Behavior Of Pups Research Articles

Effects of maternal subclinical hypothyroidism on neurodevelopment of offspring- an animal-based study

Background and Objective: Maternal subclinical hypothyroidism (SCH) during pregnancy is associated with adverse maternal and neonatal outcomes, including cognitive and neuropsychiatric effects. This study aimed to assess the impact of maternal SCH on young rats, examining behavioral and gross brain structure changes, and evaluating reversibility after levothyroxine treatment. Methods: This experimental animal study was conducted at the Aga Khan University for a duration of 10 months. The study was approved by the Ethical committee for Animal Care and Use. Fourteen female Sprague Dawley rats were grouped into Treated (induced with SCH, treated with levothyroxine), Untreated (induced with SCH, no treatment), and Control (administered saline). Pups' body weight was monitored, and histological procedures were conducted at postnatal days 7, 14, and 21. Behavioral tests (elevated plus maze, forced swim, and tail suspension) assessed anxiety and depression. The data was analysed using SPSS (version 25.0). The mean, median, and standard deviation were calculated for each quantitative parameter. A one-way ANOVA was performed to identify any variation in mean thyroid levels between groups, and an independent sample t-test was utilized to confirm mean differences in anxiety levels. Results: On postnatal day 7, untreated pups showed lower body weight compared to treated and control groups. This trend continued on days 14 and 21. By day 21, an elevated plus maze (EPM) test indicated anxiety-like behaviour in untreated pups, while they also exhibited more signs of depression in tests like the forced swim test (FST) and tail suspension test (TST). Brain structure, including the prefrontal cortex, remained intact in offspring affected by maternal thyroid dysfunction, with no significant changes observed in brain morphology across the groups. Conclusion: Despite unchanged brain structure, untreated rat pups exhibited significant behavioral differences indicative of depression. This underscores the importance of understanding the behavioral impacts of maternal SCH even in the absence of gross anatomical alterations.

The Role of Play in the Social Development of Grey Seal (Halichoerus grypus) Pups with Comparative Notes on the Harbour Seal (Phoca vitulina).

Juvenile grey seals are known to be highly social, interacting with contact behaviours interpreted as gentle play. However, minimal sociality of pups with their mothers and among weaned pups has been suggested. The present study aimed to observe the natural social interactions of pups to track the early ontogeny of their sociality. Pup behaviour at a salt marsh colony on the east coast of England was video-recorded. Pups interacted with their mothers around suckling bouts and after weaning as they gathered around pools. The records were transcribed to spreadsheets in 30 s time segments to estimate the frequency and co-occurrence of different behaviours. Mother-pup interaction comprised nosing contacts and sometimes contact play, involving one laying the head and fore-flipper over the other. Initial weaned pup encounters involved tentative nosing and defensive splashing, indicating contact shyness. However, socially orientated locomotor play, supine posturing, and exaggerated raising of fore- and hind-flippers led to reduced shyness and pups following one another towards the sea. Archive data on subadult interactions and on harbour seal behaviours were re-analysed. Gentle play-like contact between mother-pup, juvenile, and adult pairs is interpreted here as a universal mode of social bonding, underscoring the social structure of both grey and harbour seals.

Adjusting and validating a procedure for parenteral anaesthesia in neonatal mice.

For neonatal pups, parenteral anaesthesia is said to be not reliable as low doses induce no anaesthesia whereas high doses render high mortality rates. In this work we have adapted parenteral anaesthesia procedures approved for pups >7 days of age, to anaesthetize neonatal animals (postnatal days 3-4; P3-P4) for keeping them immobile for a long period. In our first experiment we analysed the behaviour of P3-P4 mouse pups for 70 min after intraperitoneal administration of low (37.5/3.75 mg/kg) or high (50/5) doses of a ketamine/xylazine anaesthetic mixture, both in the low range as compared with dosages employed in adults. Pups became immobile in ≈7 min and remained immobile for ≈45 min, irrespective of the age and dose of anaesthesia, younger pups (P3) being apparently more sensitive to the dosage. In the second experiment, we studied the response of P3 pups to mildly nociceptive stimulations, performed with a 4.0 g von Frey filament applied to the dorsal aspect of their paws. These stimuli elicited reaction in 100% of the cases in non-anaesthetized pups. The results indicate that the high dose significantly reduced responses as compared with the low dose of anaesthesia. With the low dose, <40% of the pups were unresponsive to nociceptive stimulation, whereas the high dose resulted in 50-60% of the animals not responding. Mortality was low irrespective of age or dose, suggesting that doses can be further increased if needed for invasive experimental procedures.

Effect of icariin on depressive behaviour in rat pups. Evidences for its mechanism of action by integrating network pharmacology, metabolomics and gut microbiota composition

BackgroundPrenatal stress (PS) can cause cognitive disorder and a range of psychological illnesses, including anxiety and depression. Icariin (ICA) has shown promising effects in improving PS-induced depressive behaviour. However, its mechanism of action remains unclear. PurposeThis study was performed to reveal the key targets, metabolites and gut microbiota for ICA in improving depressive behaviour in PS rat pups. MethodsA prenatal restraint stress animal model was established for Sprague–Dawley (SD) rats in late pregnancy. Male pups were randomly divided into six groups: no stress group (NS), PS group, PS + saline group (PS_S), PS + high-dose ICA group (ICAH, 80 mg/kg*day), PS + low-dose ICA group (ICAL, 40 mg/kg*day) and PS + fluoxetine group (FLU, 10 mg/kg*day). The depressive behaviour of each group of rat pups was evaluated using open field test, forced swimming test and sucrose preference test. Different metabolites were identified using untargeted metabolomics of serum and faeces, and metabolic pathways were analyzed through MetaboAnalyst. Targets for ICA acting on depression were determined after network pharmacology was applied. An integrated network of network pharmacology and metabolomics were constructed using Cytoscape software, and molecular docking were performed to verify the interactions between ICA and key targets. Finally, gut microbiota of rat pups in each group were analyzed after 16S rDNA sequencing. ResultsPS could cause rat pups to exhibit depressive behaviour, and ICA could significantly improve this depressive behaviour. A total of 49 differential metabolites were found in serum and 23 differential metabolites were found in faeces, and 24 metabolites in serum and 6 metabolites in faeces could be reversed following ICA administration. Integrated analysis focused on five key targets (i.e. adenosyl homocysteinase; medium-chain specific acyl-CoA dehydrogenase, mitochondrial; thymidine phosphorylase; cGMP-specific 3′,5′-cyclic phosphodiesterase and xanthine dehydrogenase/oxidase) and three metabolites (i.e. palmitoylcarnitine, methionine and hypoxanthine). Molecular docking indicated that ICA combined well with key targets. Gut microbiota analysis showed that g_Bacteroides, f_Bacteroidaceae and s_Lactobacillus reuteri were required for ICA to improve depressive behaviour. ConclusionIn this study, the antidepressant mechanism of ICA was clarified with a strategy of integrating metabolomics, network pharmacology and gut microbiota. ICA has a good effect on improving metabolism and increasing the abundance of probiotics in the intestine. The present research provided new insights into the anti-depressant mechanism of ICA.

2-Chloro-3,7,8-tribromodibenzofuran as a new environmental pollutant inducing atypical ultrasonic vocalization in infant mice.

Epidemiological and experimental studies indicate that maternal exposure to environmental pollutants impairs the cognitive and motor functions of offspring in humans and laboratory animals. Infant ultrasonic vocalizations (USVs), the communicative behavior of pups toward caregivers, are impaired in rodent models of neurodevelopmental disorders, suggesting a useful method to evaluate the developmental neurotoxicity of environmental pollutants. Therefore, we investigated USVs emitted by mouse pups of dams exposed to 2-chloro-3,7,8-tribromodibenzofuran (TeXDF) and 1,2,3,7,8-pentabromodibenzofuran (PeBDF), which are detected in the actual environment. The USV duration and number in the pups born to dams administered with TeXDF 40μg/kg body weight (b.w.), but not 8μg/kg b.w., on gestational day (GD) 12.5, were significantly lower than those in the corresponding pups on postnatal days 3-9. Conversely, there was no statistical change in the USVs emitted by the pups of dams administered with PeBDF 35 or 175μg/kg b.w. on GD 12.5. To examine whether maternal exposure leads to behavioral impairments in adulthood, we analyzed exploratory behaviors in a novel environment using IntelliCage, a fully automated testing apparatus for group-housed mice. Neither TeXDF nor PeBDF exposure induced significant differences in offspring exploration. Considered together, our findings revealed that TeXDF induces atypical USV emission in infant mice, suggesting the importance of further studies on the risk assessment of mixed brominated/chlorinated dibenzo-p-dioxins and dibenzofurans.

Gestational paracetamol exposure induces core behaviors of neurodevelopmental disorders in infant rats and modifies response to a cannabinoid agonist in females

Paracetamol (PAR) is an over-the-counter analgesic/antipyretic used during pregnancy worldwide. Epidemiological studies have been associating gestational PAR exposure with neurobehavioral alterations in the progeny resembling autism spectrum disorders and attention-deficit hyperactivity disorder symptoms. The endocannabinoid (eCB) dysfunction was previously hypothesized as one of the modes of action by which PAR may harm the developing nervous system. We aimed to evaluate possible effects of gestational exposure to PAR on male and female rat's offspring behavior and if an acute injection of WIN 55,212–2 (WIN, 0.3 mg/kg), a non-specific cannabinoid agonist, prior to behavioral tests, would induce different effects in PAR exposed and non-exposed animals. Pregnant Wistar rats were gavaged with PAR (350 mg/kg/day) or water from gestational day 6 until delivery. Nest-seeking, open field, apomorphine-induced stereotypy, marble burying and three-chamber tests were conducted in 10-, 24-, 25- or 30-days-old rats, respectively. PAR exposure resulted in increased apomorphine-induced stereotyped behavior and time spent in the central area of the open field in exposed female pups. Additionally, it induced hyperactivity in the open field and increased marble burying behavior in both male and female pups. WIN injection modified the behavioral response only in the nest seeking test, and opposite effects were observed in control and PAR-exposed neonate females. Reported alterations are relevant for the neurodevelopmental disorders that have been associated with maternal PAR exposure and suggest that eCB dysfunction may play a role in the action by which PAR may harm the developing brain.

Postnatal melatonin administration to stressed dams for ameliorating risk-taking behaviour in rat pups through maternal care improvement.

Anxiety often occurs both concurrently and sequentially in childhood and adolescence in association with prenatal stress, which may reduce the quality of maternal care and then cause mood disorders among children in later life. Against this background, melatonin, as a powerful antioxidant, was used in the present study to ameliorate risk-taking behaviour induced by pure maternal care in rat pups. The Wistar rat dams recruited in this study were exposed to restraint stress from gestational day (GD) 11 until delivery. They further received melatonin (10mg/kg) during the postnatal days (PNDs) 0-7 by intraperitoneal (IP) injections at 4:00 PM. The pregnant rats were then divided into four groups, namely, control, stress, stress + melatonin and melatonin, and their maternal behaviour and corticosterone levels were measured. In the offspring, the outcomes of some behavioural tasks, including the elevated plus-maze (EPM) and open-field (OF) tests were ultimately assessed. The study results revealed that the quantity and quality of maternal care significantly declined and the plasma corticosterone levels compounded in the stressed dams. Melatonin treatment, however, improved their nursing behaviour and reduced their plasma corticosterone levels. The offspring performance in two tasks also showed an upward trend in risk-taking behaviour in the stress group, and melatonin administration ameliorated the effects of stress and lessened their anxiety-like behaviour. It was concluded that prenatal restraint stress could impair stress responses and quality of maternal care, whereas postnatal melatonin administration potentially contributed to the normalization of stress reaction and anxiolysis.

Effect of troxerutin consumption during gestation period on reflexive motor behavior in mice offspring.

This study aimed to determine the effects of troxerutin consumption during gestation on reflexive motor behavior in mice offspring. Forty pregnant female mice were allocated into four groups. In the control group, mice received water, while in groups 2-4, female mice p.o. administered troxerutin (50, 100, and 150 mg/kg) at 5, 8, 11, 14, and 17 days of gestation (GD). Following delivery, pups were selected based on their experimental group, and reflexive motor behaviors were determined. Also, serum malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPx), and total antioxidant status (TAS) were determined. Based on the findings, maternal exposure to troxerutin (100 and 150 mg/kg) increased ambulation scores in offspring's compared with control group (P <0.05). Also, prenatal exposure to troxerutin increased front- and hind-limb suspension scores in newborns compared with control group (P <0.05). Maternal exposure to troxerutin increased grip strength and negative geotaxis in newborns in comparison with control mice (P <0.05). Prenatal exposure to troxerutin (100 and 150 mg/kg) decreased hind-limb foot angle and surface righting in pups compared with control group (P <0.05). Maternal exposure to troxerutin decreased MDA production and increased SOD, GPx, and TAS levels in offspring (P <0.05). These results suggested that prenatal consumption of the troxerutin improves reflexive motor behaviors in mice pups.

The influence of maternal separation on the development of voiding and behavior in rat pups

Aim:This study aimed to investigate the effects of early-life stress resulting from maternal separation (MS) on urinary function and behavioral changes. Methods:We categorized 85 rat pups into control and MS groups. Two groups of each sex during post-weaning (3 weeks old) and maturity (9 weeks old) were compared. Urinary function was evaluated using metabolic cages and continuous cystometry. Morphological changes were assessed based on bladder weight and staining. In behavioral experiments, spatial working memory using the Y-maze and anxiety-like behavior in the elevated plus-maze were assessed. Results:In the metabolic cage test, the single voiding volume decreased in the 3-week-old male and female MS groups. In continuous cystometry, the intercontraction interval was shortened in the 9-week-old male and female MS groups. MS showed a significant change in bladder weight and wall thickness in 3-week-old females. Conversely, a change in bladder wall thickening was observed in male rats aged 9 weeks. In the Y-maze, no differences were found in alternation. In the elevated plus-maze test, the 3-week-old MS groups of both sexes had fewer entries into the open arm and slower mean velocities, while the 9-week-old groups had a greater number of entries into the open arm and a faster average speed. Conclusion:MS affects voiding behavior and bladder activity during the early phase of maturation. After growth, bladder overactivity and less anxiety-like hyperactivity co-exist, suggesting that early-life stress due to MS may have lasting effects on common neural circuits related to urinary function and behavior.

Rates of ultrasonic vocalizations are more strongly related than acoustic features to non-vocal behaviors in mouse pups.

Mouse pups produce. ultrasonic vocalizations (USVs) in response to isolation from the nest (i.e., isolation USVs). Rates and acoustic features of isolation USVs change dramatically over the first two weeks of life, and there is also substantial variability in the rates and acoustic features of isolation USVs at a given postnatal age. The factors that contribute to within age variability in isolation USVs remain largely unknown. Here, we explore the extent to which non-vocal behaviors of mouse pups relate to the within age variability in rates and acoustic features of their USVs. We recorded non-vocal behaviors of isolated C57BL/6J mouse pups at four postnatal ages (postnatal days 5, 10, 15, and 20), measured rates of isolation USV production, and applied a combination of pre-defined acoustic feature measurements and an unsupervised machine learning-based vocal analysis method to examine USV acoustic features. When we considered different categories of non-vocal behavior, our analyses revealed that mice in all postnatal age groups produce higher rates of isolation USVs during active non-vocal behaviors than when lying still. Moreover, rates of isolation USVs are correlated with the intensity (i.e., magnitude) of non-vocal body and limb movements within a given trial. In contrast, USVs produced during different categories of non-vocal behaviors and during different intensities of non-vocal movement do not differ substantially in their acoustic features. Our findings suggest that levels of behavioral arousal contribute to within age variability in rates, but not acoustic features, of mouse isolation USVs.

Supplementation of L-Ornithine Could Increase Sleep-like Behavior in the Mouse Pups.

Along the maternal-fetal-neonatal axis, one of the problems relating to the maternal-neonatal axis is infant sleep problems including nighttime crying. One possible solution could be to provide the newborn with sleep-promoting ingredients through breast milk or formula. So far, it has been reported that L-ornithine has a sleep-related effect. Therefore, we investigated the effect of dietary L-ornithine on maternal mouse plasma and milk L-ornithine levels in Experiment 1. In Experiment 2, a single dose of L-ornithine was applied to know the time-course changes in plasma, mammary gland and milk L-ornithine levels. Experiment 3 was conducted to confirm sleep behavior as well as changes in polyamine levels in milk. L-Ornithine levels in maternal plasma significantly increased by both dietary regimen and single oral administration in Experiments 1 and 2. Both L-ornithine treatments also increased its levels in milk, although not to a concentration as high as in plasma. In Experiment 3, the level of polyamines, which are metabolized from L-ornithine, did not significantly differ after L-ornithine administration. In sleep-like behavior observations, the average concentration of L-ornithine in milk did not increase the sleep-like behavior of mouse pups. However, more concentrated L-ornithine solutions can significantly increase sleep-like behavior. These results revealed that even if mothers ingested L-ornithine to increase L-ornithine levels in breast milk, it is difficult to promote sleep in newborns. Because it is difficult to raise L-ornithine in breast milk to sleep-inducing levels, L-ornithine added formula may partially improve infant sleep and has the potential for preventing infant sleep problems such as nighttime crying.

Prolactin Action Is Necessary for Parental Behavior in Male Mice.

Parental care is critical for successful reproduction in mammals. Recent work has implicated the hormone prolactin in regulating male parental behavior, similar to its established role in females. Male laboratory mice show a mating-induced suppression of infanticide (normally observed in virgins) and onset of paternal behavior 2 weeks after mating. Using this model, we sought to investigate how prolactin acts in the forebrain to regulate paternal behavior. First, using c-fos immunoreactivity in prolactin receptor (Prlr) Prlr-IRES-Cre-tdtomato reporter mouse sires, we show that the circuitry activated during paternal interactions contains prolactin-responsive neurons in multiple sites, including the medial preoptic nucleus, bed nucleus of the stria terminalis, and medial amygdala. Next, we deleted Prlr from three prominent cell types found in these regions: glutamatergic, GABAergic, and CaMKIIα. Prlr deletion from CaMKIIα, but not glutamatergic or GABAergic cells, had a profound effect on paternal behavior as none of these KO males completed the pup-retrieval task. Prolactin was increased during mating, but not in response to pups, suggesting that the mating-induced secretion of prolactin is important for establishing the switch from infanticidal to paternal behavior. Pharmacological blockade of prolactin secretion at mating, however, had no effect on paternal behavior. In contrast, suppressing prolactin secretion at the time of pup exposure resulted in failure to retrieve pups, with exogenous prolactin administration rescuing this behavior. Together, our data show that paternal behavior in sires is dependent on basal levels of circulating prolactin acting at the time of interaction with pups, mediated through Prlr on CaMKIIα-expressing neurons.SIGNIFICANCE STATEMENT Parental care is critical for offspring survival. Compared with maternal care, however, the neurobiology of paternal care is less well understood. Here we show that the hormone prolactin, which is most well known for its female-specific role in lactation, has a role in the male brain to promote paternal behavior. In the absence of prolactin signaling specifically during interactions with pups, father mice fail to show normal retrieval behavior of pups. These data demonstrate that prolactin has a similar action in both males and females to promote parental care.

Maternal lead exposure induces sex-dependent cerebellar glial alterations and repetitive behaviors

Lead (Pb) is one of the most prevalent heavy metals we encounter daily. Although there are many reports regarding their toxic effects on humans, the effects of exposure to low lead concentrations throughout the pregnancy period on the offspring are not fully elucidated yet. This study aimed to investigate the cellular mechanisms that occur in response to lead exposure. To this end, we administered lead-containing water to pregnant mice from the day of conception till delivery or till day 28 postnatally. Furthermore, we performed neurodevelopmental disorder-related behavior tests and RNA-sequencing analysis. We used both genders for all experiments because neurodevelopmental disorders usually show several sex-dependent differences. The results revealed increased levels of gliosis in the cerebella of lead-exposed pups compared to those in littermates belonging to the control group. Additionally, we observed altered behaviors of male mice in the autism spectrum disorder-related tests. RNA-sequencing results revealed changes in gamma-aminobutyric acid (GABA) signaling in the lead-exposed mouse model. Specifically, the lead-exposed male mice showed decreased monoamine oxidase B and increased levels of diamine oxidase enzyme, which is related to the synthesis of GABA in astrocytes. These findings demonstrate sex-dependent basal developmental changes in glial cells and an increased prevalence of autistic-like behaviors in the young pups of mothers exposed to lead during pregnancy.

Newly Documented Behaviour of Free-Ranging Arctic Wolf Pups

Whereas much is known about the behavior and development of captive young wolf (Canis lupus) pups, less detail has been published about some aspects of free-ranging wolf pup behavior. This article synthesizes 42 observations of free-ranging Arctic wolf pups from ages 13 through 52 days made during 10 summers from 1987 through 2006 on Ellesmere Island, Nunavut, Canada. Besides listing key behaviors such as howling and caching, I record unique observations of ages of pup urination without adult stimulation (22, 33, 42, 52 days), knowledge of which is important to studies of wolf domestication, and of a 48-day-old pup that traveled 39 km during a 12 h and 19 min round trip between the den and a prey carcass, including a 26.5 km trek in 5 h. These observations should lead to a deeper and more complete understanding of this critical period of pup growth and development.

Some features of the brain of Wistar rats that developed in broods of various numbers

Objective: To study the morphofunctional features of the brain development of 14- and 30-day-old rats born in broods that differ significantly in numbers.Methods: 2 age groups of animals were studies – 14-day-old and 30-day-old rat pups, the offspring of intact rats, each group was divided into subgroups: 1) small broods (4-7 rat pups per brood) and 2) broods of average size (9-12 rats in a brood). The brains of animals of both subgroups were subjected to morphometric and histochemical studies. At the age of 28 days, rat pups from the second age group were tested in an elevated plus maze.Results: It was found that rat pups from small broods at the age of 14 and 30 days had signs of acceleration and differed in the outstripping rates of development of brain from animals from the group of broods of average size. Differences were manifested by a greater mass of the brain and hemisphere, a lower numerical density of neurons and a higher density of gliocytes, a large glioneural index, as well as the size of neurons, their nuclei and nucleoli in the cortex of the parietal lobe and the hippocampus. The behavior of pups from small broods and broods of average numbers at the age of 28 days in an elevated plus maze had significant intergroup differences.Conclusions: These findings indicate that differences in brood numbers affect important indicators of brain development.

Moderate Folic Acid Supplementation in Pregnant Mice Results in Altered Sex-Specific Gene Expression in Brain of Young Mice and Embryos.

Food fortification and increased vitamin intake have led to higher folic acid (FA) consumption by many pregnant women. We showed that FA-supplemented diet in pregnant mice (fivefold higher FA than the recommended level (5xFASD)) led to hyperactivity-like behavior and memory impairment in pups. Disturbed choline/methyl metabolism and altered placental gene expression were identified. The aim of this study was to examine the impact of 5xFASD on the brain at two developmental stages, postnatal day (P) 30 and embryonic day (E) 17.5. Female C57BL/6 mice were fed a control diet or 5xFASD for 1 month before mating. Diets were maintained throughout the pregnancy and lactation until P30 or during pregnancy until E17.5. The 5xFASD led to sex-specific transcription changes in P30 cerebral cortex and E17.5 cerebrum, with microarrays showing a total of 1003 and 623 changes, respectively. Enhanced mRNA degradation was observed in E17.5 cerebrum. Expression changes of genes involved in neurotransmission, neuronal growth and development, and angiogenesis were verified by qRT-PCR; 12 and 15 genes were verified at P30 and E17.5, respectively. Hippocampal collagen staining suggested decreased vessel density in FASD male embryos. This study provides insight into the mechanisms of neurobehavioral alterations and highlights potential deleterious consequences of moderate folate oversupplementation during pregnancy.

Are there long-term effects of early neurological stimulation (ENS) on working dogs?

Early neurological stimulation (ENS) has been defined as the application of five specific brief daily manipulations to pups from birth until 2–3 weeks of age (Battaglia, 2009). This approach has been adopted by many kennels and promoted as a means to improve the future performance of working dogs. Although there is ample evidence that enrichment and socialization have a positive impact on adult behavior in dogs, there is no evidence ENS in and of itself has any lasting effect on dog behavior. Since the purported benefits are large and the required manipulations are minor, we sought: a) to evaluate the effects of ENS on self-confidence, motivation, and aggressive/defensive behavior in pups from 2 months of age until 12 months; and b) to determine whether ENS increased the probability of dogs being considered as suitable for further training as working dogs. We used a split-litter design where half of the pups received the ENS manipulations, and the other half were a control group that was simply held for the same amount of time required by the ENS manipulations. Our results indeed show that the ENS treated dogs are more likely to be accepted for such training than were the control pups but that differences in behavior only appeared at testing on months 10 and 12. We suggest two reasons for our results. First, immediately after the ENS manipulations, the ENS pups probably were a bit more socialized than the control pups. This resulted in the caregivers spending more time with the ENS pups which further increased the discrepancy in socialization between the ENS and control pups. Second, since the caregivers were aware of the ENS manipulation their expectations resulted in more positive interactions with the pups resulting in improved behavior; a phenomenon known as the Pygmalion effect.

Deficiency of the paternally inherited gene Magel2 alters the development of separation-induced vocalization and maternal behavior in mice.

The behavior of offspring results from the combined expression of maternal and paternal genes. Genomic imprinting silences some genes in a parent-of-origin specific manner, a process that, among all animals, occurs only in mammals. How genomic imprinting affects the behavior of mammalian offspring, however, remains poorly understood. Here, we studied how the loss of the paternally inherited gene Magel2 in mouse pups affects the emission of separation-induced ultrasonic vocalizations (USV). Using quantitative analysis of more than 1000 USVs, we characterized the rate of vocalizations as well as their spectral features from postnatal days 6-12 (P6-P12), a critical phase of mouse development that covers the peak of vocal behavior in pups. Our analyses show that Magel2 deficient offspring emit separation-induced vocalizations at lower rates and with altered spectral features mainly at P8. We also show that dams display altered behavior towards their own Magel2 deficient offspring at this age. In a test to compare the retrieval of two pups, dams retrieve wildtype control pups first and faster than Magel2 deficient offspring. These results suggest that the loss of Magel2 impairs the expression of separation-induced vocalization in pups as well as maternal behavior at a specific age of postnatal development, both of which support the pups' growth and development.

Evaluation of the ultradiluted medication MPD 30 CH in the offspring of mice mothers treated with methylphenidate during lactation.

Background: Methylphenidate (MPD) is a non-stimulating amphetamine that has being used for some time in the treatment of Attention Deficit Hyperactivity Disorder (ADHD) and, in adequate doses, it promotes the remission of symptoms and the improvement of important aspects, as social interaction and academic performance1, in patients with ADHD. Literature data indicates that MPD attenuates maternal behavior in mices2. According to this line of study, the work “Repeated methylphenidate administration during lactation reduces maternal behavior, induces maternal tolerance, and increases anxiety-like behavior in pups in adulthood”3 was carried out and confirmed that MPD administration during early lactation disrupts maternal behavior and causes anxiety in pups in adulthood. Would it be possible that ultradiluted and dynamized MPD change pups’ behavior? Objective: The aim of this study was to evaluate how the ultradiluted drug may or may not change the behavior of the animals at issue.&#x0D; Material and Methods: The medication was prepared according to the Brazilian Homeopathic Pharmacopeia, in the 30 CH dilution. The present study was approved by the Ethics Committee for Animal Experimentation of the Paulista University (No. 256/14 CEP / ICS / UNIP). Animals in this study were the same of the study above mentioned, and already published. Adult male mice were grouped among 13 animals of the experimental group (adults, offspring of mothers that received MPD during pregnancy) and 9 animals from the mother-control group, which did not take MPD during pregnancy. The 22 animals took ultradiluted MPD 30 CH medication in their drinking water ad libitum, for 20 days. In each water drinker, 5 drops of medication were added and stirred. Behavioral tests, such as the Open Field and the Light Dark Transition Test for mice, were performed. Data was analyzed statistically by the Student's T-Test to compare parametric data from two groups and the Mann-Whitney Test for nonparametric data, where p ≤ 0.05 is considered significant. &#x0D; Results and Discussion: In the Open Field Test, from the group of mothers medicated with methylphenidate during pregnancy, before the medication MPD 30 CH, animals showed a lower mobility and a greater immobility (p≤ 0.05) compared to the control-animals; after medication with the MPD 30 CH, animals exhibited an increase in mobility and a decrease in immobility, leading to no statistical difference between the medication group and the control group. In the Light Dark Transition Test for mice, experimental animals spent more time in the dark box and exhibited a decrease of the Rearing, presenting an anxious behavior; after the MPD medication, there were no more differences between experimental and control groups. Indiscriminate use of amphetamines has increased in recent years and this use, when not monitored, can cause serious adverse effects4. In this sense, ultradiluted medication can collaborate with the remission of possible undesirable effects. Conclusion: Initially, the MPD 30 CH changed the behavior shown by animals born from mothers that took methylphenidate during lactation and presented an anxious behavior as an unexpected effect. The mice that took the MPD 30 CH did not present the anxious behavior. Other experiments should be conducted to confirm the results of this study.

A Neural Network Reveals Motoric Effects of Maternal Preconception Exposure to Nicotine on Rat Pup Behavior: A New Approach for Movement Disorders Diagnosis.

Neurodevelopmental disorders can stem from pharmacological, genetic, or environmental causes and early diagnosis is often a key to successful treatment. To improve early detection of neurological motor impairments, we developed a deep neural network for data-driven analyses. The network was applied to study the effect of maternal nicotine exposure prior to conception on 10-day-old rat pup motor behavior in an open field task. Female Long-Evans rats were administered nicotine (15 mg/L) in sweetened drinking water (1% sucralose) for seven consecutive weeks immediately prior to mating. The neural network outperformed human expert designed animal locomotion measures in distinguishing rat pups born to nicotine exposed dams vs. control dams (87 vs. 64% classification accuracy). Notably, the network discovered novel movement alterations in posture, movement initiation and a stereotypy in “warm-up” behavior (repeated movements along specific body dimensions) that were predictive of nicotine exposure. The results suggest novel findings that maternal preconception nicotine exposure delays and alters offspring motor development. Similar behavioral symptoms are associated with drug-related causes of disorders such as autism spectrum disorder and attention-deficit/hyperactivity disorder in human children. Thus, the identification of motor impairments in at-risk offspring here shows how neuronal networks can guide the development of more accurate behavioral tests to earlier diagnose symptoms of neurodevelopmental disorders in infants and children.