Depression during pregnancy is detrimental for the wellbeing of the expectant mother and can exert long-term consequences on the offspring\'s development and mental health. In this context, both the gestational environment and the postpartum milieu may be negatively affected by the depressive pathology. It is, however, challenging to assess whether the contributions of prenatal and postnatal depression exposure are distinct, interactive, or cumulative, as it is unclear whether antenatal effects are due to direct effects on fetal development or because antenatal symptoms continue postnatally. Preclinical models have sought to answer this question by implementing stressors that induce a depressive-like state in the dams during pregnancy and studying the effects on the offspring. The aim of our present study was to disentangle the contribution of direct stress in utero from possible changes in maternal behavior in a novel model of preconceptional stress based on social isolation rearing (SIR). Using a cross-fostering paradigm in this model, we show that while SIR leads to subtle changes in maternal behavior, the behavioral changes observed in the offspring are driven by a complex interaction between sex, and prenatal and postnatal maternal factors. Indeed, male offspring are more sensitive to the prenatal environment, as demonstrated by behavioral and transcriptional changes driven by their birth mother, while females are likely affected by more complex interactions between the pre and the postpartum milieu, as suggested by the important impact of their surrogate foster mother. Taken together, our findings suggest that male and female offspring have different time-windows and behavioral domains of susceptibility to maternal preconceptional stress, and thus underscore the importance of including both sexes when investigating the mechanisms that mediate the negative consequences of exposure to such stressor.

Selective breeding has been utilized to study the genetic basis of exercise behavior, but research suggests that epigenetic mechanisms, such as DNA methylation, also contribute to this behavior. In a previous study, we demonstrated that the brains of mice from a genetically selected high runner (HR) line have sex-specific changes in DNA methylation patterns in genes known to be genomically imprinted compared to those from a non-selected control (C) line. Through cross-fostering, we also found that maternal upbringing can modify the DNA methylation patterns of additional genes. Here, we identify an additional set of genes in which DNA methylation patterns and gene expression may be altered by selection for increased wheel-running activity and maternal upbringing. We performed bisulfite sequencing and gene expression assays of 14 genes in the brain and found alterations in DNA methylation and gene expression for Bdnf, Pde4d and Grin2b. Decreases in Bdnf methylation correlated with significant increases in Bdnf gene expression in the hippocampus of HR compared to C mice. Cross-fostering also influenced the DNA methylation patterns for Pde4d in the cortex and Grin2b in the hippocampus, with associated changes in gene expression. We also found that the DNA methylation patterns for Atrx and Oxtr in the cortex and Atrx and Bdnf in the hippocampus were further modified by sex. Together with our previous study, these results suggest that DNA methylation and the resulting change in gene expression may interact with early-life influences to shape adult exercise behavior.

At birth, mammals experience a massive colonization by microorganisms. We previously reported that newborn mice gestated and born germ-free (GF) have increased microglial labeling and alterations in developmental neuronal cell death in the hippocampus and hypothalamus, as well as greater forebrain volume and body weight when compared to conventionally colonized (CC) mice. To test whether these effects are solely due to differences in postnatal microbial exposure, or instead may be programmed in utero, we cross-fostered GF newborns immediately after birth to CC dams (GF→CC) and compared them to offspring fostered within the same microbiota status (CC→CC, GF→GF). Because key developmental events (including microglial colonization and neuronal cell death) shape the brain during the first postnatal week, we collected brains on postnatal day (P) 7. To track gut bacterial colonization, colonic content was also collected and subjected to 16S rRNA qPCR and Illumina sequencing. In the brains of GF→GF mice, we replicated most of the effects seen previously in GF mice. Interestingly, the GF brain phenotype persisted in GF→CC offspring for almost all measures. In contrast, total bacterial load did not differ between the CC→CC and GF→CC groups on P7, and bacterial community composition was also very similar, with a few exceptions. Thus, GF→CC offspring had altered brain development during at least the first 7 days after birth despite a largely normal microbiota. This suggests that prenatal influences of gestating in an altered microbial environment programs neonatal brain development.

As environmental fluctuations are becoming more common, organisms need to rapidly adapt to anthropogenic, climatic, and ecological changes. Epigenetic modifications and DNA methylation in particular provide organisms with a mechanism to shape their phenotypic responses during development. Studies suggest that environmentally induced DNA methylation might allow for adaptive phenotypic plasticity that could last throughout an organism\'s lifetime. Despite a number of studies demonstrating environmentally induced DNA methylation changes, we know relatively little about what proportion of the epigenome is affected by environmental factors, rather than being a consequence of genetic variation. In the current study, we use a partial cross-foster design in a natural great tit (Parus major) population to disentangle the effects of common origin from common rearing environment on DNA methylation. We found that variance in DNA methylation in 8,315 CpG sites was explained by a common origin and only in 101 by a common rearing environment. Subsequently, we mapped quantitative trait loci for the brood of origin CpG sites and detected 754 cis and 4,202 trans methylation quantitative trait loci, involving 24% of the CpG sites. Our results indicate that the scope for environmentally induced methylation marks independent of the genotype is limited and that the majority of variation in DNA methylation early in life is determined by genetic factors instead. These findings suggest that there may be little opportunity for selection to act on variation in DNA methylation. This implies that most DNA methylation variation likely does not evolve independently of genomic changes.

Many social animals display collective activity cycles based on synchronous behavioural oscillations across group members. A classic example is the colony cycle of army ants, where thousands of individuals undergo stereotypical biphasic behavioural cycles of about one month. Cycle phases coincide with brood developmental stages, but the regulation of this cycle is otherwise poorly understood. Here, we probe the regulation of cycle duration through interactions between brood and workers in an experimentally amenable army ant relative, the clonal raider ant. We first establish that cycle length varies across clonal lineages using long-term monitoring data. We then investigate the putative sources and impacts of this variation in a cross-fostering experiment with four lineages combining developmental, morphological and automated behavioural tracking analyses. We show that cycle length variation stems from variation in the duration of the larval developmental stage, and that this stage can be prolonged not only by the clonal lineage of brood (direct genetic effects), but also of the workers (indirect genetic effects). We find similar indirect effects of worker line on brood adult size and, conversely (but more surprisingly), indirect genetic effects of the brood on worker behaviour (walking speed and time spent in the nest).

We have previously shown that high runner (HR) mice (from a line genetically selected for increased wheel-running behavior) have distinct, genetically based, neurobiological phenotypes as compared with non-selected control (C) mice. However, developmental programming effects during early life, including maternal care and parent-of-origin-dependent expression of imprinted genes, can also contribute to variation in physical activity. Here, we used cross-fostering to address two questions. First, do HR mice have altered DNA methylation profiles of imprinted genes in the brain compared to C mice? Second, does maternal upbringing further modify the DNA methylation status of these imprinted genes? To address these questions, we cross-fostered all offspring at birth to create four experimental groups: C pups to other C dams, HR pups to other HR dams, C pups to HR dams, and HR pups to C dams. Bisulfite sequencing of 16 imprinted genes in the cortex and hippocampus revealed that the HR line had altered DNA methylation patterns of the paternally imprinted genes, Rasgrf1 and Zdbf2, as compared with the C line. Both fostering between the HR and C lines and sex modified the DNA methylation profiles for the paternally expressed genes Mest, Peg3, Igf2, Snrpn, and Impact. Ig-DMR, a gene with multiple paternal and maternal imprinted clusters, was also affected by maternal upbringing and sex. Our results suggest that differential methylation patterns of imprinted genes in the brain could contribute to evolutionary increases in wheel-running behavior and are also dependent on maternal upbringing and sex.

Cross-fostering is a practice commonly used in the swine industry to equalize litter sizes, however, there is limited understanding of the optimum cross-fostering methods that will maximize piglet preweaning growth and survival. This study evaluated the effects of within-litter variation in birth weight after cross-fostering on piglet preweaning mortality (PWM) and weaning weight (WW) using litters of 15 piglets. A hierarchical incomplete block design was used (blocking factors: day of farrowing and sow parity, body condition score, and number of functional teats) with a 3 × 2 factorial arrangement of treatments: 1) Birth Weight Category (BWC): Light (<1.0 kg), Medium (1.0 to 1.5 kg), or Heavy (1.5 to 2.0 kg); 2) Litter Composition: UNIFORM (all 15 piglets in each litter of the same BWC), or MIXED (five piglets in each litter from each BWC, i.e., five Light, five Medium, and five Heavy piglets). At 24 h after birth, piglets were weighed and randomly allotted to litter composition treatments from within BWC. The experimental unit was five piglets of the same BWC; there were three experimental units within each Litter Composition treatment litter. There were 17 blocks, each of six litters (one UNIFORM litter of each BWC; three MIXED litters) and 51 replicates (three replicates per block of six litters) for a total of 102 cross-fostered litters and 1,530 piglets. Piglets were weaned at 19.7 ± 0.46 d of age; WW and PWM were measured. PROC GLIMMIX and MIXED of SAS were used to analyze PWM and WW, respectively. Models included BWC, Litter Composition, the interaction, and replicate within the block. There were BWC by Litter Composition treatment interactions (P ≤ 0.05) for PWM and WW. Preweaning mortality was greater (P ≤ 0.05) for Light piglets in MIXED than UNIFORM litters. In contrast, for Heavy piglets, PWM was greater (P ≤ 0.05) and WW was lower (P ≤ 0.05) in UNIFORM than MIXED litters. Medium piglets had similar (P > 0.05) PWM and WW in UNIFORM and MIXED litters. The results of this study, which involved large litter sizes typical of current commercial production, suggested that for piglet survival to weaning, using cross-fostering to form litters of piglets of similar birth weight was beneficial for light piglets, detrimental for heavy piglets, and neutral for medium piglets.

The Developmental Origins of Health and Disease (DOHaD) concept has been proposed to explain the influence of environmental conditions during critical developmental stages on the risk of diseases in adulthood. The aim of this study was to compare the impact of the prenatal vs. postnatal environment on the gut microbiota in dams during the preconception, gestation and lactation periods and their consequences on metabolic outcomes in offspring. Here we used the cross-fostering technique, e.g. the exchange of pups following birth to a foster dam, to decipher the metabolic effects of the intrauterine versus postnatal environmental exposures to a polyphenol-rich cranberry extract (CE). CE administration to high-fat high-sucrose (HFHS)-fed dams improved glucose homeostasis and reduced liver steatosis in association with a shift in the maternal gut microbiota composition. Unexpectedly, we observed that the postnatal environment contributed to metabolic outcomes in female offspring, as revealed by adverse effects on adiposity and glucose metabolism, while no effect was observed in male offspring. In addition to the strong sexual dimorphism, we found a significant influence of the nursing mother on the community structure of the gut microbiota based on α-diversity and β-diversity indices in offspring. Gut microbiota transplantation (GMT) experiments partly reproduced the observed phenotype in female offspring. Our data support the concept that the postnatal environment represents a critical window to influence future sex-dependent metabolic outcomes in offspring that are causally but partly linked with gut microbiome alterations.

Litter sizes of commercial sows have increased considerably over recent decades, and often exceed the number of functional teats on the sow. The objective of this study was to evaluate the effect of litter size after cross-fostering relative to sow functional teat number on piglet preweaning growth and mortality. A total of 39 litters (561 piglets) were used in a randomized complete block design; blocking factors were farrowing day and sow parity, body condition score, and functional teat number. Three Litter Size treatments were compared (relative to sow functional teat number): Decreased (two piglets less); Control (same number of piglets); Increased (two piglets more). Piglets were randomly allotted to treatment at 24 h after birth to form litters of the appropriate size, with similar mean and CV of birth weight within block. Weaning weights (WW) were collected at 19.5 ± 0.50 d of age; preweaning mortality (PWM) was recorded. Litter sizes were between 11 and 17 piglets, depending on block and treatment. The Decreased treatment had lower (P ≤ 0.05) PWM than the Increased (7.7% and 17.9%, respectively); the Control was intermediate (11.5%) and not different (P > 0.05) from the other treatments. The rate of decline in litter size from birth to weaning was greater (P ≤ 0.05) for the Increased than the Decreased treatment (-0.16 vs. -0.05 piglets per day), with the Control (-0.09 piglets per day) being intermediate and different (P ≤ 0.05) to the other two treatments. Litter sizes at weaning were greater (P ≤ 0.05) for the Increased than the Decreased treatment (13.3 and 11.3, respectively); the Control treatment was intermediate (12.6) and not different (P > 0.05) to the other treatments. The log odds of PWM increased with the decreasing birth weight, at a similar rate (P > 0.05) for all Litter Size treatments. However, the intercept was greater (P ≤ 0.05) for the Increased compared with the Decreased treatment; the Control was intermediate and different (P > 0.05) to the other two treatments. Mean WW tended (P = 0.07) to be greater for the Decreased (6.17 kg) compared to the Control and Increased treatments (5.86 and 5.84 kg, respectively). In conclusion, increasing litter size after cross-fostering relative to the number of functional teats of the sow increased piglet PWM, and tended to decrease WW.

Cross-fostering is commonly used in commercial swine production to equalize litter sizes and/or adjust piglet birth weights within litters. However, there is limited published information on optimum cross-fostering procedures. This study evaluated the effects of within-litter birth weight variation after cross-fostering (using litters of 14 piglets) on piglet preweaning mortality (PWM) and weaning weight (WW). An RCBD was used (blocking factors were day of farrowing and sow parity, body condition score, and functional teat number) with an incomplete factorial arrangement of the following two treatments: 1) birth weight category (BWC): light (<1.0 kg), medium (1.0 to 1.5 kg), or heavy (1.5 to 2.0 kg); 2) litter composition: uniform, all piglets in the litter of the same BWC [uniform light (14 light piglets); uniform medium (14 medium piglets); uniform heavy (14 heavy piglets)]; mixed, piglets in the litter of two or more BWC [L+M (seven light and seven medium piglets); M+H (seven medium and seven heavy piglets); L+M+H (three light, six medium, and five heavy piglets)]. Piglets were weighed at 24 h after birth and randomly allotted to litter composition treatment from within BWC; all piglets were cross-fostered. There were 47 blocks of six litters (total 282 litters and 3,948 piglets). Weaning weights were collected at 18.7 ± 0.64 d of age; all PWM was recorded. Individual piglet WW and PWM data were analyzed using PROC MIXED and PROC GLIMMIX of SAS, respectively; models included fixed effects of BWC, litter composition, and the interaction, and random effects of sow within the block. There was litter composition by BWC interactions (P ≤ 0.05) for WW and PWM. Within each BWC, WW generally increased and PWM generally decreased as littermate weight decreased. For example, WW was greatest (P ≤ 0.05) for light piglets in uniform light litters, for medium piglets in L+M litters, and for heavy piglets in L+M+H litters. Preweaning mortality was lowest (P ≤ 0.05) for medium piglets in L+M litters, and for heavy piglets in L+M+H litters; however, litter composition had no effect (P > 0.05) on PWM of light piglets. In conclusion, increasing the average birth weight of littermates after cross-fostering generally decreased WW and increased PWM for piglets of all birth weight categories. This implies that the optimum approach to cross-fostering that maximizes piglet preweaning growth and survival is likely to vary depending on the birth weight distribution of the population.