Sex steroid hormone determination of the maternal brain: effects beyond reproduction.

Herein we discuss the effects of hormones on reproduction, but with a focus on the ripples that emanate from the main effects. That is, the role of hormones in reproductive events is both well-known and well accepted; less studied and understood are effects that appear to be ancillary to the primary objectives of the hormonal effects, which support, complement and extend their primary effects. We present evidence for how the hormonal stimulation of pregnancy constructs the maternal brain; makes it more efficient; enhances cognition; regulates stress responsiveness; modifies sensory systems (we discuss mainly olfaction); neurogenesis; and learning. Thus, steroid and other hormones and neuropeptides restructure the nervous system, particularly of females, to produce and regulate maternal behavior as well as behaviors and physiological systems that contribute to and support what is arguably the primary function of the hormones: survival and effective nurturance of the female's metabolic and genetic investment.

Fatherhood alters behavioural and neural responsiveness in a spatial task.

The hormones and experiences of pregnancy, parturition and lactation have been shown to dramatically remodel the female rat's hippocampus, potentially enhancing behaviours critical for meeting the increased demands of motherhood. Previous work in our laboratory has also suggested that pup exposure, apart from pregnancy and lactation, constitutes an important influence on ancillary maternal behaviour (e.g. foraging behaviour). In the present study, we press the parental model further by examining the effect of pup exposure on the hippocampus of males from a biparental mouse species, the California mice (Peromyscus californicus). Males were either Fathers (i.e. first-time fathers housed with a female from mating until 7 days after parturition), pup-exposed virgins (PEV; i.e. sexually naïve males briefly exposed to pups daily for 7 days) or Virgins (i.e. never exposed to females or pups). A dry-land maze (DLM), as used for assessing spatial learning, was employed to determine the foraging abilities of the males. The results indicated that, on the most challenging day of testing (i.e. acquisition day), California mouse Fathers demonstrated superior memory for the task compared to PEVs and Virgins. In addition to the behavioural data, significantly more fos-immunoreactivity was observed in the CA1, CA3 and dentate gyrus regions of the hippocampi of Fathers than PEVs or Virgins in response to the probe trial. Additionally, a trend for altered performance on the DLM was observed in the PEVs on the last day of testing, which was accompanied by the highest levels of nestin-immunoreactivity, an indicant of neuroplasticity, of the three groups. In summary, these data suggest that, in accordance with previous observations of maternal rats, the paternal brain is similarly influenced by parental experience, as demonstrated by accompanying modifications to relevant neurobiological and behavioural responses.

Why the maternal brain?

In the rat, the change from a virgin/nulliparous female to the maternal animal takes place at many levels. A subtle developmental wave washes over the female nervous system and transforms her from largely self-centred to offspring-directed, from personal care and protection to care of genetically-related offspring, from indifference to ardour. Such change is preceded by substantial and apparently permanent neural alterations, the depth of which results in the maternal brain, and is the basis of the present review. The neuroplasticity of pregnancy, inherent to the female brain and, we believe, representative of the full expression of the female nervous system's capacity, is a result of significant hormonal and other neurochemical actions. It results in the striking brain changes that are associated with, and necessary for, successful reproduction. We discuss some of these changes and their ramifications. Collectively, they represent the culmination of mammalian evolution and have led to the development of the social brain characteristic of higher orders of mammal, including the human. We also examine different facets of the maternal brain, beginning with a review of the genes involved in maternal behaviour, and in the subsequent 'expression' of the maternal brain. We next discuss olfaction and the manner in which this major sense draws from the rich sensory milieu of the mother to regulate and support maternal behaviour. Last, we discuss the 'whys' of maternal behaviour, a theoretical foray into the reasons for such substantial maternal brain alterations. We focus on the male's potential role as the raison d'etre for the manifest alterations in his mate's brain. In the end, it is clear that the female brain undergoes a significant reorganisation en route to motherhood, the results of which are deep and enduring.

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.

Alterations of medial preoptic area neurons following pregnancy and pregnancy-like steroidal treatment in the rat.

There is a marked increase in the maternal behavior displayed by a female rat following pregnancy-due primarily to exposure to the gonadal hormones progesterone and estradiol (P and E(2), respectively). We examined Golgi-Cox silver-stained, Vibratome-sectioned neurons visualized and traced using computerized microscopy and image analysis. In Part One, we examined the hormonal-neural concomitants in the medial preoptic area (mPOA), an area of the brain that regulates maternal behavior, by comparing cell body size (area in microm(2); also referred to as soma and perikaryon) in the mPOA and cortex of five groups (n = 4-6/group) of ovariectomized (OVX-minus), diestrous, sequential P and E(2)-treated (P+E(2)), late-pregnant, and lactating rats; for Part Two, we examined a subset of mPOA neurons, which were traced in their entirety, from these same subjects. In Part One, whereas there was no difference between OVX-minus and diestrous females, both had smaller somal areas compared to OVX+P+E(2)-treated and late-pregnant females. The area of the soma returned to diestrous/OVX-minus levels in the lactating females. We found no change among the five groups in area of cell body in cortical neurons, which generally lack steroid receptors. In Part Two, which included a more detailed morphometric analysis of mPOA neurons, we examined several additional measures of dendritic structure, including number of proximal dendritic branches (the largest proximal dendrite was defined as the one with the largest diameter leaving the soma); cumulative length of the largest proximal dendrite; area of the cell body; number of basal dendrites; cumulative basal dendritic length; number of basal dendritic branches; and branch-point (distance from cell body to first branch of largest proximal dendrite). Again, we found similar effects on cell body size as in Part One, together with effects on number of basal dendritic branches and cumulative basal dendritic length in pregnant and P+E(2)-treated groups compared to OVX, diestrous, and lactating. An increase in somal area denotes increased cellular activity, and stimulatory effects on additional neuronal variables represents modifications in information processing capacity. Pregnancy and its attendant hormonal exposure, therefore, may stimulate neurons in the mPOA, which then contribute (in an as yet undetermined manner) to the display of maternal behavior. During the postpartum lactational period, when cues from pups primarily maintain maternal attention, the neuronal soma appears to return to a pre-pregnancy, non-hormonally dependent state, whereas other aspects of the dendrite remain altered. Collectively, these data demonstrate a striking plasticity in the brains of females that may be reflected in modifications in behavior.

Activity-stress increases density of GFAP-immunoreactive astrocytes in the rat hippocampus.

Although past research has indicated that stress and the accompanying increase in glucocorticoids compromises hippocampal neurons, little is known about the effect of stress on hippocampal glial cells. In the current study, male rats were exposed to activity-stress (A-S) for six days; this comprised housing with an activity wheel and restricted access (1h/day) to food. Physiological data (e.g., relative adrenal and thymus weights, gastric ulceration) suggested that the A-S rats experienced more stress than pair-fed (no wheel) and control (fed ad libitum, no wheel) rats. Whereas stress did not influence the quantitative morphology of glial fibrillary acidic protein (GFAP)-immunoreactive cells, a semi-quantitative analysis revealed that the A-S rats had significantly more (30%) GFAP-immunoreactive cells in the hippocampal CA3 region than the control rats. Based on the present findings, it appears that the hippocampal astrocytic response to chronic stress may be similar to the response found in endangered, or challenged hippocampal environments, such as in ischemia.

Activity-stress induces atrophy of apical dendrites of hippocampal pyramidal neurons in male rats.

Recently, researchers have demonstrated the damaging effect of restraint-stress on hippocampal neurons. The purpose of the present study was to determine if a more chronic stressor, i.e., activity-stress (A-S), would also result in hippocampal dendritic atrophy. When activity-stress (n = 6) rats showed evidence of the criteria "stress symptoms" (after an average of 6 days), they were sacrificed and their brains were quickly removed, blocked, and placed in Golgi-Cox solution. Food-yoked control animals (n = 6) were sacrificed on the following day. Serial coronal sections (150 um) of the rostral hippocampus were cut so that the CA3 and CAI areas could be analyzed. Stressed short-shaft neurons were significantly shorter and had fewer branch points in CA1 and CA3 neurons than the control neurons. A similar nonsignificant trend was observed in long-shaft neurons. These data suggest that a short period of chronic stress (6 days as opposed to 21 days in prior studies) induces neuronal atrophy in the hippocampus.

Opiate disruption of maternal behavior: morphine reduces, and naloxone restores, c-fos activity in the medial preoptic area of lactating rats.

Morphine significantly impairs maternal behavior; naloxone, an opiate antagonist, restores it. Maternal behavior is associated with c-fos expression, an immediate early gene product, in the medial preoptic area (mPOA) of females. In two experiments, the effects of morphine-alone and morphine plus naloxone on the expression of c-fos were examined. On postpartum day 5, females were injected with morphine or saline (experiment 1), and morphine + naloxone or morphine + saline (experiment 2) and placed back in the home-cage, separated from their pups by a wire-mesh partition. A separate group in experiment 1 was injected but not exposed to pups. Processing for c-fos immunohistochemistry commenced, and c-fos positive cells in a proscribed portion of mPOA were counted. Morphine-treated females had fewer c-fos cells in mPOA compared to saline-treated females, and the presence of pups accounted for a significant increase in c-fos-expressing neurons, whereas in females not exposed to pups, morphine treatment did not significantly reduce baseline c-fos expression (experiment 1). Furthermore, naloxone mitigated the effect as morphine + naloxone-treated females expressed more c-fos cells compared to morphine + saline females (experiment 2). Morphine-treated females, therefore, may exhibit reductions in maternal behavior because of relative opiate-induced inactivation of areas of the brain devoted to the regulation of maternal behavior.

Prenatal stress alters the size of the rostral anterior commissure in rats.

In rodents and other mammals, prenatal stress disrupts both sexual differentiation and sexual behavior. The present study examined the area of the anterior division of the anterior commissure (the Aca) in coronal, thionin-stained sections of prenatally stressed (P-S), and control male and female rats. Pregnant rats were exposed to thrice-daily heat, light, and restraint stress or left undisturbed during days 15-22 of pregnancy. Adult P-S and control males and females were killed, perfused, and their brains removed. Serial coronal sections (total of approximately 200 microm) through the rostral portion of the Aca (the rAca) were taken and stained with thionin. The sections were examined and traced under x25 using computerized microscopy to obtain the area in mm2. The data revealed that control females had a larger rAca compared to control males, and that P-S males had a larger rAca compared to control males; further, control males and P-S females were not significantly different, nor were control females and P-S males. These results suggest that, in rats, the Ac may be sexually dimorphic (in a direction similar to that described in humans) and that prenatal stress an event that modifies sex-typical behavior, physiology, and neuroanatomy reverses that sex difference.

Preoptic area infusions of morphine disrupt--and naloxone restores--parental-like behavior in juvenile rats.

As in the adult lactating female, opioids disrupt (and naloxone restores), parental behavior in juvenile rats (approximately 25 days of age). Because the preoptic area regulates the display of parental behavior in lactating females, we examined its parental behavior role in the juvenile rat. At 21 days of age, juvenile rats were implanted with bilateral cannulae aimed at the preoptic area using a modified Kopf stereotaxic and extrapolating from a developing-rat brain atlas [58], and divided into two groups: Initiation and maintenance. On day 25, the initiation group received bilateral infusions of either morphine (0.50 microgram), saline (0.25 microliter), or morphine plus naloxone (0.25 microgram). Thirty minutes later, they were exposed to three 1-6-day-old pups; the maintenance group was exposed to pups until they displayed 2 consecutive days of parental behavior, then infused. Morphine disrupted parental behavior in both the initiation and Maintenance groups, and naloxone restored the behavior to control/ saline levels. Parental behavior in the juvenile animal of both sexes, therefore, is under opioid regulation that parallels the adult female.

Prenatal stress attenuates ulceration in the activity stress paradigm.

In the present study, 28 pregnant rats were subjected to either light-restraint stress or no manipulation for days 14-21 of the gestational period. At approx. 50 days of age, both male (n = 16) and female (n = 16) prenatally stressed (PS) and control offspring were subjected to the activity stress (AS) paradigm. During this subsequent stress experience, PS rats developed less ulceration than control rats. PS rats also displayed about half the activity of the control animals during the habituation phase of the AS paradigm, prior to the induction of stress. Given this decrease in baseline activity in PS animals, implications of using activity as a measure of emotionality in PS animals are discussed. Several sex differences were also observed; females differed from males in that they 1) exhibited higher activity levels in both the habituation and experimental phases of the AS procedure, 2) developed heavier relative adrenal weights, and 3) reached criteria for sacrifice in fewer days.

Intracerebroventricular infusions of morphine, and blockade with naloxone, modify the olfactory preferences for pup odors in lactating rats.

Systemic morphine exposure disrupts both maternal behavior (MB) and postpartum aggression, possibly through alterations of olfactory preferences [29]. In the current studies, adult female rats were timed mated and implanted with a unilateral cannula in the lateral ventricle. On day 5 or 6 of lactation, the females were infused with either morphine (2.0 micrograms) or saline (5 microliters) (Experiment 1); or, they were infused with 2.0 micrograms morphine and saline or morphine plus 0.5 micrograms naloxone (Experiment 2). One hour later, they were exposed to bedding soiled by pups, or to clean bedding. The amount of time spent investigating the two bedding types was compared. Morphine produced an aversion to the odor of pups, relative to the saline condition, wherein the females expressed a preference for the odor of pups. Naloxone reversed the effect of the morphine, restoring the preference for pup odors in the females. Thus, morphine disruption of MB may be due to central action on olfactory sensory mechanisms.

Paucity of c-fos expression in the medial preoptic area of prenatally stressed male rats following exposure to sexually receptive females.

Normal male rats display high levels of sexual activity when paired with sexually receptive females, a behavior regulated, in large part, by the medial preoptic area (MPOA). It has been documented that onset of c-fos proteins in the MPOA accompanies sexual behavior. Because prenatal stress (PS) demasculinizes sexual behavior in male rats, the present study examined whether such effects might be accompanied by a decrease in the neuronal activation in MPOA associated with sexual behavior. Adult prenatally stressed (P-S) and unstressed control males were paired with sexually receptive females, and sexual behavior allowed to commence. After a single mount, the pair was separated by a partition for the remainder of an hour, at the end of which the male was killed and the brain processed for c-fos immunocytochemistry; quantification was performed by means of computerized image analysis. P-S males expressed significantly less c-fos activity in the region of the MPOA, compared to nonstressed control males. Thus, the sexual deficits associated with P-S males may be due to a relative dearth of activity in the MPOA at the neuronal level. Coupled to earlier work showing decreased luteinizing hormone secretion in P-S males under similar conditions, the present data suggest a constellation of factors that contribute to PS-induced deficits in sexual behavior.

Cocaine alters the onset and maintenance of maternal behavior in lactating rats.

Though much attention has been devoted to the behavioral and physiological consequences of cocaine abuse in offspring, little is known regarding the effects on the maternal behavior of the cocaine-exposed dam. We examined whether cocaine affects the initiation (late pregnancy) and/or maintenance (postpartum [PP]) phases of full maternal behavior (FMB; retrieving, grouping, and crouching over six pups) in Sprague-Dawley female rats. In Experiment 1, cocaine (5.0 or 10.0 mg/kg) or saline was administered on PP day 5 or 6 and FMB scored. Both dosages significantly disrupted FMB, particularly crouching, though 10.0 mg/kg had a greater effect on FMB. Experiment 2 (using 10.0 mg/kg cocaine) examined specific elements of the disruption and found significant reductions in proportion of females engaging in FMB, as well as increases in the latencies to contact, retrieve, lick, group, and crouch over pups. In Experiment 3 osmotic pumps containing 20 mg cocaine/kg/day or saline were implanted SC in day 14 pregnant rats. FMB testing was performed on days 1-2 postpartum together with a T-maze pup-retrieval test on postpartum days 3-5. Cocaine disrupted FMB in the homecage, in general, rendering the females less attentive to young, but was without effect in the T-maze tests. Cocaine--perhaps owing to its purported dopaminergic activity--may operate through motivational mechanisms to disrupt FMB in the postpartum maintenance phase; and through effects on late pregnancy levels of prolactin (a hormone which stimulates FMB), to disrupt maternal behavior during the initiation phase.

Developmental psychobiological influences on rodent parental behavior.

The present paper focuses on the development, broadly defined, of parental responsiveness. Both sexes can respond to young with parental behavior, in the absence of elaborate experimental manipulations; yet, through the combined natural forces of ontogeny and maturation, organizational influences of hormones, and adult modifications of neurochemical sensitivity, these animals can be rendered more or less sensitive to young. We will explore the ways in which such developmental alterations determine whether or not an animal is responsive to neonates.

Sex differences and gonadal hormones influence susceptibility to the activity-stress paradigm.

The present study investigated the effect of gonadal hormones on activity, food consumption, severity of ulceration, weight loss, and survival duration in male and female rats exposed to the activity-stress (A-S) paradigm. Animals received either sham surgery or gonadectomy and were housed in activity wheels or standard suspended cages. Results indicated that gonadectomized animals were more likely to engage in high running levels in the A-S paradigm; in addition, castrated males developed significantly more ulceration than the other groups. Neither sex nor gonadectomy affected the survival duration of A-S animals.

Opioid regulation of parental behavior in juvenile rats.

When exposed to young rats for a period of days, juvenile rats will respond with full parental behavior (FPB: retrieval and grouping of, and crouching over, pups). Because the parental behavior of juveniles is so robust, and because opiates have been shown to be involved in the regulation of parental behavior in adult animals, we examined morphine's ability to disrupt the display of parental behavior in the juvenile animal. In Experiment 1, 25-day-old males and females were administered one of three injection regimens of morphine (MOR), saline (SAL), or naloxone (NAL)--[MOR (5.0 mg/kg) + SAL; MOR + NAL (0.5 mg/kg); or SAL + SAL]--and 1 h later were exposed to three 1- to 6-day-old neonates. Behavior was scored over a 60-min period and animals were considered parental if they responded with FPB for 2 consecutive testing days. Whereas the SAL + SAL and MOR + NAL were not different, MOR + SAL virtually failed to respond to young over a 10-day period. Experiment 2 exposed juveniles to young for a period of days until they displayed 2 consecutive days of FPB. Next, separate groups of juveniles were treated with the same regimen as above. Again, SAL + SAL and MOR + NAL responded rapidly to young, whereas MOR + SAL did not. These data suggest that the display of parental behavior, both its onset (Experiment 1) and maintenance (Experiment 2), appears to be regulated by opiates.