was determined at each maternal test and at 0.5 hr intervals beginning at 0700 hr on Day 16. Nulliparous and primiparous animals became maternal at approximately the same time on Day 16, 2 and 6 hr prepartum, respectively, demonstrating that parturition is not essential for maternal behavior. The second experiment showed that nulliparous females injected with either 1 μg or 10 μgm estradiol-17β (E₂), 0.1 mgm progesterone (P), 10 μgm E₂ plus 0.1 mgm P, or oil at 1200 hr on Day 15 became maternal at the same time of day (0800–1000 hr) while parturition was delayed 8 hr in females receiving P. The results suggest a dissociation between the regulation of parturition and maternal care and are compared to previous research into the hormonal basis of maternal behavior in rats.     

Parity-associated reductions in behavioral sensitivity to opiates

Behavioral and physiological responses differ between primiparous and multiparous female rodents. Specifically, multiparous females respond with the full repertoire of maternal behaviors much more rapidly and with greater intensity than their primiparous counterparts. Since opiates inhibit the expression of maternal behavior in postpartum rats and can be reversed by means of the opiate antagonist naloxone, we investigated whether multiparous females would be resistant to the inhibitory effects of opiates on maternal behavior, relative to primiparous females. In Experiment 1 we evaluated the effects of a range of doses of morphine sulfate (MS; 0.625, 1.25, 2.5, 5.0, and 10.0 mg/kg or saline) on maternal behavior in primiparous females on Days 5-6 of lactation. The 5.0 and 10.0 mg/kg doses effectively disrupted maternal behavior, whereas the lower doses were ineffective or only marginally disruptive. In Experiment 2, age-matched female rats were timed-mated and tested for maternal behavior from Day 5 to 13 of lactation, after daily injections of the 5.0 mg/kg dose of MS. On Day 5 of lactation, this morphine treatment eliminated full maternal behavior in 87% of the primiparous animals, but only 37% of the multiparous animals were affected. By Day 10 of lactation, 100% of the multiparous females displayed full maternal behavior after MS treatment, whereas only 69% of primiparous females were responsive. In Experiment 3, analgesic responses were measured both in rats experiencing their initial or second pregnancy, and in postpartum, lactating rats after MS (5.0 mg/kg) administration. Using a tail-flick apparatus to measure analgesia, we found multigravid females to be significantly less analgesic prepartum than primigravid females, suggesting less sensitivity to endogenous opioids.(ABSTRACT TRUNCATED AT 250 WORDS)     

Induction of maternal behaviors in primigravid rats by ovariectomy,hysterectomy, or ovariectomy plus hysterectomy: Effect of length of gestation

The effects of terminating pregnancy by means of ovariectomy (O), hysterectomy (H), or ovariectomy plus hysterectomy (OH) at different stages of gestation upon the latency to initiation of maternal behavior were examined in primigravid rats. O, H, or OH on either Day 13 or 17 of pregnancy resulted in an accelerated onset of maternal behaviors (median range = 1.0–2.0 day latency for O, H, or OH animals vs 4.0–5.0 day latency for sham-operated intacts). However, O and H on Day 8 of pregnancy were ineffective in inducing a rapid onset of maternal behavior, while OH on Day 8 of pregnancy only slightly facilitated the onset of maternal behavior when compared to animals sham-operated on Day 8 of pregnancy. O, H, and OH on either Day 13 or 17 of gestation were also significantly more effective in rapidly inducing maternal behaviors than the corresponding surgeries performed on Day 8 of pregnancy. These data suggest that the onset of maternal behavior in the pregnant rat can be rapidly induced after mid-pregnancy by surgical procedures that presumably result in a rapid decline in serum steroids of ovarian origin.     

Progesterone inhibition of estrogen-induced maternal behavior in hysterectomized-ovariectomized virgin rats.

Hysterectomized-ovariectomized virgin rats were tested for maternal behavior following treatment with 100 μg/kg EB immediately at surgery and either oil, 0.5 or 5.0 mg progesterone either 0, 24 or 44 hr following surgery. Stimulus pups were presented 48 hr postoperatively which is counted as Day 0 of testing. EB + oil-treated females displayed short-latency maternal behavior beginning on Day 0. The injection of 5.0 mg progesterone at 0, 24, or 44 hr significantly inhibited the onset of maternal care while the effect of the lower dose of progesterone depended upon the timing of its administration in relation to that of EB. At a dose of 0.5 mg, progesterone given 24 hr following EB, inhibited the appearance of maternal behavior but had no effect given at 44 hr, and resulted in only a partial delay when given at the same time as the EB. Possible mechanisms by which progesterone interfered with the display of maternal behavior were discussed.     

The estrogenic arousal of aggressive behavior in female mice

Experiments were conducted to determine the conditions under which estrogen would promote male-like aggressive behavior in female mice. The results of the first experiment showed that most females chronically exposed to testosterone propionate (TP) in adulthood fought, whereas females similarly treated with estradiol benzoate (EB) did not display aggression. Another experiment found that, when either TP or EB was administered on the day of birth, adult females displayed aggression in response to daily EB injections during adult life. Also, the potentiating effect of neonatal hormone exposure declined over the first 12 days postpartum, as 100% of the Day 0, 75% of the Day 6, and 0% of the Day 12 and 18 TP-treated females fought in response to daily injections of 40 μg of EB in adulthood. The final study showed that, under the test conditions employed, the failure of a chronic adult EB regimen to promote aggression was not due to a competing tendency to display female sexual behavior.     

Reproductive success,postpartum maternal behavior,and masculine sexual behavior of neonatally androgenized female hamsters

Sex differences in maternal behavior induced by pup stimulation (sensitization) have been reported for rats and hamsters and may be affected by the presence or absence of perinatal androgen treatment. Postpartum maternal behavior and litter survival in golden hamsters treated with testosterone propionate (TP) as neonates were studied. A high dose of TP (300 μg) eliminated feminine reproductive capacity when given on Day 2 or 4 postpartum and had no discernible effect on Day 12. Treatment on Days 6, 8, or 10 resulted in treatment day-dependent deficiencies in reproductive success which fell short of sterility in most females. These deficiencies included low birth weight, weight gain, and higher litter losses than controls. However, the maternal behavior of TP dams, as measured by retrieval and crouching, appeared to be normal. The disparity between delivery and successful rearing of normal-weight young may include uterine incompetence, lactation deficiency, and hypercannibalism. Behavioral masculinization was a more sensitive index of neonatal androgen action than any aspect of defeminization, but the two phenomena were dissociated in individuals.     

Growth hormone is secreted by ectopic pituitary grafts and stimulates maternal behavior in rats

The onset of maternal behavior at parturition in rats is hormonally regulated. Recently, we reported that treatment of behaviorally inexperienced, hypophysectomized (hypox), ovariectomized (ovx) rats with a sequential steroid treatment of progesterone (P) and estradiol (E2), and either ectopic anterior pituitary grafts or prolactin (PRL), stimulated maternal responsiveness toward foster young. That growth hormone (GH) has a number of PRL-like activities led us to ask whether the actions of PRL on maternal behavior were specific to PRL or might be shared by other PRL-like protein hormone, i.e., GH. In Experiment 1 we quantified plasma concentrations of GH and PRL by RIA in groups of hypox female rats that were ovariectomized and treated with a combination of ectopic pituitary grafts (Days 1-23) and Silastic capsules filled with P (Days 1-11) and E2 (Days 11-23). Blood samples were collected from Days 1 to 23 of treatment. Both plasma PRL and GH levels increased after grafting, initially rising 10- to 60-fold by Day 4 and gradually declining throughout the remainder of the 23-day sampling period. Throughout the 3-week period after grafting plasma GH levels were as high or higher than those of PRL. In Experiment 2 the behavioral effects of exogenously administered ovine (o)-GH were measured in groups of hypox, ovx rats that were treated with P and E2 as in Experiment 1. Experimental rats were injected twice daily with 0.25 mg oGH beginning on Day 1. Testing for maternal behavior toward foster young was conducted daily from Day 12 to Day 22. In steroid-treated rats, GH treatment stimulated a more rapid onset of maternal behavior (latencies of 3 vs greater than 10 days for vehicle-injected controls). These data indicate that GH, like PRL, is secreted by ectopic pituitary grafts and is capable of stimulating maternal behavior.     

Differential sensitivity of mounting and lordosis control systems to early androgen treatment in male and female hamsters.

The effects of early testosterone propionate (TP) treatment on the adult sexual behavior of hamsters were investigated in two experiments. In Expt. I, male and female pups were injected with oil vehicle or 1, 5, 10, 50, 100, or 250 μg of TP 24 hr after birth. In Expt. II, males and females received either oil or 10 μg of TP on the day of birth (Day 1), Day 3, Day 5, Day 7, or Day 9. At 70 days of age all animals were gonadectomized and 10 days later tested for lordosis behavior after estrogen and progesterone priming. One week after the test for female behavior all females began receiving 500 μg of TP each day and were tested for mounting and intromission behavior three times at 10 day intervals. Lordosis behavior was inhibited by as little as 5 μg of TP given 24 hr after birth. In males this dose produced the maximal effect, but in females increasing dosages resulted in a proportional decrease in lordosis duration. One μg of TP neonatally facilitated later mounting and intromission behavior in females and 250 μg of TP was no more effective than 1 μg. Lordosis duration was inhibited in females by 10 μg of TP on either Day 1 or 3, however, mounts and intromissions were facilitated by TP treatment on Day 1, 3, 5 or 7. These experiments demonstrate that the mechanisms mediating masculine behavior are more sensitive to neonatal TP treatment than are the mechanisms mediating lordosis behavior.     

Prenatal stress and maternal behavior in intact virgin rats: response latencies are decreased in males and increased in females

The repeated findings that levels of various male-typical behaviors (e.g., copulatory behavior and intermale aggression) are reduced in prenatally stressed (P-S) males, coupled with reports of effects on female physiology and behavior, prompted us to examine the maternal behaviors of P-S animals toward young. Sprague-Dawley female rats were timed-mated (+ sperm = Day 1). From Gestation Days 15 to 22 experimental females were subjected to heat and restraint stress. Control females remained undisturbed throughout pregnancy. The offspring, as adults, were assessed for maternal behavior. P-S males exhibited a significantly shorter latency (in days) to show full maternal behavior (FMB) than Control males, median = 5.0 vs 8.0, respectively. P-S females, on the other hand, exhibited a significantly longer latency than Control females to show FMB (7.0 vs 3.0, respectively). as well as longer latencies to retrieve one, two, or three pups, to begin to crouch over pups, and to build nests in response to young. Sex differences were apparent between Control males and Control females (females were more responsive to young). In contrast, P-S males and Control females exhibited similar latencies to show components of FMB (3-5 days), as did P-S females and Control males (7-9 days). These data demonstrate, therefore, that prenatal stress eliminates the sex difference normally observed in pup-induced maternal behavior. Moreover, the data suggest that prenatal stress renders the male's responsiveness to young more "female-like," while conversely rendering the response of the female more "male-like."     

Mammary stimulation and maternal aggression in rodents: thelectomy fails to reduce pre- or postpartum aggression in rats

In mice, tactile stimulation of the nipples appears to be critical for the onset of postpartum maternal aggression. Surgical removal of the nipples (thelectomy) blocks aggression if performed prior to parturition. In rats, indirect evidence suggests a similar role for nipple stimulation in maternal aggression. Two experiments were undertaken to determine whether thelectomy prior to mating reduces pregnancy-induced and/or postpartum aggression in this species. In the first, thelectomized and sham-thelectomized females were subjected to home cage tests (pups, if any, present) with unfamiliar male intruders on Gestation Days 18 and 21 and Lactation Days 3 and 5. Additional groups of thelectomized females were tested one time only on either Lactation Day 5 or 12. Thelectomized and control females were equally aggressive; postpartum, nearly all females in both groups attacked. Experiment 2 used females that were hysterectomized-ovariectomized (HO) on Gestation Day 16. Such females are not aggressive prior to initiating maternal behavior, but become highly aggressive (over 80% attacking) after commencing maternal care. Females again were thelectomized or sham-thelectomized prior to mating. On Day 16 HO was performed, and 48 hr later continuous exposure to pups was begun. After the females had displayed maternal behavior for 1.5-2 days, intruder tests were conducted. All females attacked at least once, with no differences between treatment groups. Thus thelectomy does not reduce maternal aggression in the rat. This finding, however, does not preclude a role for tactile ventral stimulation in mediating maternal aggression.     

Hormonal basis of hysterectomy-induced maternal behavior during pregnancy in the rat.

Hysterectomy during the last half of pregnancy (i.e., Day 10–19) induces a rapid onset of maternal behavior; ovariectomy in addition to hysterectomy, prevents this effect. Estradiol and progesterone were tested for their ability to restore short-latency maternal behavior in hysterectomized-ovariectomized (HO) females operated on the 10th, 13th, 16th and 19th days of pregnancy. A single injection of either 20 μg/kg or 100 μg/kg estradiol benzoate (EB) immediately following HO either alone or followed by 0.5 mg progesterone (P) 44 hr later restored short-latency maternal behavior similar to that observed following hysterectomy only. The lower dose of EB was found to be equally effective at all stages of pregnancy and P was unnecessary to induce maternal behavior. The effectiveness of EB in inducing maternal behavior was discussed in relation to the hormonal changes which follow hysterectomy during pregnancy and to those which are associated with the normal onset of maternal behavior around parturition.     

Pregnancy termination by prostaglandin F2α stimulates maternal behavior in the rat

Treatment with PGF_2α resulted in the termination of pregnancy in 16- and 19-day pregnant rats, but not in 10- or 13-day pregnant rats. Rats that aborted displayed a rapid onset of maternal behavior when tested with foster pups. Aborted rats also displayed sexual receptivity and ovulation: these phenomena resemble the sequence of events following hysterectomy on the same days of pregnancy. Both can be related to the events surrounding normal parturition in the rat. The results are interpreted as due to a pregnancy-induced deactivation of the factor in the uterus that prevents estrogen from stimulating maternal behavior in nonpregnant females. In the absence of this factor, the PGF_2α-induced rise in estrogen secretion facilitates maternal behavior and sexual behavior and induces ovulation.     

The role of the young in the control of the hormonal events during lactation and behavioral weaning in the golden hamster

This study was designed to determine the relationship between the behavioral and physiological changes occurring in the hamster over the course of lactation. Experimental (E) females were given litters of six 3-day-old pups on Day 13 of lactation and allowed to raise them. Control (C) females raised their own litters of six pups. Groups of E and C females were decapitated at 1000 and 1700 hr when their pups were either 8, 18, or 28 days old. Nursing was observed for 1 hr on the 3 days prior to autopsy. Nursing behavior disappeared by Day 28 in C females. E females exhibited nursing behavior at levels equal to those observed in C females until E pups were 28 days old and 38 days had elapsed since parturition. Despite the fact that E mothers continued to nurse pups on day 18 postpartum when pups were 8 days old, E pups showed lower growth rates than did C pups. Prolactin (PRL) levels remained elevated when E pups were 8 days old even though E pups did not grow normally. PRL levels decreased over time in both C and E females and reached baseline by Day 28 although nursing behavior was still elevated in E females. Thus, nursing behavior did not stimulate PRL release during late lactation. Estradiol (E₂) levels in C females remained at baseline until Day 28 when levels increased. LH, FSH, and Progesterone (P) levels in C females showed a dramatic diurnal pattern which disappeared by Day 28, when levels dropped. Levels of E₂, P, LH, and FSH in E females closely paralleled those of C females only when groups were compared with respect to pup age. Thus, behavioral weaning and the return to estrous cyclicity appear to be dependent upon the age of the pups rather than the time elapsed since parturition. However, milk production and PRL release appear to be more closely tied to the number of days postpartum and can be dissociated from the amount of nursing behavior observed.     

Changes in serum immunoreactive inhibin and follicle-stimulating hormone during gonadal development in male and female rats.

We have measured serum and ovarian immunoreactive inhibin alpha (irI alpha) and serum FSH in fetal and neonatal rats from 20 days of gestation until 40 days of age. For animals aged 10 days or older, serum measurements were made on intact and gonadectomized animals. Serum irI alpha was detectable in intact male and female rats at all ages studied. In females, irI alpha levels were low until Day 5 and then increased steadily to peak at Day 25. Thereafter they declined until Day 35 to reach levels typical of adult females. There was a significant decrease in irI alpha levels 24 h after ovariectomy at all ages. Serum FSH levels in females were low until Day 7, then increased rapidly to plateau from Days 10-15. The levels then declined until Day 25 and were generally unchanged after that time. There was a significant increase in FSH 24 h after ovariectomy in rats aged 20 days and older, and in younger rats by 48 h after ovariectomy. In male rats, serum irI alpha levels were significantly higher than females until Day 7. The levels increased at Day 7 and then remained relatively constant until Day 20, after which they declined to reach typical adult male levels. Serum irI alpha levels were significantly lower in males than females from Days 25-40. There was a significant decrease in serum irI alpha 24 h after castration at all ages studied. Serum FSH levels in males were low until Day 20, increased at Day 25, and thereafter remained relatively unchanged.(ABSTRACT TRUNCATED AT 250 WORDS)     

Progesterone inhibition of maternal behavior in the rat

The present series of experiments investigated the role of progesterone in inhibiting the onset of maternal behavior in the rat. Female rats hysterectomized and ovariectomized on Day 16 of pregnancy and injected subcutaneously with 20 μg/kg of estradiol benzoate (EB) show a short latency to onset of maternal behavior when presented with test pups 48 hr later. A subcutaneous injection of either 1 or 5 mg of progesterone on Day 16 of pregnancy and again 24 hr later inhibited this EB-induced short-latency onset of maternal behavior. The central neural site at which progesterone might act to produce this inhibitory effect was explored. Famale rats, hysterectomized and ovariectomized on Day 16 of pregnancy and injected subcutaneously with EB, received implants of crystalline progesterone on Day 16 of pregnancy into either the medial preoptic area, ventromedial hypothalamus, midbrain tegmentum, dorsal raphe nucleus, or median raphe nucleus. No inhibitory effects were found and all females showed a short-latency onset of maternal behavior. Several possible explanations for this lack of inhibitory effect of intracerebral implantation of progesterone are discussed.     

Masculine sexual behavior in male and female rats following perinatal manipulation of androgen: Effects of genital anesthetization and sexual experience

Androgenized females, Day 1 male castrates, normal males, and normal females were tested for mounting behavior as adults following TP administration (1 mg/day). Genital anesthetization was used to eliminate all intromission and ejaculation behavior. Results showed that sexually-naive normal males and androgenized females mounted significantly more then Day 1 male castrates, while the Day 1 male castrates mounted significantly more than normal females. Sexually-experienced normal males and androgenized females displayed a significant facilitation of mounting behavior as compared to the sexually-naive animals. Tests of masculine copulatory behavior without genital anesthetization also were conducted with androgenized females and normal males. In these tests, androgenized females were indistinguishable from normal males in all aspects of the complete masculine pattern. The present results provide evidence that during perinatal development androgen acts directly on neural systems which will later regulate adult masculine sexual behavior.     

Effects of environmental stress or ACTH treatment during pregnancy on maternal and fetal plasma androstenedione in the rat

Prenatal stress applied during the last trimester of pregnancy has been shown to alter fetal development and influence adult sexual behavior. Since androstenedione (Δ⁴) has the potential to participate in differentiation processes, this study was designed to assess the effect of prenatal stress on maternal and fetal Δ⁴ titers. Restraint/illumination/heat (environmental stress) or ACTH injections were used to stress pregnant rat dams beginning on Day 14 of pregnancy. Blood samples and organ weights were obtained from nonpregnant animals, pregnant rats on Days 5, 10, 15, 18, and 20 of pregnancy, and fetuses on Days 18 and 20 of gestation. Maternal and male and female fetal Δ⁴ titers were determined by radioimmunoassay. ACTH and environmental stress significantly reduced fetal body weight and male anogenital distance. Environmental stress also significantly reduced the size of 20-day fetal adrenals and testes. Each treatment caused significant short-term (1 hr after treatment) and long-term (16 hr after treatment) elevation of maternal plasma Δ⁴ on Days 15 and 18 of gestation, but only short-term elevation of Δ⁴ titers on Day 20. ACTH treatment did not cause long-term elevation of fetal Δ⁴ although both ACTH treatment and environmental stress generated a significant short-term increase in fetal Δ⁴ titers. Environmental stress produced long-term elevation of fetal Δ⁴ in 18-day fetuses of both sexes and in 20-day female fetuses. It is concluded that maternal stress and exogenous ACTH significantly elevate maternal and fetal Δ⁴ titers during the prenatal period postulated to be critical in sexual differentiation of the rat brain.