Maternal exercise decreases maternal deprivation induced anxiety of pups and correlates to increased prefrontal cortex BDNF and VEGF

Maternal deprivation (MD) may cause neuropsychiatric disorders such as anxiety disorder by negatively affecting the cognitive functions and behavior in pups. The aim of this study is to investigate whether maternal exercise during pregnancy has beneficial effects on anxiety that increases with MD, and on the levels of VEGF and BDNF which have anxiolytic effects on the prefrontal cortex, the anxiety-related region of the brain. The anxiety level in the deprivation group was greater than the control group and found more in male than female pups. The prefrontal cortex VEGF and BDNF levels were decreased in the deprivation group compared to control group while serum corticosterone levels were increased in the deprivation group. Anxiety and serum corticosterone levels were decreased in maternally exercised female and male pups, while the prefrontal cortex VEGF and BDNF levels were increased, compared to sedentary mother's pups. These results indicate that maternal exercise may attenuate the negative effect of stresses such as maternal deprivation that can be encountered early in life.     

Some effects of a split litter cross foster design applied to 15 day old rat pups

Rat litters were divided and cross fostered on a postnatal Day 15 so that each mother had 4 of her own pups and 4 pups of a second litter, until weaning at 21 days. Daily weight and body temperature measurements made between Day 15 and 21 showed no differences between foster pups and pups who stayed with their biological mothers. However, by the age of 30 days, foster pups weighed less than pups reared by their own (biological) mothers; and foster pups at that age did not survive food deprivation as well as the pups reared by their biological mothers. In their Day 30 weights and the capacity to survive food deprivation, foster pups resembled pups that had been permanently separated from their mothers on postnatal Day 15. We conclude that the split cross foster design may introduce variability rather than reduce it. Nonetheless, this design may be useful in the experimental investigation of maternal behavior and mother-pup interactions in the rat.     

Sleep deprivation effects on growth factor expression in neonatal rats: a potential role for BDNF in the mediation of delta power

The sleeping brain differs from the waking brain in its electrophysiological and molecular properties, including the expression of growth factors and immediate early genes (IEG). Sleep architecture and homeostatic regulation of sleep in neonates is distinct from that of adults. Hence, the present study addressed the question whether the unique homeostatic response to sleep deprivation in neonates is reflected in mRNA expression of the IEG cFos, brain-derived nerve growth factor (BDNF), and basic fibroblast growth factor (FGF2) in the cortex. As sleep deprivation is stressful to developing rats, we also investigated whether the increased levels of corticosterone would affect the expression of growth factors in the hippocampus, known to be sensitive to glucocorticoid levels. At postnatal days 16, 20, and 24, rats were subjected to sleep deprivation, maternal separation without sleep deprivation, sleep deprivation with 2 h recovery sleep, or no intervention. mRNA expression was quantified in the cortex and hippocampus. cFos was increased after sleep deprivation and was similar to control level after 2 h recovery sleep irrespective of age or brain region. BDNF was increased by sleep deprivation in the cortex at P20 and P24 and only at P24 in the hippocampus. FGF2 increased during recovery sleep at all ages in both brain regions. We conclude that cortical BDNF expression reflects the onset of adult sleep-homeostatic response, whereas the profile of expression of both growth factors suggests a trophic effect of mild sleep deprivation.     

Maternal deprivation in neonatal rats of different conditions affects growth rate, circadian clock, and stress responsiveness differentially

Effects of periodic maternal deprivation (MD) were examined in rat pups on growth rate, circadian phase and period at weaning, and stress responsiveness in adulthood. MD was performed from postnatal day 1 to day 6 or day 7, with or without keeping ambient temperature at 37 degrees C and humidity at 70-80% during deprivation. Times of day and length of MD were also changed. Body weights were significantly reduced at weaning in MD12 (MD for 12 h) and MD6am (MD for 6 h in the morning) pups, whereas they were not changed in MD6pm (MD in the afternoon) and all MD3 groups. At 8 weeks old, body weight was still significantly lower in MD12 than the control, but not different from the control in other groups. Circadian phases of free-running locomotor rhythm at weaning were almost reversed in MD12, MD6am and MD6pm as compared with those in the control. Intermediate phase-shifts were observed in MD3Eam (3 h MD in the first quarter of the light phase; early am) and MD3Lam (late am; the second quarter), whereas no phase-shift was detected in MD3Epm (early pm; the third quarter) and MD3Lpm (late pm; the fourth quarter). Elevation of plasma corticosterone level after novelty exposure at 8 weeks old was more robustly in MD12 and MD3Lam than in the control, but the hormone response in MD3Lpm was not different from the control. Keeping ambient temperature at 37 degrees C during MD did not rescue the MD-induced body weight loss, but attenuated the phase-shifts of the circadian clock, and completely cancelled the stress-induced hormone response in MD12 rats. These findings indicate that MD in rat pups differentially affects growth rate, circadian clock, and stress responsiveness in adulthood, depending on time of day, length of MD and ambient temperature during MD.     

Ontogeny of feeding controls in suckling and weanling rats.

To rationalize conflicting data in the literature, the latency of attachment of fed and fasted rat pups to the nipples of their anesthetized mother has been studied at various postnatal ages. At 10 and 14 days of age, the mode of fasting had a critical effect on the results: pups fasted in a 30 degrees C incubator had shorter attachment latencies than fed pups, whereas when pups were fasted in a 34 degrees C incubator or with a nonlactating foster mother at room temperature, their latencies were no different from those of fed pups. Thus maternal deprivation has an important influence on attachment latencies, unless other steps are taken to maintain the body temperature of fasted pups. When this variable is controlled, it is apparent that at 10 and 14 days of age, the feeding behavior of rat pups is not related to nutritional status. In contrast, from 17 days onward, latencies of fed pups were significantly longer than those of faster pups, regardless of the mode of fasting. This appearance of appetitive behavior during the 3rd postnatal wk was shown to have a temporal correlation with the beginning of weaning, as measured by the appearance of chow in the stomachs.     

Maternal deprivation potentiates pituitary-adrenal stress responses in infant rats

In three experiments we examined the effect of maternal deprivation on the pituitary-adrenal response of 12-, 16- and 20-day-old rat pups to novelty stress. Infant rats were either deprived individually in heated incubators or left in the home nest with their mother and then tested for their corticosteroid response to 30-min exposure to a novel test arena (novelty-stress). In Experiment 1 we showed that the magnitude of the stress response was a positively accelerated function of the deprivation interval. Stress responses were not increased after 1 hour of deprivation, were modestly increased after 8 hours of deprivation, and were dramatically increased after 24 hours of deprivation. In Experiment 2 we asked whether potentiation of the stress response resulted from the maternal or the nutritive components of the deprivation procedure. Pups were tested under one of four treatment conditions formed by a 2 (Maternally Deprived vs. Nondeprived) x 2 (Nutritively Deprived vs. Nondeprived) factorial design. At 12 and 16 days of age, potentiation of the stress response was traced to the absence of maternal care and not nutrients. At 20 days of age, both maternal and nutritive deprivation contributed to the potentiated stress response. The results of Experiment 3 showed that this effect was mediated, at least in part, by increased adrenocortical sensitivity to ACTH, because the corticosteroid response to exogenous ACTH administration was also increased by maternal deprivation. These findings add to a growing body of literature that supports the concept of maternal regulation of infant physiology. They also support previous reports from this laboratory indicating that suppression of the pituitary-adrenal system is modulated by maternal variables during the preweaning period in the rat.     

Long-term consequences of early maternal deprivation in serotonergic activity and HPA function in adult rat

Increasing body of evidence indicates that early life stressful events may induce permanent alterations in neurodevelopment, which in turn, could lead to the development of psychopathologies in adulthood. In particular, maternal deprivation (MD) for 24 h in rats has been associated with several abnormalities in brain and behaviour during adulthood, relevant to the neurobiological substrate of anxiety disorders. The aim of the present study was to clarify the long-term effects of MD, on hypothalamo–pituitary–adrenal (HPA) axis activity and serotonergic (5-HT) function, in adulthood, subjects that have not been yet thoroughly investigated. For this purpose, Wistar rat pups were deprived from their mothers for a 24-h single period at postnatal day 9 (pnd 9) and were examined when aged 69–90 days. Plasma corticosterone and ACTH levels along with the animal's behaviour in an open field were used as indices of stress. Moreover, serotonergic activity was estimated in hypothalamus and hippocampus, key structures in the coordination of neuroendocrine and behavioural responses to stress. Interestingly, in adulthood, MD rats compared to controls, displayed decreased body weight, increased serotonergic activity and “anxiety” related behaviour, as well as elevated plasma corticosterone and ACTH levels. The findings of this study showed that MD results in long-term modifications in HPA axis and serotonergic activity indicating a clear relationship between early life stressful events and the development of anxiety-like disorders later in adulthood.     

Maternal treadmill exercise during pregnancy decreases anxiety and increases prefrontal cortex VEGF and BDNF levels of rat pups in early and late periods of life

In a previous study we demonstrated that, regular aerobic exercise during pregnancy decreased maternal deprivation induced anxiety. The purpose of this study is to investigate whether the positive effects of maternal exercise on the male and female offspring's early and late period of life. Half of the test subjects in each group were evaluated when they were 26 days old, and the other half were evaluated when they were 4 months old. The anxiety levels of maternally exercised groups were less than the control groups in both sexes and in both prepubertal and adult periods. The locomotor activity more increased in females. The prefrontal VEGF and BDNF levels were greater for both age groups and sexes in the maternally exercised group compared to control group. Moreover, there was a strong positive correlations between prefrontal cortex BDNF levels and results of open field tests; and VEGF levels and results of elevated plus maze tests. There was no difference in serum corticosterone levels between groups. These results indicate that maternal exercise during pregnancy may protect the pups from anxiety in early and late periods of life, and affects the prefrontal cortex positively.     

The ontogeny of independent ingestion in mice: Or,why won't infant mice feed?

Unlike infant rats, which show deprivation-related ingestion in several different test situations, infant mice appeared to be relatively unwilling to feed independently of suckling until 12 days of age. We tested mouse pups that were deprived (of food, water, suckling, and maternal care) for 1, 7, or 24 hr in ingestive tests in which a milk diet was spread on the floor of their test container (Experiment 1). Pups at 3, 6, and 9 days of age consumed small amounts of the diet and showed little increase in intake when deprivation was increased. In contrast (and like rat pups of all ages), mouse pups 12 and 15 days of age actively ingested the diet and increased their intake with increased deprivation. Six-day-old mouse pups were similarly unwilling to ingest a 5% sucrose solution, though 12-day-old pups showed deprivation-related intake (Experiment 2). Cellular dehydration (produced by hypertonic saline injection), a potent stimulus for ingestion in infant rats, did not stimulate ingestion in mice younger than 12 days of age (Experiment 3). Finally, when ingestion was tested with diet infusions made through oral cannulas, mouse pups at 6 and 9 days of age showed only a slight increase in intake with increased deprivation. However, by 12 days of age, pups' ingestion increased markedly with deprivation (Experiment 4). Thus, mouse pups seem to be very different from rat pups with respect to the early existence of ingestive systems. The neural substrates for the ingestive responses that subserve independent ingestion are only minimally present in infant mice or are somehow inhibited.     

Kindling in the early postnatal period: Effects on the dynamics of age-related changes in electrophysiological characteristics of hippocampal neurons

The effects of chronic administration of pentylenetetrazole (PTZ) during early ontogenesis (from postnatal day 14) on the dynamics of age-related changes in electrophysiological characteristics of rat hippocampal slices were studied. Unlike the situation in adult animals, convulsive activity did not develop in rat pups in response to repeated injections. Comparison of the amplitude characteristics of total monosynaptic responses of neurons in hippocampal field CA1 to application of single and paired (separated by 70 msec) stimulation of Schäffer collaterals during the period of maximally intense hippocampal synaptogenesis (at weeks 2–3 of postnatal development) revealed no significant differences between the control group (administration of isotonic saline) and the group given PTZ. The level of suppression of facilitation in paired-pulse stimulation with a short interstimulus interval (15 msec) was significantly less in hippocampal slices from rat pups from the PTZ group. However, as compared with the passive control, the direction of rearrangements in the two experimental group was essentially the same. Nonetheless, regular administration of PTZ during the period of maximally intense hippocampal maturation affected the development of its characteristics. This was not only apparent as a deficiency of inhibitory processes. Increases in the intensity of test stimuli applied to hippocampal slices from PTZ-treated rat pups at 27–48 days of age led to relatively lower response amplitudes as compared with those seen in hippocampal slices from control (given isotonic saline) rats of the same age. The level of facilitation in paired-pulse stimulation with an interstimulus interval of 70 msec showed no difference, decreasing to similar extents in both groups as compared with the passive control group. In addition, hippocampal slices from the PTZ group showed significant decreases in the magnitude of long-term potentiation. Changes occurring in the hippocampus after regular administration of PTZ did not correlate with the development of convulsive activity. The only significant relationship involving the intensity of convulsions was with the increase (compared with the normal for age) in the amplitudes of responses to minimal-intensity test stimuli.     

Acupuncture stimulation at HT7 alleviates depression-induced behavioral changes via regulation of the serotonin system in the prefrontal cortex of maternally-separated rat pups

A possible application of acupuncture in alleviating depression-like behavioral changes and regulating serotonin signaling in the prefrontal cortex (PFC) of maternally-separated rat pups was investigated in this study. On postnatal day 15, rat pups were maternally-separated and received acupuncture stimulation at acupoint HT7 or ST36 once a day for 7 days. On postnatal day 21, the tail suspension test was performed and the PFC was harvested. Tissue levels of serotonin (5-HT) and 5-hydroxyindole-3-acetic acid (5-HIAA) were then measured by high-performance liquid chromatography and expression of serotonin transporter (5-HTT) and brain-derived neurotrophic factor (BDNF) were assessed by western blotting. Levels of 5-HT and 5-HIAA were not significantly changed, but the 5-HIAA/5-HT ratio was significantly increased by maternal separation. The immobility time of maternally-separated rat pups was increased, and increased 5-HTT expression and reduced BDNF level were observed in the PFC. But acupuncture stimulation at HT7 alleviated the behavioral change and regulated the changes of 5-HIAA/5-HT ratio, 5-HTT, and BDNF. In conclusion, acupuncture stimulation at HT7 can relieve maternal separation-induced changes, and we propose that regulation of the 5-HIAA/5-HT ratio and of 5-HTT expression by acupuncture stimulation are important acupuncture-induced benefits in this animal model of depression.     

Maternal separation impairs long term-potentiation in CA1-CA3 synapses and hippocampal-dependent memory in old rats

Exposure to chronic stress during the neonatal period is known to induce permanent long-term changes in the central nervous system and hipothalamic-pituitary-adrenal axis reactivity that are associated with increased levels of depression, anxiety, and cognitive impairments. In rodents, a validated model of early life stress is the maternal separation (MS) paradigm, which has been shown to have long-term consequences for the pups that span to adulthood. We hypothesized that the early life stress-associated effects could be exacerbated with aging, because it is often accompanied by cognitive decline. Using a MS model in which rat pups were separated from their mothers for 3 hours daily, during postnatal days 2–14, we evaluated the long-term functional consequences to aged animals (70-week-old), by measuring synaptic plasticity and cognitive performance. The baseline behavioral deficits of aged control rats were further exacerbated in MS animals, indicating that early-life stress induces sustained changes in anxiety-like behavior and hippocampal-dependent memory that are maintained much later in life. We then investigated whether these differences are linked to impaired function of hippocampal neurons by recording hippocampal long-term potentiation from Schaffer collaterals/CA1 synapses. The magnitude of the hippocampal long-term potentiation induced by high-frequency stimulation was significantly lower in aged MS animals than in age-matched controls. These results substantiate the hypothesis that the neuronal and endocrine alterations induced by early-life stress are long lasting, and are able to exacerbate the mild age-associated deficits.     

Patterned (single) alternation in infant rats after combined or separate lesions of hippocampus and amygdala

The role of the developing hippocampus and the amygdala on patterned (single) alternation (PA) in the infant rat was investigated in 4 experiments. In Experiments 1 and 2, pups were given 2 bilateral electrolytic hippocampal lesions or sham surgeries at 10 or 11 days of age and were trained 6 days later in a straight runway. In Experiment 1, there were 120 trials in 1 day, with an 8-, a 15-, or a 30-s intertrial interval (ITI). PA learning occurred in lesion and sham pups at the 8- and 15-s ITIs, but it was reduced in both groups at the 30-s ITI. In Experiment 2, training was extended to 240 trials over 2 days, with a 30- or 60-s ITI. Sham and lesion pups showed PA at the 30-s ITI, but the emergence of PA was delayed in the lesion pups at the 60-s ITI. In Experiment 3, amygdaloid lesions had no effect on PA learning at the 8-s ITI. However, when pups with hippocampal and amygdaloid lesions were trained at the 8-s ITI, the emergence of PA was delayed, and its size was reduced (Experiment 4). The results of these experiments argue for a role of the hippocampus in PA learning at long ITIs and suggest that, even in 16-day-old pups exposed to an 8-s ITI, the combined hippocampal and amygdaloid lesion produces a deficit greater than either the hippocampal or the amygdaloid lesion. The results are discussed in relation to current theories that distinguish between 2 levels of memory function.     

Prenatal stress produces sex differences in nest odor preference

Prenatal stress (PS) and early postnatal environment may alter maternal care. Infant rats learn to identify their mother through the association between maternal care and familiar odors. Female Wistar rats were exposed to restraint stress for 30 min, 4 sessions per day, in the last 7 days of pregnancy. At birth, pups were cross-fostered and assigned to the following groups: prenatal non-stressed mothers raising non-stressed pups (NS:NS), prenatal stressed mothers raising non-stressed pups (S:NS), prenatal non-stressed mothers raising stressed pups (NS:S), prenatal stressed mothers raising stressed pups (S:S). Maternal behaviors were assessed during 6 postpartum days. On postnatal day (PND) 7, the behavior of male and female pups was analyzed in the odor preference test; and noradrenaline (NA) activity in olfactory bulb (OB) was measured. The results showed that restraint stress increased plasma levels of corticosterone on gestational day 15. After parturition, PS reduced maternal care, decreasing licking the pups and increasing frequency outside the nest. Female pups from the NS:S, S:NS, S:S groups and male pups from the S:S group showed no nest odor preference. Thus, at day 7, female pups that were submitted to perinatal interventions showed more impairment in the nest odor preference test than male pups. No changes were detected in the NA activity in the OB. In conclusion, repeated restraint stress during the last week of gestation reduces maternal care and reduces preference for a familiar odor in rat pups in a sex-specific manner.     

Periodic exposure to mother is potent zeitgeber of rat pups' rhythm

To study the possibility and potency of periodic maternal deprivation (PMD) in entraining the circadian rhythm of rat pups, access to the natural mother of which pups were optically enucleated on the day of birth (day 1) was restricted to either light phase (L-group) or dark phase (D-group) for various durations beginning on various days during the nursing period. Drinking rhythm of both groups was determined once per one or two weeks between the 4th and 8th postnatal week. The corticosterone rhythm was determined to confirm the results obtained by the measurement of water intake. Both rhythms were clearly observed by 5 weeks of age, and a reversed phase relationship was observed even when PMD was performed for a short period during days 1-4 or days 15-21. On the other hand, no phase angle difference was observed between L- and D-groups, when pups were periodically exposed to their original mother either during days 1-2 or days 18-21. These facts indicate that periodic exposure to mother can set the phase of the blinded pups' rhythm and that the period required for entrainment of the rhythm is as short as 4 or 7 days in the early or late times during the nursing period respectively, showing the high potency of PMD in entraining the circadian rhythm of blinded pups.     

Adrenocortical response to stress in fasted and unfasted artificially reared 12-day-old rat pups

Recent studies have compared artificially reared (AR) rats with dam-reared rats on behavioral outcomes but, despite the fact that they are deprived of their mothers during the stress hyporesponsive period (SHRP), little is known about the effects of AR on the development of the stress response. In this study, the corticosterone (CORT) response to a stressor (saline injection ip) on postnatal Day 12 was assessed in rat pups that had been either dam-reared (DR) or artificially reared since Day 5. In the preceding 24 hr, half the pups in the DR group were maternally deprived (DEP). To control for the food deprivation consequent to maternal deprivation in these groups, half the pups in the AR groups also underwent 24-hr food deprivation (DEP). In the nondeprived condition AR pups did not differ from DR pups on untreated CORT levels or on levels at 1-hr poststress (i.e., all rats demonstrated low levels of CORT characteristic of the SHRP). In contrast, both maternally deprived DR pups and food-deprived AR pups exhibited increased untreated CORT levels as well as a significant increase at 30-min poststress, but CORT elevations were lower in the AR groups than in the DR groups. Thus, long-term maternal deprivation through artificial rearing in rats does not affect the reduced CORT levels and reduced CORT responsiveness associated with the SHRP; however, if animals are food deprived, then all show increased basal CORT levels and a greater CORT response to stress, although this response is lower in AR groups than in DR groups. These results suggest that rat pups artificially reared with adequate nutrition will still exhibit the SHRP.     

Behavioral and physiological effects of early handling and early malnutrition in rats.

This experiment employed a 2 x 2 design in which the nutritional and early stimulation conditions of rat pups were manipulated during the 1st 3 weeks of postnatal life. Animals were observed in various open field activities at 22, 23, 29, 36, and 50 days of age, and killed at 55 days of age for neurochemical analysis. Early handling decreased the behavioral differences due to malnutrition and resulted in fewer, but larger cells in rat forebrain for both well-nourished and malnourished groups.     

Early weaning decreases play-fighting behavior during the postweaning developmental period of Wistar rats

We examined the influence of early weaning on the development of play-fighting behaviors and anxiety status in Wistar rats. Pups were divided into two groups, those weaned at postnatal day (PD) 16 (early-weaned group) and those weaned at PD30 (normally weaned group), and were housed in pairs of the same sex. Playful interactions were measured for each pair once a week from 4 to 7 weeks of age. Thereafter, during early adulthood, all the rats were subjected to the elevated plus-maze test. The frequencies of pinning and playful attack were less in the early-weaned group than in the normally weaned group. In the elevated plus-maze test, rat pups in the early-weaned group had higher anxiety levels. The results showed that deprivation of mother-pup interactions during the preweaning period decreases affiliative interactions between cage mates, including play-fighting behaviors during the postweaning developmental period, and increases anxiety levels during early adulthood.     

Maternally separated rats show deficits in maternal care in adulthood

Although there is considerable research on the phenomenology, neuroendocrinology, neuroanatomy, and sensory control of maternal behavior, little is known about the influences of early postnatal and postweaning experiences on the development of maternal behavior. The purpose of this study was to assess how early life separation from the mother rat affects development of the offspring's juvenile and adult maternal behavior. From postnatal Days 1 to 17, 3 female rats within each litter were separated (SEP) from the mother and the rest of the litter for 5 hr daily while 3 of their sisters were not maternally separated (NSEP). On postnatal Day 21, all subjects were weaned and randomly assigned to one of three juvenile conditions. One female from both SEP and NSEP groups was either isolated (I), given a social conspecific (S), or given 1- to 4-day-old pups (P) for 5 consecutive days. Maternal behavior of SEP and NSEP animals was assessed and recorded on each of the 5 days. Once all animals reached adulthood, they were mated, gave birth, and were assessed for their maternal behavior. We found that the effects of maternal separation on juvenile maternal-like behaviors were minimal. On the other hand, maternal separation reduced adult maternal licking and crouching over pups. In addition, there was a significant interaction between postnatal and juvenile experience on maternal crouching in maternal animals. These results are discussed in terms of the variety of possible behavioral, endocrine, and neurochemical mechanisms that mediate the effects of early life experiences on adult maternal behavior.     

大鼠海马γ-氨基丁酸-A受体在生后发育中的表达

目的:研究大鼠生后发育过程中海马组织γ-氨基丁酸-A(GABA-A)受体的表达规律。方法:用免疫组织化学和PCR技术,检测不同年龄大鼠海马组织GABA-A受体及编码该受体的mRNA表达,并用图像分析方法进行定量研究。结果:大鼠海马组织在生后3 d已经出现GABA-A受体免疫反应,以后逐渐增强,到生后30 d达到最高值,海马各区GABA-A受体免疫反应强度没有显著的差别;编码GABA-A受体的mRNA表达也有类似的增龄性增多,但其表达高峰值提前到14 d。结论:生后大鼠海马GABA-A受体表达在一定时期内呈增龄性表达增强的趋势。