Bicuculline infusion into the accessory olfactory bulb facilitates the induction of maternal behavior in rats

The GABAA antagonist bicuculline, intracranially infused into the accessory olfactory bulb (AOB), facilitated the expression of maternal behavior (MB) in virgin Wistar female rats. Behavioral effects were observed 24 hours after infusion and were injection dependent. Pheromonal stimuli, generated by the pups, are thought to exert an inhibitory effect on vomeronasal nuclei involved in MB in virgin rats. The present study investigated the possibility that a decrement in AOB output, resulting from long-term compensatory synaptic changes to chronic bicuculline infusion, would facilitate the expression of MB. The implications of our findings for the mechanisms involved in the induction of MB and the maternal experience effect are discussed.     

Cycloheximide blocks the retention of maternal experience in postpartum rats

Two studies were done to determine the effects of cycloheximide (CYX), a protein synthesis inhibitor, on maternal experience effects in rats. In the first study eight groups received a 2-h maternal experience 36 h after cesarean (c)-section and two groups received no post c-section experience. Among the experienced groups, two received icv injections of CYX or saline (SAL) 30 min before the maternal experience, two received CYX or SAL 10 min after the experience, and two received the injections 24 h after the experience. One inexperienced group received CYX and the other received SAL 36 h after c-section. Tests for maternal behavior occurred 10 days after c-section. CYX was not able to block or disrupt the "acquisition" or expression of ongoing maternal behavior during the 2-h experience phase. However, CYX was able to block the long-term "retention" of a 2-h maternal experience if the drug was present during or immediately after the experience, prior to "consolidation." The second study investigated the effects of CYX administered immediately after the maternal experience on the expression and retention of maternal behavior 4 and 6 days after c-section, to determine whether the hormonally mediated short-onset latencies of the 4-day group would be blocked by CYX. Eight groups of animals were tested for maternal behavior. Four were tested 4 days after c-section and four were tested 6 days after c-section. Within each of these groups two were experienced and two inexperienced; within each experience condition one group received CYX and one received SAL. Day 4 groups exhibited shorter onset latencies than Day 6 groups. There was also a CYX-SAL difference in maternal onset latencies among experienced Day 6 groups but not among Day 4 groups. These data indicate that the blocking effects of CYX can be seen only when hormonal priming of maternal behavior is no longer in evidence.     

The role of the accessory and main olfactory systems in maternal behavior of the primiparous rat

The objective of this study was to analyze the respective roles of the main and accessory olfactory systems in the development of maternal behavior in the primiparous Wistar rat. Females underwent one of the following treatments: vomeronasal nerve section (VN), irrigation of nasal cavities with 5% ZnSO4 solution (ZN), surgical control, saline irrigation control, and normal control. Surgical or first irrigation were done before mating occurred. Irrigations were done every 7 days thereafter. The dams and their litters were observed from the day the litter was born (Day 1) through Day 16. Pup weights and temperatures were recorded daily. Home cages were checked daily for changes in nest location and number of times pups were found out of the nest. Retrieving tests were conducted on Days 4, 7, 10, and 13. The following behavioral items were observed: number of pup retrievals , number of times mother nosed or licked pups, percentage of litter returned to nest by end of test, dam self-grooming, dam climbing or rising, and dam digging or burrowing in shavings. The VN dams and their litters were not significantly different from the control dams and their litters on any of the measures taken. The ZN dams and their litters were not significantly different from their controls on all measures except for pup body temperature which was slightly lower from Day 13 through day 16 and pup body weight which was slightly lower from Day 12 through Day 16. Since these differences are very small, they do not seem to indicate a serious deficiency in maternal behavior. The results indicate that adequate maternal behavior develops with either of the two systems intact.     

Olfactory basis for the delayed onset of maternal behavior in virgin female rats: experimential effects.

A previous study showed that intranasal zinc sulfate (ZnSO4) treatment shortens the latencies for the onset of maternal behavior in nonpregnant rats. The aim of this study was to see whether after recovery from the effects of ZnSO4 treatment, the latencies for maternal behavior increased. In Experiment 1 nonpregnant females were intranasally infused with ZnSO4 or air or were left untreated and exposed to pups starting 48 hr later. Olfactory discrimination tests using chocolate bits were done simultaneously. The ZnSO4 treatment resulted in short latency maternal behavior (1.1 days) compared with latencies of control females (3.8 days), but neither group showed any loss of olfactory discrimination. Three weeks later- ZnSO4-treated females showed increased latencies for all but retrieving, while control groups showed decreased latencies for all maternal behaviors. In Experiment 2, intranasal ZnSO4- and air-treated females were given olfactory discrimination tests under food deprivation, using chocolate bits and guinea pigs pellets. Choice of guinea pig pellets was more severely affected than choice of chocolate bits, but recovery of the discriminations was complete in 4-5 days. The effects of ZnSO4 on maternal behavior and on olfactory discrimination in nonpregnant females were discussed.     

Chronic exposure to a novel odor increases pups' vocalizations, maternal care, and alters dopaminergic functioning in developing mice

This study was designed to assess the stress effect of manipulation of the olfactory environment in developing mice. In a first experiment it was found that mouse pups could be stressed (as measured by an increase in ultrasonic calls) by removing the litter from the dam for 15 min/day for the first 14 days of life and exposing them to a novel odor (clean bedding). This stress procedure also produced a long-term modification in maternal behavior. The stress response (ultrasounds) and the modification of maternal behavior were prevented by providing the litter with home cage bedding during maternal separation. In a second experiment it was demonstrated that early stress influenced apomorphine-induced wall climbing behavior in 15-day-old mice, suggesting stress-induced alterations in the dopaminergic system. Pups exposed to clean bedding during infancy exhibited more wall climbing behavior than pups never separated from the mother. Moreover, preventing the early stress response during mother-offspring separation, by providing pups with home cage bedding, eliminated the increase in apomorphine-induced wall climbing. Taken together these results suggest that olfactory cues are decisive in characterizing stressful situations inducing both immediate and long-lasting effects in mouse pups.     

Decline of maternal behavior in the virgin and lactating rat.

Sensitized virgins and postpartum lactating mothers, both exhibiting maternal behavior, were given donor litters that increased in age by 1 day, for 28 days, starting at the onset of maternal behavior. Each day females were tested for maternal behavior with pups 4-8 days old: Maternal care (i.e., nursing/crouching, retrieving, nest building and licking) and maternal withdrawal, rejection, and prevention of nursing were recorded. After the ninth day, females were also tested with the progressively older pups from 10 to 28 days of age with which they were living. Virgins and lactating mothers showed generally similar patterns of maternal care although some differences were found, and they declined in maternal behavior toward the older pups in a similar manner. Maternal behavior did not decline in tests with younger pups. The results are interpreted as supporting the hypothesis that the decline as well as the maintenance of maternal behavior postpartum is nonhormonally mediated.     

Analysis of the catnip reaction: mediation by olfactory system, not vomeronasal organ

Pet owners and behavioral scientists alike are fascinated by unique behavioral reactions that cats show in the presence of catnip. These experiments explored the possibility that the catnip reaction might be triggered by chemosensory stimulation of the vomeronasal organ. In the chewing and mouthing of the catnip source, substances might be dissolved in saliva and transported to the vomeronasal organ. The rolling and rubbing during a catnip reaction might be a sexual response activated by the accessory olfactory system since the system projects to parts of the brain involved in mediation of sexual behavior. However, removal of the vomeronasal organ did not attenuate any of the behavioral reactions to catnip. Olfactory bulbectomy immediately eliminated catnip responding, revealing that the chemosensory stimulus evoking the catnip reaction is undoubtedly mediated through the main olfactory system. Catnip activates behavioral elements associated with several species-specific behaviors, including sniffing and chewing as associated with oral appetitive behavior, rolling and rubbing characteristic of female sexual behavior, batting the catnip source characteristic of play behavior, and a type of kicking associated with predatory behavior. These behavioral reactions occur randomly and intermittently.     

The development of olfactory preferences for artificial odors briefly experienced by the precocial spiny mouse young

Groups of pups, aged 2, 4, 6, 12, 16, and 18 days of the precocial murid rodent spiny mouse (Acomys cahirinus), were each exposed for 1.5 h to one of the experimental odors, cinnamon or cumin, present in the surrounding environment. The odors were not contingent upon home-cage odors or any other reinforcers. This simple exposure produced a strong preference for the exposed odor in multichoice preference tests carried out 48 h after the exposure in pups from groups aged 2 through 16 days. Older pups exposed to cinnamon or cumin at Day 18 did not show a significant preference for the familiar odor. Both experimental odors used were found to be neutral for control experimentally naive pups within a tested age span. These results point at a stimulus familiarity, rather than associative learning in the development of olfactory preferences in young spiny mouse pups.     

Olfactory stimulation induces filial preferences for huddling in rat pups.

Rat pups of all ages huddle with conspecifics, but the senosory control of contact behavior changes ontogenetically. Thermal cues control huddling until about Day 15, at which time species' odors become the dominant stimulus. The present experiments indicate that the filial response to conspecifics is dependent on olfactory experience. A synthetic chemical scent was added to the smells of the dam from Day 1 to Day 20 postpartum. Standardized videographic tests were used to assess the development of huddling preference. Preferences for nest-typical smells emerged by Day 15 in pups from both scented and nonscented litters. Pups from scented nests preferred to huddle with a scented stimulus rat, whereas control pups preferred a nonadulterated rat stimulus. Additional testing indicated that the affiliative preferences were specific to rearing odor and were not based on decreased aversion to test scents, or on disrupted olfactory discrimination. The ontogeny of species-typical contact behavior is discussed in terms of the induction of a perceptual preference that is based on early odor stimulation.     

Long-term effects of preoptic area knife cuts on the maternal behavior of postpartum rats

Knife cuts severing the lateral connections of the medial preoptic area disrupt maternal behavior in postpartum rats. In these studies the knife cuts were produced 1-2 days prior to behavioral testing. The present study examined whether preoptic area knife cuts produce long-term deficits in maternal behavior. Nulliparous Charles River CD rats received knife cuts severing the lateral connections of the medial preoptic area or sham cuts. Approximately 2 weeks later they were mated. All females delivered young 22-23 days after mating, and litter size did not differ between the groups. Females with preoptic knife cuts cleaned their young, built nests (albeit of inferior quality), crouched over their young in a nursing posture, and lactated. The major deficit shown was the near total elimination of retrieval behavior. Females with preoptic knife cuts, however, were capable of performing the oral-motor responses associated with hoarding behavior. These results show that severing the lateral connections of the medial preoptic area does produce long-term deficits in maternal behavior, particularly with respect to maternal retrieving.     

Sex differences in maternal behaviors of immature rats: The role of emotionality

Immature rats are able to give maternal care to pups and may increase their inclusive fitness by helping newborn siblings. Immature males are generally reported to be more maternal than females. In this research we studied sex differences and the extent to which they may be due to a different emotional response to novelty. We studied maternal response in two groups of immature rats of both sexes: in the naive group, juveniles aged from 18 to 30 days were given one test and showed clear-cut sex differences. The experience group was submitted to repeated tests from 18 to 30 days of age, thus eliminating the effects of novelty: as a result, all differences between sexes disappeared. A second experiment was designed in 24-day-old females to differentiate the effects of repeated handling and exposure to the test situation from that of repeated exposure to pups; moreover, the effects of an anxiety-reducing drug (diazepam) were studied. Exposure to pups is particularly efficient in facilitating maternal response. Diazepam injection was also effective, but its control showed that the injection per se was responsible for the effect. In conclusion, our results stress the importance of response to novelty and suggest that sex differences in maternal behavior of immature rats may be modulated by different emotional responses by males and females. In fact, emotionality competes with, and may exert an inhibitory effect on, the expression of a vast array of behaviors. © 1994 Wiley-Liss, Inc.     

Prepartum changes in maternal responsiveness and nest defense in Rattus norvegicus

Changes in maternal responsiveness during late pregnancy were traced by exposing females to foster pups (15-30-min tests) under conditions favoring the rapid initiation of maternal behavior. Groups were tested when nonpregnant or on Days 17, 20, 21, or 22 of pregnancy (Day 22 = parturition). Nulliparous females were compared with primiparous and with "experienced breeders." Nest defense was observed by introducing unfamiliar males (2-min tests) on Day 22. Results focused attention on three periods: (a) Days 17-20, when maternal responsiveness was lower than in the nonpregnant condition, (b) Day 21, when maternal responsiveness returned to or rose above nonpregnant levels, and (c) on Day 22, the 3.5 hr prior to delivery, during which 90% of females almost immediately retrieved, gathered, and tended foster pups and during which 92% attacked the unfamiliar intruders. (Attacks were rare earlier.) Maternally experienced females were more responsive to pups than nulliparous females when nonpregnant and throughout late pregnancy, but both groups were equally likely to show prepartum aggression.     

Interactions between perinatal and neonatal associative learning defined by contiguous olfactory and tactile stimulation

Tactile stimulation of the neonate, as performed by the mother during and after delivery, has been described as an effective unconditioned stimulus during early ontogeny (Leon, 1987; Ronca & Alberts, 1994). The present experiments examined the interaction between perinatal and neonatal learning determined by the explicit association between alcohol odor and vigorous body stimulation of the perinatal organism. In Experiment 1, rat fetuses were exposed to either alcohol or saline 10 min prior to cesarean delivery. The alcohol administration procedure here employed was sufficient to provide sensory contamination of the amniotic fluid but avoid fetal alcohol intoxication. Pups in the two prenatal treatments later experienced the smell of alcohol, tactile stimulation, or both stimuli explicitly paired or unpaired. Other postnatal groups were composed of pups that had no explicit experience with either experimental stimulus. Pups subjected to alcohol odor in utero displayed more overall motor activity in response to that odor than saline controls. The increased motor responses were further potentiated in pups that experienced additional postnatal alcohol odor paired with tactile stimulation. In Experiment 2, pups were exposed to alcohol in the amniotic fluid 10 or 30 min prior to birth. As previously demonstrated the memory acquired in utero appears highly dependent upon contingency between exposure to this particular scent and delivery procedures. Pups in both prenatal treatment groups were then exposed to alcohol odor paired or unpaired with tactile stimulation. Some control animals received no further experience with either stimuli. Those pups exposed to alcohol odor paired with tactile stimulation both pre- and postnatally later showed maximum motor activity elicited by the odor of alcohol. The results support the notion of fetal associative learning comprising alcohol's chemosensory cues and behaviorally activating stimuli. Furthermore, the conditioned response under analysis is potentiated whenever neonates are reexposed to contingent presentations of the elements that defined the original associative memory.     

Stimulus control of maternal responsiveness to Norway rat (Rattus norvegicus) pup ultrasonic vocalizations

Mother rats (Rattus norvegicus; 6 to 8 days postpartum) approach and maintain proximal orientation to a pup that is emitting ultrasonic vocalizations (USVs) far more than do virgin females (W. J. Farrell & J. R. Alberts, 2002). We used a playback regimen to examine the roles of acoustic and nonacoustic cues in regulating maternal proximal orientation toward vocalizing pups. When presented with recorded USVs, mothers of 6- to 8-day-old pups and nulliparous virgin females exhibited equivalent levels of proximal orientation toward the playback speaker. Mothers did show enhanced proximal orientation toward recorded USVs, however, if a silent pup was positioned below the speaker. Pup odors appear to be crucial for the maternal response to vocalizing pups, as peripherally induced anosmia attenuated maternal proximal orientation toward a vocalizing pup. Furthermore, spatial contiguity between olfactory and auditory stimuli was required for a maximal maternal response.     

Nonselective maternal bonding but pup recognition in the subterranean rodent Ctenomys pearsoni

This study characterizes the maternal behavior in the subterranean rodent Ctenomys pearsoni and determines whether lactating females develop a selective bonding to their own pups in addition to maternal responsiveness. Mother-litter interactions were observed daily for 2 months; during the 1st postpartum week, selectivity and preference tests for own or alien pups were performed. Lactating females exhibited all patterns of maternal behavior observed in other rodent species and equally responded with care-taking behaviors to their own and alien pups, although they were able to discriminate between them, showing preference for their own pups. Results are discussed, taking into account the possible role of this behavior in shaping different aspects of life underground in ctenomyid species.     

Maternal behavior of spontaneously hypertensive and Wistar-Kyoto normotensive rats: effects of reciprocal cross-fostering of litters

Maternal behavior of spontaneously hypertensive (SHR) and Wistar-Kyoto (WKY) normotensive dams rearing either their natural pups or pups of the opposite strain was assessed when pups were 4-6 days of age. Consistent with previous reports, SHR dams caring for their natural pups were observed nursing and licking their pups more often than WKY dams and were away from their pups less often. The behavior of dams rearing cross-fostered pups demonstrated that this strain difference in patterns of maternal behavior is dependent on pup type. Mothers of both strains shifted their behavior in the direction of the opposite strain's maternal behavior pattern when caring for cross-fostered pups. This shift was most pronounced in SHR dams caring for WKY pups. These data suggest that alterations in patterns of maternal behavior produced by cross-fostering may play an important role in cross-fostering effects on the physiological development of SHRs previously reported.     

Hysterectomy-induced maternal behavior during pregnancy in the rat.

Hysterectomies were performed between the 10th and 19th day of pregnancy, and females were tested with pups for the onset of maternal behavior starting 0,24, 48, or 72 hr after surgery. Pups remained with the females overnight, and testing was repeated daily with fresh pups until females exhibited maternal behavior. Latencies for the onset of maternal behavior were shorter after hysterectomy on the 10th and 16th days of pregnancy than in intact pregnant females at the same stages of pregnancy; latencies became shorter, the later the termination of pregnancy. When the ovaries were removed along with hysterectomy during pregnancy, short-latency maternal behavior no longer was exhibited. Pregnant females were tested during the last 40 hr of pregnancy: nest building began at 34 hr prepartum and retrieving at 28 hr prepartum. The effect of hysterectomy during pregnancy on ovarian secretion of estrogen and progesterone is reviewed, and it is concluded that the rise in estrogen secretion, which follows hysterectomy during pregnancy, is most likely the cause of the rapid onset of maternal behavior after hysterectomy. A similar proposal is made for the prepartum onset of maternal behavior in intact pregnant females.     

Incidence of pup killing and parental behavior in virgin female and male rats (Rattus norvegicus): differences between Wistar and Sprague-Dawley stocks

The responses toward young shown by males and nulliparous females differed substantially between two outbred stocks of laboratory rats. Sprague-Dawley females showed maternal behavior either spontaneously (35% of the naive rats) or through concaveation (92% of the initially neutral virgins). Of the Wistar females, however, only 10% showed maternal behavior spontaneously, and only 29% of the neutral virgins came to behave maternally during 15 days of concaveation. Prepubertal cohabitation with lactating rats did not facilitate maternal responsiveness in adulthood in the Wistar virgin females. Of the Sprague-Dawley males, 50% showed paternal behavior spontaneously, and only 4% killed the young. Among the Wistar males, however, only 4% showed paternal behavior spontaneously, and 76% killed pups. Such profound differences between outbred stocks of rats may be a source of discrepancies between the results of studies dealing with the induction of parental behavior in nonlactating rats.     

Differential Effects of Short- and Long-Term Early Maternal Separation on Subsequent Maternal Behavior in Rats

Female Sprague-Dawley pups were separated from mothers every other day for 8 hr (long-term separation/LTS), 4 hr (short-term separation/STS), or 0 hr (no separation/NS) from postnatal day 2–20. In adulthood, they were mated and tested for maternal behaviors during two lactations. It was expected that females separated from mothers as pups would show deficits in maternal behavior as adults. Contrary to expectations, LTS showed better nest building and grouped young faster during both lactations. LTS were first to display aggression and displayed more aggression during the second lactation. Notably, while some measures decreased from first to second lactation in NS and STS, LTS maintained levels of maternal care. These results suggest that extended periods of maternal separation may exaggerate some aspects of maternal behavior.     

Association between chemosensory stimuli and cesarean delivery in rat fetuses: neonatal presentation of similar stimuli increases motor activity

Recent studies conducted in this laboratory indicated that prenatal chemosensory stimulation followed by cesarean delivery strongly affected postnatal responsiveness to odors derived from the administered substances. The present experiments were performed in order to examine if an associative process was responsible for such effects. In Experiment 1 rat fetuses during Gestational Day 21 were exposed to a tenuous alcohol solution or to a lemon-containing solution either 40 or 10 min prior to cesarean delivery. All subjects were subsequently tested in terms of changes in neonatal motor activity when confronted with the odor of alcohol or lemon. Rats experiencing prenatal cues 10 min prior to delivery exhibited higher and differential responsiveness to the smell of these cues when compared to those experiencing similar solutions 40 min prior to delivery. In Experiment 2 each fetus sequentially experienced both cues. Subsequent tests confirmed that the delay between prenatal sensory experience and birth induction was critical in terms of significantly affecting olfactory-mediated motor responses. The results suggest that consequences related with cesarean delivery act as an unconditioned stimulus capable of being associated with orosensory cues present in the amniotic fluid.