Pup Exposure Research Articles

Attenuation of maternal behavior in virgin CD-1 mice by methylphenidate hydrochloride

The administration of methylphenidate hydrochloride (MPH) to girls and women has increased in the last decade and the potential for mothers to receive this medication has also increased. Because substances that alter the dopaminergic systems can also disrupt maternal behavior, and MPH acts on dopaminergic neurons, we evaluated the influence of MPH on maternal behavior. The maternal induction paradigm allowed us to assess changes in spontaneous maternal behavior as a result of repeated exposure to MPH without exposing pups to the drug. Virgin female CD-1 mice received MPH (5 mg/kg) or saline daily, starting 3 days before pup exposure, and for the duration of the 10-day test period. Naïve groups of three pups were placed with the female each day and maternal behavior was assessed during 10-minute observation periods 1 h post-injection. MPH-treated females showed significantly less maternal behavior, including reduced pup licking and crouching over pups, compared to saline treated females. MPH-treated females also exhibited higher activity levels than saline treated females. Given the disruption in spontaneous maternal behavior of MPH-treated mice, further research examining the relationship between maternal behavior and MPH exposure is warranted.

Predicting Maternal Rat and Pup Exposures: How Different are They?

Risk and safety assessments for early life exposures to environmental chemicals or pharmaceuticals based on cross-species extrapolation would greatly benefit from information on chemical dosimetry in the young. Although relevant toxicity studies involve exposures during multiple life stages, the mother's exposure dose is frequently used for extrapolation of rodent toxicity findings to humans and represents a substantial source of uncertainty. A compartmental pharmacokinetic model augmented with biological information on factors changing during lactation and early postweaning was developed. The model uses adult pharmacokinetics, milk distribution, and relevant postnatal biology to predict dosimetry in the young for chemicals. The model addressed three dosing strategies employed in toxicity studies (gavage, constant ppm diet, and adjusted ppm diet) and the impact of different pharmacokinetic properties such as rates of clearance, milk distribution, and volume of distribution on the pup exposure doses and internal dosimetry. Developmental delays in clearance and recirculation of chemical in excreta from the pup to mother were evaluated. Following comparison with data for two chemicals, predictions were made for theoretical chemicals with a range of characteristics. Pup exposure was generally lower than the mother's with a shorter half-life, lower milk transfer, larger volume of distribution, and gavage dosing, while higher with longer half-life, higher milk transfer, smaller volume of distribution, and dietary exposures. The present model demonstrated pup exposures do not always parallel the mother's. The model predictions can be used to help design early life toxicity and pharmacokinetic studies and better interpret study findings.

Reproductive experience alters hippocampal neurogenesis during the postpartum period in the dam

Pregnancy and the postpartum period are a time of maximal neural and behavioral plasticity. Recent work has shown that hippocampus-dependent learning and memory performance and hippocampus morphology are affected by motherhood and reproductive experience (number of times pregnant and given birth). Adult neurogenesis in the dentate gyrus of the hippocampus is influenced by steroid hormones such as estradiol and corticosterone, which fluctuate during pregnancy and the postpartum period. Thus, it is possible that hippocampal neurogenesis may be affected by motherhood and reproductive experience. The present study aimed to investigate the role of reproductive experience on hippocampal neurogenesis via cell proliferation and cell survival and to determine whether differences were due to the effect of pregnancy and/or pup-exposure alone. Four groups of female Sprague–Dawley rats were used; multiparous, primiparous, nulliparous, and nulliparous rats exposed to pups. All rats were injected with 5-bromo-2-deoxyuridine (BrdU) (200 mg/kg) approximately 24 h after birth/pup-exposure with age-matched controls. Rats were perfused either 24 h (Expt. 1: Cell proliferation) or 21 days (Expt. 2: Cell survival) after BrdU injection. Results show there is a significant decrease in cell proliferation in the dentate gyrus of primiparous and multiparous rats during the early postpartum period, and a decrease in cell survival in the dentate gyrus during the postpartum in primiparous rats, regardless of pup-exposure, compared with all other groups. In addition, brief pup exposure to nulliparous rats significantly increased cell proliferation and cell death in the dentate gyrus, while 22 days of pup exposure to nulliparous rats (sensitized rats) resulted in increased cell survival and cell death in the dentate gyrus. Collectively these results indicate that reproductive experience significantly affects hippocampal neurogenesis and that these effects are not due to the effect of pregnancy or pup-exposure alone.

ERK-FosB signaling in dorsal MPOA neurons plays a major role in the initiation of parental behavior in mice

During mouse parental behavior, neurons in the dorsal medial preoptic area (MPOAd) are activated and express transcription factors such as c-Fos and FosB. FosB-knockout mice are reported to be defective in parental care. To clarify molecular signaling responsible for parental behavior, we investigated gene expression profiles in the MPOAd of parental versus nonparental mice. We identified upregulation of NGFI-B, SPRY1, and Rad in parental mice, together with c-Fos and FosB. A common inducer of these genes, the extracellular signal regulated kinase (ERK) was phosphorylated in MPOAd neurons upon pup exposure. Pharmacological blockade of ERK phosphorylation inhibited the FosB upregulation in MPOAd, and the initiation of pup retrieving in virgin female mice, but did not affect the established parenting in parous females. Furthermore, induction of SPRY1 and Rad was impaired in MPOAd of nonparental FosB-knockout mice. These results suggest the pivotal role of ERK-FosB signaling in the initiation of parental care.

Mother rabbits and their offspring: Timing is everything

Mother rabbits nurse once every 24 hr for ca. 3 min. The contribution of pup stimulation to the regulation of this process was investigated by: reducing litter size to one pup, covering all the mother's nipples, or taping the mouths of an eight-pup litter. Time inside the nest box with the young significantly increased under all these conditions. Yet, eight pups sucking on a single nipple allowed a normal time in the nest box. Bringing the litter to the mother earlier than usual promoted maternal behavior expression twice in 24 hr; the proportion of responsive females and the time at which they responded depended on the size of the litter suckled. These findings indicate that: (a) suckling determines the time inside the nest box and prevents mothers from responding to further pup stimulation for a variable time; (b) pup exposure promotes maternal behavior, even "out of phase" with circadian rhythmicity.

Species and individual differences in juvenile female alloparental care are associated with oxytocin receptor density in the striatum and the lateral septum

The neuropeptide oxytocin has been implicated in the regulation of affiliative behavior and maternal responsiveness in several mammalian species. Rodent species vary considerably in the expression of juvenile alloparental behavior. For example, alloparental behavior is spontaneous in juvenile female prairie voles (∼20 days of age), takes 1–3 days of pup exposure to develop in juvenile rats, and is nearly absent in juvenile mice and meadow voles. Here, we tested the hypothesis that species differences in pup responsiveness in juvenile rodents are associated with oxytocin receptor (OTR) density in specific brain regions. We found that OTR density in the nucleus accumbens (NA) is highest in juvenile prairie voles, intermediate in juvenile rats, and lowest in juvenile mice and meadow voles. In the caudate putamen (CP), OTR binding was highest in prairie voles, intermediate in rats and meadow voles, and lowest in mice. In contrast, the lateral septum (LS) shows an opposite pattern, with OTR binding being high in mice and meadow voles and low in prairie voles and rats. Thus, alloparental responsiveness in juvenile rodents is positively correlated with OTR density in the NA and CP and negatively correlated with OTR density in the LS. We then investigated whether a similar receptor–behavior relationship exists among juvenile female prairie voles by correlating individual variation in alloparental behavior with variation in OTR density. The time spent adopting crouching postures, the most distinctive component of alloparental behavior in juveniles, was positively correlated with OTR density in the NA ( r = 0.47) and CP ( r = 0.45) and negatively correlated with OTR density in the lateral septum ( r = −0.53). Thus, variation in OTR density in the NA, CP, and LS may underlie both species and individual differences in alloparental care in rodents.

Reproductive experience and activation of maternal memory.

The maternal and neurobiological responses of biological mothers and pup-induced maternal virgin rats were compared 55 and 80 days after an initial 2-day maternal experience. When tested for home cage responsiveness after prolonged isolation from young, the biological, primiparous rats displayed shorter maternal latencies. Primiparous females tested in the presence of pups on the elevated plus-maze displayed increased exploration of the open arms and increased c-Fos expression in the cortical nucleus of the amygdala. Pup exposure and parity also enhanced activation of the nucleus accumbens shell and medial nucleus of the amygdala, respectively. Therefore, although both nulliparous and primiparous rats retain a maternal memory for a prolonged time, the memory and neurochemical response appear stronger in primiparous mothers.

Developmental Toxicology Evaluations—Issues with Including Neurotoxicology and Immunotoxicology Assessments in Reproductive Toxicology Studies

Developmental and reproductive toxicology (DART) has routinely been a part of safety assessment. Attention is now focused on the effects of chemicals on the developing nervous and immune systems. This focus on developmental neurotoxicology (DNT) and developmental immunotoxicology (DIT) is based on the premise that children differ from adults in some aspects of their biology and, thus, may also differ in their responses to chemicals. This session's objective was to discuss issues common to DNT and DIT as they relate to DART protocols, including high dose selection and maternal toxicity, adequacy of pup exposure during lactation, use of a different dosing paradigm for DART versus DNT or DIT studies, and whether DIT and DNT endpoints can be incorporated into a single DART study for hazard identification purposes. Consensus was achieved on all topics except the adequacy for risk assessment purposes of the use of a limited number of endpoints for DIT and DNT, with the DNT endpoints being the primary focus of disagreement. Panelists indicated that a combination study design for hazard identification was feasible, though flexibility to meet the scientific needs of the project was emphasized. The adequacy of existing triggers for additional developmental studies was also questioned. Panelists iterated the importance of understanding pup exposure during the various life stages and the use of toxicokinetic data in designing these studies. The group agreed to consider the HESI ACSA Life Stages Task Force recommendations as a next step to address some of the issues and challenges raised during this session.

Pup exposure differentially enhances foraging ability in primiparous and nulliparous rats

The role of maternal experience (i.e., pregnancy and pup exposure) on rats' performance in a foraging task was assessed. Primiparous (P) and nulliparous (N) animals were either exposed to pups for 21 days (+) or received no pup exposure (−). Following habituation trials, all animals were tested in spatial and cued versions of the dry land maze (DLM) for three days (three trials per day). In the spatial DLM, the presence of pups decreased latencies in both N and P groups in Trial 5 and P+ rats exhibited shorter latencies to baited food wells than P− animals on Trial 6. In the subsequent probe trial, P+ animals spent significantly more time in proximity to the previously baited well than P− rats. Pups enhanced performance of both P+ and N+ groups in trial 6 of the cued test. Thus, in the spatial task, the individual components of the maternal experience (e.g., pregnancy, parturition, lactation, and pup exposure) converge to produce behavioral modifications in the DLM spatial and probe tasks that enable the female to care for her offspring, in this case, by enhancing foraging behavior. Further, in one trial of each version of the task, pup exposure enhanced performance in N animals suggesting that, in isolation, pup exposure may be a more important influence on ancillary maternal behavior than the pregnancy itself.

Variability in “spontaneous” maternal behavior is associated with anxiety-like behavior and affiliation in Naïve juvenile and adult female prairie voles (Microtus ochrogaster)

Juvenile female prairie voles (Microtus ochrogaster) are spontaneously maternal, while virgin adult females show significant variability in their response to first pup exposure, ranging from infanticidal to full maternal behavior. In the present study, we investigated whether differences in anxiety-like behavior and affiliation are associated with juvenile-adult and adult individual differences in the response to pups. Forty juvenile (19-20 days) and 42 adult (60-90 days) female prairie voles were exposed to pups for the first time and tested for maternal behavior, anxiety-like behavior (elevated plus maze, open field), and affiliation toward age-matched, same sex conspecifics. Juveniles displayed less anxiety-like behavior, were more affiliative to unfamiliar conspecifics, and interacted with pups more positively than adults. Adults that displayed maternal behavior spent less time immobile, made more crosses through the center of the open field arena, and were more affiliative than adults that attacked the pups. This suggests that lower locomotion or exploration in a novel environment and poor affiliative behavior are negatively associated with maternal responsiveness in female prairie voles.

Cyclic AMP response element-binding protein is required for normal maternal nurturing behavior

Analysis of mice with targeted disruptions of fosB or the gene encoding dopamine β-hydroxylase suggests that FosB and adrenergic signaling play critical roles in maternal nurturing behavior. The majority of neonates born to null females from either mutation fail to thrive, and virgin mutant females of both lines exhibit impaired pup retrieval. Considering whether FosB and adrenergic signaling might share a signaling pathway important for maternal behavior, we examined the role of a potential intermediary, cyclic AMP response element-binding protein (CREB). Here we report that ∼40% of neonates (all heterozygous) born to mice lacking the major isoforms of CREB ( Creb-αΔ −/−) died within several days of birth. In contrast, heterozygotes born to Creb-αΔ +/− females thrived. Cross-fostering demonstrated that neonates born to Creb-αΔ −/†/− females thrived when reared by wild-type females, and that Creb-αΔ −/− females were capable of rearing neonates whose maternal care was initiated by wild-type females. Further, virgin Creb-αΔ −/− females were deficient in pup retrieval despite exhibiting normal investigation of pups and of novel objects. No maternal behavior phenotype was present in mice with a null mutation of the cyclic AMP response element modulator ( Crem) gene. Interestingly, the number of cells immunostaining for phospho-CREB (on Ser 133) in the medial preoptic area of the hypothalamus, a key region for the expression of maternal behavior, increased nearly three-fold in wild-type mice following exposure to pups but not to novel objects. On the other hand, basal expression and induction of FosB in response to pup exposure appeared to be independent of CREB because levels were equivalent between wild-type and Creb-αΔ −/− females. These results implicate CREB in maternal nurturing behavior and suggest that CREB is not critical for expression or induction of FosB in adult virgin female mice.

Heterotrimeric G proteins of the Gq/11 family are crucial for the induction of maternal behavior in mice.

Heterotrimeric G proteins of the G(q/11) family transduce signals from a variety of neurotransmitter receptors and have therefore been implicated in several functions of the central nervous system. To investigate the potential role of G(q/11) signaling in behavior, we generated mice which lack the alpha-subunits of the two main members of the G(q/11) family, Galpha(q) and Galpha(11), selectively in the forebrain. We show here that forebrain Galpha(q/11)-deficient females do not display any maternal behavior such as nest building, pup retrieving, crouching, or nursing. However, olfaction, motor behavior and mammary gland function are normal in forebrain Galpha(q/11)-deficient females. We used c-fos immunohistochemistry to investigate pup-induced neuronal activation in different forebrain regions and found a significant reduction in the medial preoptic area, the bed nucleus of stria terminalis, and the lateral septum both in postpartum females and in virgin females after foster pup exposure. Pituitary function, especially prolactin release, was normal in forebrain Galpha(q/11)-deficient females, and activation of oxytocin receptor-positive neurons in the hypothalamus did not differ between genotypes. Our findings show that G(q/11) signaling is indispensable to the neuronal circuit that connects the perception of pup-related stimuli to the initiation of maternal behavior and that this defect cannot be attributed to either reduced systemic prolactin levels or impaired activation of oxytocin receptor-positive neurons of the hypothalamus.

The content of dopamine, serotonin, and their metabolites in the neural circuit that mediates maternal behavior in juvenile and adult rats

Continuous exposure of non-parturient rats to pups can induce maternal behavior similar in most aspects to that found in the postpartum rat. Surprisingly, young juvenile rats (20–24 days of age) only require 1–3 days of exposure to pups, while adults require 4–8 days before maternal behavior emerges. Dopamine (DA) and possibly serotonin (5-HT) may mediate the expression of adult maternal behavior. We hypothesize that postnatal changes in DA and 5-HT within the neural circuit that supports maternal behavior including the medial preoptic area (MPOA), medial and cortical amygdala (MCA), and nucleus accumbens (NAC), may underlie these differences in responsiveness across juveniles and adults. We measured DA, 5-HT, and their metabolites in postmortem samples of these regions in maternal and non-maternal juvenile and adult females. The only difference found across behavioral groups was that the MPOA of adults induced into maternal behavior by pup exposure had more DA than did that of isolated adult females or maternal juveniles. However, when adults versus juveniles were compared, the content of DA and 3,4-dihydroxyphenylacetic (DOPAC) was higher in the adult than in the juvenile NAC and MCA; the content of 5-HT and 5-hydroxyindoleacetic acid (5-HIAA) in these structures did not vary across the age groups. In contrast, higher levels of 5-HT and 5-HIAA were found in the MPOA in juveniles compared to adults. We propose that these region-specific age differences in DA and 5HT may underlie differences in juvenile–adult responses to pups.

Removal of the accessory olfactory bulbs promotes maternal behavior in virgin rabbits

Virgin rabbits exposed to foster pups for 14 days did not show maternal responsiveness. However, surgical removal of the accessory olfactory bulbs (AOB) activated maternal responsiveness (crouching over the litter inside the nest box for about 3 min, which is the normal duration of a nursing bout) in 37% of virgin rabbits ( P<0.007). This behavior appeared abruptly and was first observed between days 3 and 13 of pup exposure. This variation in the latency to respond maternally was not related with the number of sniffings or entrances into the nest box displayed by a female on the days that preceded crouching over the litter. Maternal responsiveness was not observed in any AOB-lesioned animals that were also ovariectomized ( P<0.02 versus AOB-lesioned with ovaries). These results indicate a tonic inhibitory action of the AOB over the expression of maternal behavior in virgin rabbits and a stimulation of maternal responsiveness by ovarian hormones following AOB lesions.

Single or multiple reproductive experiences attenuate neurobehavioral stress and fear responses in the female rat

The female brain is a dynamic structure, which expresses its plasticity most readily following reproductive experience (RE). In Experiment 1, we generated nulliparous (NP), primiparous (PP), and multiparous (MP) females (none, one, and two litters, respectively). Two weeks following the weaning of the first/second six-pup litters, the age-matched MP and PP and the non-pup-exposed NP animals were subjected to a 60-min restraint stress paradigm (enclosure in a Plexiglas restraint tube). The brains were removed and processed for c-fos immunoreactivity ( c-fos-IR) in CA3 region of the hippocampus (HI) and in basolateral amygdala (BLA). MP and PP females had very similar numbers of c-fos-IR neurons in both HI and BLA, whereas both were lower than NPs. In a second experiment, the same groups were generated, together with primigravid (PG; first pregnancy) and multigravid (MG; second pregnancy) females, tested in late pregnancy. The animals were exposed to a 30-min trial in an open field and were killed, and the brains were again examined for c-fos-IR. The parous and gravid animals displayed less reactivity to the stress of the open field (i.e., reductions in behavioral measures of anxiety) and significantly less c-fos expression in both CA3 and BLA. The gravid animals displayed significantly less c-fos expression in CA3 and BLA compared to parous females, although neither group differed as a result of a second RE. The data suggest that reproductive (viz., hormonal) and/or maternal (viz., pup exposure) experience may inure a female and her brain to stress, rendering her less susceptible to the behavioral—or other—disruptions that stress sensitivity can produce. Together, these data suggest that the experiences of motherhood (pregnancy, pup exposure, suckling stimulation, etc.) summate to produce reductions in anxiety and stress responsiveness that start before and last long after pup exposure and care. Such reductions may be adaptive in the face of demands placed upon the parous vs. the NP female.

Neuroendocrinology of Maternal Behavior in the Rabbit

Rabbit maternal behavior consists of building an underground nest of straw and body hair during late pregnancy and displaying, with circadian periodicity, a single 3-min nursing bout/day across lactation. Estrogen, androgen, progesterone, and prolactin regulate specific aspects of nest-building and promote the onset of maternal responsiveness. However, the maintenance of this behavior relies on stimuli from the litter: by preventing mother/young contact at parturition or during early lactation maternal responsiveness is altered or abolished. The brain areas controlling the expression of nest-building and nursing were investigated by implanting estradiol, locating the distribution of estrogen and prolactin receptors, quantifying the expression of immediate-early genes, and lesioning structures of the olfactory system. These studies revealed that: (a) estrogen receptor-α, alpha, present in the preoptic region, may mediate the stimulation of nest-building by estradiol; (b) prolactin binding sites, located mainly in periventricular structures, are more abundant in late pregnancy and early lactation; (c) the number of FOS-immunoreactive neurons increases in the lateral septum, but not in the mediobasal hypothalamus, following nursing; (d) the accessory olfactory bulb tonically inhibits the expression of maternal behavior because its removal promotes maternal responsiveness in virgins, which are otherwise unresponsive to daily pup exposure. In summary, rabbits rely on the same hormonal and extrahormonal factors that stimulate maternal behavior in other mammals, yet the way in which such factors promote elaborate nest-building and the unfailing display of circadian nursing is unique to rabbits and warrants future investigation.

Lack of Differential Sensitivity to Cholinesterase Inhibition in Fetuses and Neonates Compared to Dams Treated Perinatally with Chlorpyrifos

Pregnant Sprague-Dawley rats were exposed to chlorpyrifos (CPF; O,O-diethyl-O-[3,5,6-trichloro-2-pyridinyl] phosphorothioate) by gavage (in corn oil) from gestation day (GD) 6 to postnatal day (PND) 10. Dosages to the dams were 0 (control), 0.3 (low), 1.0 (middle) or 5.0 mg/kg/day (high). On GD 20 (4 h post gavage), the blood CPF concentration in fetuses was about one half the level found in their dams (high-dose fetuses 46 ng/g; high-dose dams 109 ng/g). CPF-oxon was detected only once; high-dose fetuses had a blood level of about 1 ng/g. Although no blood CPF could be detected (limit of quantitation 0.7 ng/g) in dams given 0.3 mg/kg/ day, these dams had significant inhibition of plasma and red blood cell (RBC) ChE. In contrast, fetuses of dams given 1 mg/kg/day had a blood CPF level of about 1.1 ng/g, but had no inhibition of ChE of any tissue. Thus, based on blood CPF levels, fetuses had less cholinesterase (ChE) inhibition than dams. Inhibition of ChE occurred at all dosage levels in dams, but only at the high-dose level in pups. At the high dosage, ChE inhibition was greater in dams than in pups, and the relative degree of inhibition was RBC approximately plasma > or = heart > brain (least inhibited). Milk CPF concentrations were up to 200 times those in blood, and pup exposure via milk from dams given 5 mg/kg/day was estimated to be 0.12 mg/kg/day. Therefore, the dosage to nursing pups was much reduced compared to the dams exposure. In spite of exposure via milk, the ChE levels of all tissues of high-dosage pups rapidly returned to near control levels by PND 5.

Expression of μ-opioid receptor mRNA in the medial preoptic area of juvenile rats

Previous studies indicate that the latency to initiate parental behavior in both male and female rats increases with age; weanling (21 days old) rats display parental behavior 0–2 days after exposure to newborn pups, while older juveniles (30 days old) require 5–6 days of pup exposure before they express the behavior. Furthermore, activation of μ-opioid receptors inhibits parental behavior in juvenile and adult rats. We hypothesized that the age-related increase in behavioral latency could be modulated by the induction of μ-receptor expression in the medial preoptic area (MPOA), a region in which μ-receptors regulate parental behavior. In situ hybridization histochemistry was used to measure μ-receptor mRNA expression in the MPOA of male and female Sprague–Dawley rats that were 21, 24, 27, 30, or 33 days old. Using autoradiographic film analysis, we observed that neurons within part of the MPOA expressed very dense μ-receptor mRNA. Comparison of mRNA distribution with histological boundaries indicated that neurons within the medial preoptic nucleus (MPN), excluding the central part, exhibited the highest density of μ-receptor mRNA within the MPOA. High densities of μ-receptor mRNA extended dorsolaterally and caudally from the MPN toward the bed nucleus of the stria terminalis. MPN μ-receptor mRNA expression was not altered with age and no sex difference was observed. The dense presence of μ-receptor mRNA in the MPN suggests that ample substrate exists on which μ-receptor ligands could modulate the latency to begin parental behavior in juvenile rats, but such behavioral expression apparently is not mediated by a change in μ-receptor mRNA production.

Using c-Fos immunocytochemistry to identify forebrain regions that may inhibit maternal behavior in rats.

Evidence indicates there is a neural system that inhibits maternal behavior in virgin rats. It has been suggested that pregnancy hormones promote the onset of maternal behavior by reducing the behavioral influence of this system. The authors used c-Fos immunocytochemistry to identify brain regions more activated by pup exposure in nonmaternal rats than in maternal rats. Previous experiments indicated that some of these regions, such as the posterodorsal medial amygdala and several medial hypothalamic sites, inhibit maternal behavior. For others, such as the ventral lateral septum, dorsal premammillary nucleus, and principal bed nucleus of the stria terminalis, this is the first indication that they could also inhibit maternal responding. These regions have previously been implicated in promoting defensive behaviors, consistent with the finding that nonmaternal rats actively avoid pups. These findings suggest the existence of a neural circuit through which pup exposure could promote defensive responses in virgin rats, and how pregnancy hormones could reduce such activity to stimulate maternal behavior.

Using c-Fos immunocytochemistry to identify forebrain regions that may inhibit maternal behavior in rats.

Evidence indicates there is a neural system that inhibits maternal behavior in virgin rats. It has been suggested that pregnancy hormones promote the onset of maternal behavior by reducing the behavioral influence of this system. The authors used c-Fos immunocytochemistry to identify brain regions more activated by pup exposure in nonmaternal rats than in maternal rats. Previous experiments indicated that some of these regions, such as the posterodorsal medial amygdala and several medial hypothalamic sites, inhibit maternal behavior. For others, such as the ventral lateral septum, dorsal premammillary nucleus, and principal bed nucleus of the stria terminalis, this is the first indication that they could also inhibit maternal responding. These regions have previously been implicated in promoting defensive behaviors, consistent with the finding that nonmaternal rats actively avoid pups. These findings suggest the existence of a neural circuit through which pup exposure could promote defensive responses in virgin rats, and how pregnancy hormones could reduce such activity to stimulate maternal behavior.