Joseph S. Lonstein

Sensory, hormonal, and neural control of maternal aggression in laboratory rodents.

Parental animals of many rodent species display fierce and persistent aggression toward unfamiliar conspecifics that appears to protect their often altricial and defenseless young. We herein review studies of the sensory, hormonal, neuroanatomical, and neurochemical mechanisms underlying maternal aggression in laboratory rodents. The relationship between maternal aggression and fearfulness or anxiety is also discussed.

Sex differences in the parental behavior of rodents

The reproductive strategy of many mammalian species that give birth to altricial young involves intense and prolonged care of their offspring. In most cases, the mother provides all nurturance, but in some cases fathers, older siblings, or unrelated conspecifics participate in parental care. The display of these behaviors by animals other than mothers is affected by numerous factors, including their sex. We herein review the literature on similarities and/or differences between male and female laboratory rodents (rats, mice, voles, gerbils, and hamsters) in their parental responsiveness and discuss how the parental behavior of males and females is influenced by hormones, developmental processes, and prior social experiences. Understanding the mechanisms that generate sex differences in the parental responsiveness of rodents may indicate how similar sex differences in parental care are generated in other mammals.

Regulation of anxiety during the postpartum period

Healthy mother-infant interactions are critical for the physical, cognitive, and psychological development of offspring. Such interactions rely on numerous factors, including a positive maternal emotional state. However, many postpartum women experience emotional dysregulation, often involving elevated anxiety. Neuroendocrine factors contributing to the onset of postpartum anxiety symptoms are mostly unknown, but irregularities in hypothalamic-pituitary-adrenal axis function, reduced prolactin and oxytocin signaling, or parturitional withdrawal of ovarian, placental and neural steroids could contribute to anxiety in susceptible women. Although the causes of initial onset are unclear, postpartum anxiety can be mitigated by recent contact with infants. Numerous neurochemical systems, including oxytocin, prolactin, GABA, and norepinephrine mediate this anxiolytic effect of infant contact. Insight into the etiology of postpartum anxiety disorders, and how contact with infants helps counter existing anxiety dysregulation, will surely facilitate the diagnosis and treatment of postpartum women at risk for, or experiencing, an anxiety disorder.

Sex differences in the parental behaviour of adult virgin prairie voles: Independence from gonadal hormones and vasopressin

Sexually and parentally experienced prairie voles display robust biparental care of pups that is similar between the sexes. Little is known, however, about possible sex differences in the parental behaviours of sexually inexperienced prairie voles. Parental behaviour of adult virgin male and female prairie voles was examined in sham‐operated and gonadectomized subjects treated with vehicle or oestradiol. Since arginine‐vasopressin (AVP) has been suggested to stimulate parental behaviour in sexually inexperienced males, neural AVP immunoreactivity (AVP IR) was quantified. Most sham‐operated and castrated males displayed high levels of parental behaviour (9/9 controls, 6/9 castrates) during a 15‐min exposure to pups 4 weeks after surgery, and few behavioural differences were seen between groups. Conversely, almost all gonadally intact (8/9) and gonadectomized (8/9) females attacked pups. Implantation of a 0.1‐mg pellet of oestradiol immediately after gonadectomy had little effect on males (9/9 parental), whereas most (5/9) oestradiol‐treated females acted maternally. AVP‐immunoreactive (AVP‐ir) fibre density in the lateral septum (LS) and lateral habenula (LHb), expressed by the number of pixels that covered AVP‐ir fibres during computerized optical density analysis, was greater in males than females, was non‐significantly reduced in castrated males, and doubled in the LS of oestradiol‐treated females. In a second experiment, males tested 8 weeks after similar manipulations remained highly parental though castrated males had almost no AVP‐ir fibres in the LS and LHb. Levels of AVP IR in males treated with oestradiol were similar to those observed in intact males. A dramatic sex difference therefore exists in the parental behaviour of adult sexually naive prairie voles which cannot be explained by sex differences in gonadal hormones. Because both castrated and intact males were highly parental, even though castrates had virtually no AVP‐ir in the LS or LHb, AVP does not appear to be crucial for their responsiveness toward pups.

Comparison of the parental behavior of pair-bonded female and male prairie voles (Microtus ochrogaster)

The behavior of primiparous lactating prairie voles (Microtus ochrogaster) and their mates individually interacting with pups was continuously assessed for 45 min after a 2-h parent-litter separation on days 3-4 and 10-11 postpartum. Both sexes were highly parental after reunion with the young, and their general pattern of behavior consisted of bouts of quiescence interspersed with bursts of heightened activity. Lactating females spent more time than males in contact with pups, and more time being quiescent, most often in the kyphotic (upright crouched) nursing posture. Even in the absence of nipples upon which the pups could suckle, males also displayed kyphosis, although for shorter durations than females. Males spent more time, however, huddled over the litter in a hunched position than their mates. In accordance with their decreased quiescence, male voles licked and carried pups more and were more exploratory than females. Compared with the first week postpartum, bouts of kyphosis were shorter during the second week postpartum for both sexes, while laying prone on the pups increased. Males spent less time licking and more time carrying older pups than younger ones, and were more exploratory during the second week postpartum. Sex differences in the parental behavior of prairie voles may reflect differences in the somatosensory stimulation that females and males receive from pups. Furthermore, the display of kyphosis by male voles indicates that the sensorimotor organization of this posture in voles differs from that of lactating rats, which require suckling stimulation for its regulation.

Neural mediation of nursing and related maternal behaviors.

Nursing is the behavioral concomitant of lactation and the most generalizable maternal behavior across mammals. In lactating rats nursing often occurs in the kyphotic (upright crouched) posture; like the neuroendocrine determinants of milk synthesis and release, kyphosis requires suckling by the young. The dams active pronurturant behaviors, such as retrieval and licking of pups, requires perioral somatosensory stimulation, which is often a precursor of kyphosis as well, and is inhibited by suckling. The sequential nature of maternal behaviors and the dissociations in their somatosensory regulation are critical to understanding their neural mediation, as exemplified by our recent work in lactating rats. We found that the caudal lateral and ventrolateral midbrain periaqueductal gray (cPAGl,vl) is a sensorimotor integration site for the kyphotic nursing posture. Destruction of the cPAGl,vl, or increased activity of the inhibitory neurotransmitter GABA within it, severely reduced kyphosis, increased nursing in more atypical postures, and had little or no effect on pronurturance. Various forebrain sites are known to mediate retrieval and licking of pups. Inhibition of dopaminergic activity in the nucleus accumbens of dams via microinfusions of a mixed D1/D2 dopamine receptor antagonist, cis-flupenthixol (FLU), dose-dependently reduced these active behaviors, while increasing nursing duration. Retrieval was inhibited, however, only by infusions of FLU that included the nucleus accumbens shell, which is reciprocally connected with other sites implicated in retrieval of pups. Thus, maternal behavior is not a unitary process but rather a complex category consisting of sequential behavioral components that have their own sensory and neural determinants.

Maternal behavior stimulates c-fos activity within estrogen receptor alpha-containing neurons in lactating rats

Estradiol and other hormones are thought to be critical for the onset, but not maintenance, of maternal behavior in rats. Maternal behavior is instead maintained postpartum by tactile stimulation that dams receive during interactions with pups, and many neural sites implicated in the control of maternal behavior show elevated c-fos activity in response to this stimulation. Many of these sites also contain neurons that express the alpha subtype of the estrogen receptor (ERα). Because of possible interactions between tactile stimulation from pups, c-fos, and ERα in the lactating rat brain, we determined if populations of cells that show increased c-fos activity after maternal behavior in lactating rats also contain ERα. Dams were separated from their pups for 48 h beginning on day 5 postpartum. On day 7 postpartum, experimental dams were reunited with pups and mother-litter interactions were observed for 60 min. Control dams received no pup stimulation. Subjects were sacrificed 60 min later and brain sections were double immunolabeled for the Fos and ERα proteins. As expected, the number of ERα-immunoreactive (ERα-ir) neurons did not differ between the two groups in the eight areas analyzed (lateral region of the lateral septum, posterodorsal medial amygdala, dorsal and ventral medial preoptic area, dorsal and ventral bed nucleus of the stria terminalis, lateral habenula, and ventrolateral caudal periaqueductal gray). Consistent with previous reports, maternal dams had 2- to 7-fold more Fos-immunoreactive (Fos-ir) neurons in these sites compared with nonstimulated controls. Maternal dams had significantly more Fos-ir neurons that also contained ERα-ir in all sites, with the greatest increases in the ventral medial preoptic area, lateral habenula, and ventral bed nucleus of the stria terminalis. Between ∼25 and 45% of the Fos-ir cells in the sites examined also expressed ERα. Thus, a substantial number of neurons that are genomically activated during maternal behavior contain ERα, raising the possibility that the postpartum display of maternal behavior can be influenced by ERα activity.

MATERNAL BEHAVIOUR IN LACTATING RATS STIMULATES c-fos IN GLUTAMATE DECARBOXYLASE-SYNTHESIZING NEURONS OF THE MEDIAL PREOPTIC AREA, VENTRAL BED NUCLEUS OF THE STRIA TERMINALIS, AND VENTROCAUDAL PERIAQUEDUCTAL GRAY

Increased activity of the immediate-early gene c-fos can be observed in many areas of the lactating rat brain after dams physically interact with pups and display maternal behaviour. These sites include the medial preoptic area, ventral bed nucleus of the stria terminalis, and the ventrolateral caudal periaqueductal gray, each of which is critical for the normal performance of particular maternal behaviours. The phenotype of cells in these areas that show increased c-fos activity after maternal behaviour, however, is unknown. Via double-label immunocytochemistry, we determined if the population of cells in these sites that express c-fos after maternal behaviour in lactating rats overlaps with the population that expresses the 67,000 mol. wt isoform of glutamate decarboxlyase, the synthesizing enzyme for the inhibitory neurotransmitter GABA. Lactating rats were separated from pups beginning on day 5 postpartum, and 48h later half were allowed to interact with a litter of pups for 60min whereas the other half were not. Dams re-exposed to pups were highly maternal, retrieving and licking them as well as displaying prolonged nursing behaviour that included milk letdown. Both groups of dams had a similar number of 67,000 mol. wt glutamate decarboxylase-immunoreactive cells in each site, although the number of 67,000 mol. wt glutamate decarboxylase-immunoreactive cells per microscopic field was significantly greater in the caudal ventrolateral periaqueductal gray than in the ventral bed nucleus of the stria terminalis, which in turn was greater than the medial preoptic area. In pup-stimulated dams, two to fourfold more Fos-immunoreactive cells were found in these three sites compared with non-stimulated controls. Labeling for Fos immunoreactivity and 67,000 mol. wt glutamate decarboxylase immunoreactivity was heterogeneous within each site. In the medial preoptic area, more Fos-immunoreactive and 67,000 mol. wt glutamate decarboxylase-immunoreactive cells (either single or dual-labeled) were found dorsally than ventrally. In the ventral bed nucleus of the stria terminalis, more Fos-immunoreactive and 67,000 mol. wt glutamate decarboxylase-immunoreactive cells were found medially than laterally. Within the caudal ventrolateral periaqueductal gray, 67,000 mol. wt glutamate decarboxylase-immunoreactive labeling was greatest ventromedially, while high numbers of Fos-immunoreactive nuclei were found both ventromedially and ventrolaterally. In pup-stimulated dams, more than half (53% in the medial preoptic area, 59% in the ventral bed nucleus of the stria terminalis, and 61% in the caudal ventrolateral periaqueductal gray) of the total population of Fos-immunoreactive cells also expressed 67,000 mol. wt glutamate decarboxylase. These results suggest that many of the neurons in these sites that show elevated c-fos activity after maternal behaviour are either local inhibitory interneurons or provide inhibitory input to other neural sites. These inhibitory mechanisms may be critical for the display of postpartum nurturance, possibly facilitating maternal behaviour by removing tonic inhibition on sites necessary for maternal responding or by restricting activity in neural sites that inhibit it.

Dopamine D1 and D2 Receptor Antagonism in the Preoptic Area Produces Different Effects on Maternal Behavior in Lactating Rats

The preoptic area (POA) is critical for maternal behavior in rats but little is known about what neurotransmitters released here influence maternal responding. POA infusion of 10 microg (but not 2 microg) of the dopamine D1 receptor antagonist SCH-23390 greatly impaired retrieval and licking of pups but not other maternal or nonmaternal behaviors in lactating rats. In contrast, POA infusion of 10 microg (but not 2 microg) of the D2 receptor antagonist raclopride facilitated nursing but did not affect oral maternal behaviors. SCH-23390 in the medial hypothalamus tended to impair licking but not retrieval. Raclopride in the medial hypothalamus had no effects. Therefore, D1 and D2 receptor activity, particularly in the POA, is important for regulating different maternal behaviors.

Effects of neonatal RU486 on adult sexual, parental, and fearful behaviors in rats.

Exposure to gonadal hormones during perinatal life influences later behavior. The finding that sex differences exist in progestin receptor expression in the perinatal rat brain suggests differential sensitivity of male and female brains to progesterone (C. K. Wagner, A. N. Nakayama, & G. J. De Vries, 1998). Because these sex differences are in neural sites that influence sexually differentiated sexual, parental, and fearful behaviors in adults, this study examined the effects of administering the progestin receptor antagonist RU486 for the first 10 days after birth on these behaviors in adulthood. Neonatal RU486 significantly reduced sexual behavior in males but did not impair reproduction in females. Neonatal RU486 did not affect parental responses of virgin rats exposed to pups (sensitization) but reduced fear in the elevated plus-maze in both sexes. Treatment of pups with RU486 affected neither mother-litter interactions nor plasma testosterone levels in males during or after treatment. These results suggest that neonatal exposure to progesterone, in addition to androgens and estrogens, influences behavioral development in rats.