Search Results (287)

Background: Early-life sensory experiences play a crucial role in the maturation of neural circuits underlying sensorimotor development. The present study investigated whether the effects of neonatal tactile stimulation (NTS) on sensorimotor development depend on the developmental timing of the intervention in WAG/Rij rat pups, a well-established genetic model of absence epilepsy. Methods: Seventy rat pups were randomly assigned to five groups: Wistar control, WAG/Rij control, and WAG/Rij rats receiving NTS during the first (PN Week 1-NTS), second (PN Week 2-NTS), or third (PN Week 3-NTS) postnatal week. Tactile stimulation was administered three times daily. Sensorimotor development was assessed on postnatal day 22 using a modified rung-bridge task that evaluated orientation, flexor/extensor activity, distal control, gait development, postural control, sensorimotor responses, tail use, total sensorimotor performance, and crossing time. Results: WAG/Rij control pups exhibited impaired sensorimotor performance compared with Wistar controls, demonstrating lower distal control, gait development, sensorimotor response, and total sensorimotor scores, together with prolonged crossing times. The effects of the NTS protocol were dependent on the developmental window in which it was administered. The NTS protocol applied during the second postnatal week was associated with the most pronounced improvements, including significantly higher distal control, sensorimotor response, tail performance, and total sensorimotor scores, as well as significantly shorter crossing times compared with untreated WAG/Rij controls. In contrast, orientation, flexor/extensor activity, and postural control were not significantly affected. Discussion and Conclusions: These findings indicate that sensorimotor abnormalities are detectable in WAG/Rij rats during the early postnatal period, before the typical adult expression of absence seizures. The effects of the NTS protocol were strongly dependent on developmental timing, with the second postnatal week representing a particularly sensitive developmental window for sensorimotor maturation. Overall, the NTS protocol was associated with improved early sensorimotor performance in WAG/Rij rats, particularly when administered during the second postnatal week. Because a sham-handled control group was not included, these findings should be interpreted as reflecting the NTS protocol as a whole rather than tactile stimulation alone. Full article

Chronic lung disease of prematurity (CLD) is a common complication of preterm birth with a complex pathology. Recent epidemiologic studies have identified a link between neonatal exposure to di(2-ethylhexyl) phthalate (DEHP), frequently used in medical equipment, and the development of CLD. We hypothesize that DEHP exposure in the early neonatal period contributes to lung injury in newborn rats. Newborn rat pups were raised in one of the following environments: room air (RA), RA + DEHP, hyperoxia (60% oxygen), and hyperoxia + DEHP. Ambient DEHP was inhaled at a dose of 25 mg/m³ for 6 h daily for 14 days. Lung tissue and blood samples were collected on the 14th day of life. Independent exposure to DEHP and hyperoxia resulted in thicker pulmonary septal walls, fewer alveoli, increased pulmonary polymorphonuclear leukocytes and myeloperoxidase (MPO) activity and decreased expression of CD31 on endothelial cells in lung tissue. Additionally, DEHP-exposed rats showed higher serum malondialdehyde (MDA) levels and reduced vascular endothelial growth factor (VEGF) mRNA and protein levels compared to controls. Our experiments demonstrate that inhaled DEHP, with or without hyperoxia, resulted in a similar pattern of morphological lung injury and inflammation characteristic of CLD, suggesting an association with CLD of prematurity. Full article

Background and Objectives: Green tea is known for its powerful antioxidant properties. However, the effects of green tea consumption during pregnancy on neonatal development and the mechanisms of these effects are not fully understood. The aim of this study was to investigate potential damage to atrial cardiomyocytes of newborn rat pups whose mothers received green tea during pregnancy and to elucidate the apoptotic mechanisms underlying this possible damage. Materials and Methods: Wistar albino rats (weighing 200–220 g, 10 weeks old) were used in this study. Following the confirmation of pregnancy, rats were randomly assigned to groups, and the experimental group was administered green tea by oral gavage at a dose of 50 mg/kg per day for 21 days. Atrial cardiomyocytes and mitral valve cells from newborn pups (postnatal day 1) were obtained and evaluated immunohistochemically for cytochrome c, caspase-9, and caspase-3 expression. Results: TUNEL analysis revealed a significant increase in DNA fragmentation in the green tea group, with the median number of apoptotic cells per region of interest (ROI) rising from 5.5 to 24.5 in atrial cardiomyocytes (p < 0.001), and from 2.0 to 10.0 in mitral valve cells (p < 0.05). Immunohistochemically, the control group showed faint-to-weak basal immunoreactivity of cytochrome c and caspase-3, and weak-to-moderate expression of caspase-9. In the green tea group, caspase-3 immunoreactivity was moderate, while cytochrome c and caspase-9 immunoreactivity were significantly higher. Quantitative HSCORE analysis confirmed significant elevations in atrial cardiomyocytes for cytochrome c (from 65.0 to 210.0; p < 0.001), caspase-9 (from 85.0 to 140.0; p < 0.001), and caspase-3 (from 60.0 to 120.5; p < 0.001). Similar statistically significant increases were observed across all corresponding markers in the mitral valve cells (p < 0.05). Overall, the induction of apoptosis was notably more pronounced in atrial cardiomyocytes than in mitral valve cells. Conclusions: Our findings suggest that the mechanism of potential damage in atrial cardiomyocytes of newborn rat pups is associated with mitochondria-mediated apoptosis, potentially triggered by activation of the cytochrome c, caspase-9 and caspase-3 axis. These results highlight the importance of exercising caution regarding the consumption of green tea supplements during pregnancy. Further studies are needed to correlate these preliminary neonatal observations with clinical outcomes. Full article

Background/Objectives: Neuritogenesis and synaptogenesis support learning and cognitive function, and hippocampal neurons play central roles in these processes. Cleistocalyx nervosum var. paniala (CNP), a Southeast Asian berry, has reported neuroprotective activities, but its direct effects on hippocampal neurons remain unclear. We investigated whether CNP extract modulates hippocampal neuronal transcriptomes, neuritogenesis, and synaptogenesis. Methods: Primary hippocampal neurons isolated from male and female Wistar rat pups were treated with CNP extract in vitro. Cytotoxicity was assessed to define non-cytotoxic concentrations. Transcriptomic responses were profiled by RNA sequencing and validated by RT-qPCR. Neuritogenesis was quantified by neurite morphology and Sholl analysis. Synaptogenesis was evaluated by synaptic immunocytochemistry. Molecular docking of cyanidin-3-glucoside (C3G) and resveratrol was used to generate mechanistic hypotheses. Results: At 0.1–10 µg/mL, CNP was non-cytotoxic, whereas a 100 µg/mL dose reduced viability; therefore, 10 µg/mL was used in subsequent experiments. Exploratory RNA-seq profiling identified thousands of differentially expressed genes enriched in synapse- and neurite-related pathways, including synaptogenesis signaling, axon guidance, and neuritogenesis. RT-qPCR showed upregulation of Igf1 in males and Glul in females, with sex-dependent modulation of Bdnf and Cask. CNP increased neurite length, branching, and Sholl complexity in both sexes, with a more pronounced effect in males. A male-biased effect was also observed in synapse-related marker colocalization, with increased Syn1–Psd95 colocalization detected in males. Docking suggested plausible interactions of C3G and resveratrol with regulators such as MYC, TP53, and CREB1. Conclusions: CNP extract alters transcriptional networks and enhances neurite outgrowth in primary hippocampal neurons in a sex-dependent manner, with male-biased effects on Syn1–Psd95 colocalization. These findings support further dose–response, mechanistic, and sex-stratified in vivo studies to evaluate its neurobiological potential. Full article

Background/Objectives: The vibrotactile system, which is essential for guiding behavior in nocturnal rodents such as mice and rats, provides critical sensory input. To investigate the role of vibrotactile sensory inflow in neonatal locomotion, we used previsual rat pups that underwent bilateral vibrissectomy. Subsequently, their motor behavior was evaluated in an open field test. Methods: A total of 42 previsual pups from four litters were assigned to either bilateral vibrissectomy or sham surgery groups on postnatal days (PND) 9–12, with group allocation balanced across litters. Results: Open-field testing on PND 13 revealed that while vibrissectomy (VE) did not affect gross locomotor activity—such as distance traveled, speed, acceleration, or freezing episodes (all >0.05)—it significantly altered spatial behavior. To quantify spatial patterns of curvy tracks, we analyzed trajectorial compaction within the central zone, lacking the tactile guidance of the walls: trajectories were smoothed using virtual coatings scaled to the vibrissal length (16 mm). For each track, an individual linearized reference path was generated and subjected to identical smoothing. The compaction ratio—calculated as the coated area of the smoothed linearized reference divided by the coated area of the experimental track—was significantly greater in VE pups than in sham controls (p = 0.03). This effect was not attributable to differences in the path length traveled within the central zone. The increased compaction persisted when the smoothing scale was increased 2–3 fold (32–64 mm radii, approximating the pups’ mean body size), but not at smaller scales (2–4 mm). Conclusions: These results demonstrate that tactile input specifically modulates the spatial, rather than locomotor, components of nonvisual navigation. Consequently, the track compaction may serve as a sensitive marker for assessing vibrotactile function in developing laboratory rodents. Full article

Background: Environmental agents can disrupt thyroid function at several levels, including the synthesis, action, and excretion of thyroid hormones, and an inadequate concentration of thyroid hormones affects almost all organs and systems. Objective: The present study aimed to evaluate the histology and presence of the cytokines TNFα, IL-6, and IL-10 in the thyroid gland by immunohistochemical labeling, as well as the body weight and craniocaudal length of pups of Wistar rats exposed to ambient air in the vicinity of the Capuava Petrochemical Complex (CPC), located in the Santo André and Mauá cities, at São Paulo State, Brazil. Methods: This study used Wistar rats between 14 and 16 weeks of age, distributed in couples, that were exposed to pollution from the CPC located in the regions of Santo André and Mauá, Sao Paulo State, Brazil. One couple was positioned 600 m (SS1), and another was 1000 m (SS2) from the CPC, while the control group was kept at the animal research facility of the Physiology Laboratory of the FMABC University Center, Santo André. After mating, the resulting offspring were monitored for four weeks, with their body weight and craniocaudal length measured weekly. Subsequently, the offspring’s thyroid glands were histologically analyzed using H&E staining and immunohistochemistry to detect the presence of inflammatory cytokines (TNF-α, IL-6, and IL-10). Results: In the SS1 group, the thyroid glands showed follicular heterogeneity with macrofollicles and numerous microfollicles without colloid, lined by flattened epithelial cells. In these thyroid follicles, there was intense TNFα (p = 0.002) staining, slight IL-6 staining (p = 0.042), and significantly stronger staining for IL-10 (p = 0.013) compared to that in the control group. This group also had a significantly lower body weight than the control animals on the 6th, 13th, and 20th days of life. In the SS2 group, the thyroids presented an architecture dominated by microfollicles without colloid as well as inflammatory cells in the colloid of some follicles. Immunohistochemistry revealed intense pan-follicular TNFα (p = 0.002) staining, with additional cytoplasmic staining of IL-6 (p = 0.040) and IL-10 (p = 0.006). The SS2 group also showed a more pronounced deficit compared to the SS1 group in terms of birth weight. The cranial–caudal length was shorter on the 13th and 20th days of life in the SS1 and SS2 groups compared to the control group. Conclusions: The results indicate that proximity of rats to the CPC was a determining factor in the development of histological abnormalities and increases in inflammatory cytokine markers in the thyroid glands of the offspring. In addition, the offspring born near the CPC had lower birth weights and shorter craniocaudal lengths compared to the animals in the control group. Full article

Hypoprolactinemia (hypoPRL) disrupts lactation and compromises milk production. Although maternal milk is a critical source of nutrients and bioactive compounds for newborns, the consequences of deficient lactation based on reduced milk quantity on the offspring’s immune development remain incompletely understood. Therefore, this study aimed to elucidate how deficient lactation due to hypoPRL interferes with offspring immunity and development. Female Sprague Dawley (SD) and spontaneous hypoPRL Oncins France Colony A (OFA) rats were euthanized on day 2 of lactation to assess the impact of hypoPRL on serum, milk, and tissue samples. We demonstrated that early deficient lactation in the OFA model impaired maternal performance, leading to increased pup mortality during early lactation. OFA pups exhibited reduced body weight and weight gain, decreased cerebral weight and index, and an increased cephalization index. Thymic development was markedly altered, as evidenced by reduced thymus weight, area, and cortical extension. These structural changes were accompanied by increased thymic Rag 1 expression, suggesting altered lymphocyte development. In parallel, passive immune transfer was compromised, with reduced levels of OVA-specific immunoglobulin isotypes detected in pup serum, reflecting changes in milk-derived immune support. In summary, maternal hypoPRL during early lactation adversely affects offspring growth and thymic maturation with lasting consequences for neonatal immune maturation. This study provides a novel experimental framework to investigate the consequences of lactation deficiency with potential implications for understanding lactation insufficiency and its impact on neonatal immune exposure. Full article

Apelin is a postnatal peptide implicated in gastrointestinal maturation, yet its combined effects on mucosa, enteric plexuses, and gut-derived appetite signals are not well defined. We investigated the impact of chronic intragastric apelin-13 on the stomach and small intestine of unweaned rats. Twelve Wistar pups of both sexes received apelin-13 (100 nmol/kg body weight, twice daily) or saline from postnatal day 10 for 14 days. After euthanasia, gastric and small-intestinal samples were processed for histomorphometry, neurofilament immunohistochemistry of myenteric and submucosal plexuses, and quantitative staining for ghrelin and leptin. Apelin-13 increased gastric mucosal thickness and pit and gland height, enlarged zymogen cells, and reduced muscularis propria thickness, while leaving submucosa and parietal cell area unchanged. In the small intestine, apelin produced a clear proximal-distal gradient, with enhanced villus-mucosa indices proximally and reduced indices in mid-to-distal jejunum, alongside broader crypt remodeling. Enterocyte and goblet cell dimensions changed in parallel with these regional shifts. Myenteric and submucosal ganglia were also remodeled in a segment-dependent manner. Ghrelin immunoreactivity increased in most regions, whereas leptin showed opposite proximal and distal responses. Overall, early-life luminal apelin-13 reshapes gastric and intestinal architecture and local hormone expression. Full article

Necrotizing enterocolitis (NEC) is a life-threatening inflammatory disease of preterm infants, increasingly viewed as a cytokine-driven disorder of the immature intestine. We aimed to characterize local peritoneal concentrations of interleukin (IL)-1β, IL-6 and tumor necrosis factor-α (TNF-α) in a standardized neonatal rat NEC model and relate them to histopathological injury. Seventy-four SPRD/Mol/Lodz rat pups were allocated to a control group (CTRL; n = 12) or subjected to a hypoxia-hypothermia-formula-feeding NEC protocol (NEC; n = 62). After 72 h, small-intestinal samples were scored using a four-tier NEC scale (0–3), and peritoneal fluid cytokine levels were measured by ELISA. All CTRL animals exhibited normal histology (grade 0), whereas NEC pups showed a wide spectrum of lesions, with 66.6% classified as grade 2–3 and a significantly higher mean NEC score in NEC than CTRL (p < 0.001). Peritoneal IL-1β and TNF-α concentrations were markedly elevated in NEC versus CTRL animals (both p < 0.001), while IL-6 levels showed no statistically significant between-group difference. These findings indicate that experimental NEC in this model is accompanied by a pronounced local pro-inflammatory response dominated by IL-1β and TNF-α, whereas IL-6 may follow distinct temporal or compartment-specific kinetics. Peritoneal cytokine profiling may help refine mechanistic understanding and guide future biomarker and immunomodulatory strategies in NEC. Full article

β-casein (β-CN) is the predominant casein fraction in breast milk, while current infant milk formula (IMF) contains substantially lower β-CN levels than breast milk. The impact of β-CN fortification on neonatal digestive characteristic and bioactive peptide release remains an understudied area in vivo. This study investigated the effect of β-CN fortification in milk protein on digestion properties and release of bioactive peptides using a suckling rat pup model. Rat pups were, respectively, gavaged with two milk protein solutions: one with ordinary β-CN content (OBCN) and the other with fortified β-CN content (FBCN). The gastric emptying rate, proteolytic efficiency, and peptidomic profiles of intestinal digesta were evaluated. Results indicated that the FBCN group exhibited accelerated gastric emptying into the intestinal phase and enhanced proteolytic efficiency compared to OBCN group. Furthermore, the FBCN group generated greater peptide diversity in the small intestine, with significantly elevated abundance of bioactive peptide candidates exhibiting broader functional spectra. These findings provide additional evidence for the health effects of β-CN fortification in IMF. Full article

Fetal alcohol spectrum disorder (FASD/FAS) is a chronic inflammatory process of the fetal brain induced by alcohol and mediated by pro-inflammatory (PILM) and pro-resolving (PRLM) lipid mediators of inflammation. DHA (docosahexaenoic acid) is an essential precursor of PRLM. A study examining the response of lipid mediators of inflammation to alcohol insult and DHA supplementation can provide vital information on the pathogenesis of FASD/FAS and the potential ameliorative role of DHA. Four groups of timed pregnant rats were studied: control, low-dose (1.6 g/kg/day) and high-dose (2.4 g/kg/day) alcohol, and high-dose alcohol (2.4 g/kg/day) + DHA (1250 mg/kg/day). The pups were delivered on day 20, and their whole brain was examined for lipid mediators by liquid chromatography mass spectroscopy. The following biomarkers of brain lipid mediators were studied, namely, PILM (LTB4, PGE2, PGF2α, TXB2) and PRLM (LXA5, 4-HDoHE, 17-HDoHE, and MaR1n-3, DPA). The brain PILM and PRLM concentrations decreased significantly (p < 0.001) with high-dose alcohol. However, high-dose alcohol + DHA resulted in a significant (p < 0.001) increase in PRLM levels, viz., LXA5, MaR1n-3 DPA, 17-HDoHE, and a threefold increase in 4-HDoHE. We conclude that DHA supplementation in alcohol-exposed pregnant rats significantly increased levels of brain pro-resolving lipid mediators in the offspring, suggesting a potential role in modulating the inflammatory response. Full article

Background: Metabolic memory refers to the long-term adverse effects of short-term disturbances in glucose metabolism. Recent evidence indicates that hyperglycemia-induced metabolic memory contributes to sustained cellular damage even after glycemic control, driven by increased production of reactive oxygen species (ROS), activation of inflammatory pathways, and accumulation of advanced glycation end products (AGEs). Although well characterized in endothelial and smooth muscle cells, this phenomenon may also occur in other cell types, including glial cells. Objective: This study aimed to evaluate the persistence of high-glucose (HG)-induced alterations after returning to normal glucose (NG) conditions in primary mixed glial cell (MGC) cultures. Methods: Primary MGCs were obtained from neonatal Wistar rat pups and cultured under three conditions for 21 days: NG (5.5 mM glucose), HG (25 mM glucose), and HG-NG (14 days in HG followed by 7 days in NG). Cell proliferation, apoptosis, ROS production, lipid peroxidation, mitochondrial activity, TNF-α, IL-6, and AGE formation were assessed. Results: MGCs cultured under HG and HG-NG conditions exhibited reduced proliferation without increased apoptosis. Both HG and HG-NG conditions promoted ROS overproduction accompanied by reduced mitochondrial activity, whereas only HG increased lipid peroxidation. Notably, TNF-α and AGE levels were elevated in both HG and HG-NG conditions, while IL-6 production decreased exclusively in HG-NG. Conclusions: These findings demonstrate the persistence of deleterious effects induced by HG in MGCs, even after restoration to NG conditions. Full article

Background/Objectives: Overnutrition increases comorbidities such as gestational diabetes during pregnancy that can have detrimental consequences for both parent and progeny. We previously reported that high-fat (HF) diet and late-gestation diabetes (DM) incite mitochondrial dysfunction, oxidative stress, and cardiometabolic disease in first generation (F1) rat offspring, partially through epigenomic and transcriptomic programming. Primordial germ cells, which become the second generation (F2), are also exposed, which could incite generational risk. This study aimed to determine whether the F2 transcriptome already has genomic variation at the preimplantation embryo stage, and whether variations normalize, persist or compound in the third generation (F3). Methods: F0 female rats were fed a control or HF diet, then DM was induced in HF-fed dams on gestational day (GD)14, exposing F1 offspring and F2 primordial germ cells to hyperlipidemia, hyperglycemia and fetal hyperinsulinemia during the last third of pregnancy. F1 pups were reared by healthy dams and bred to produce F2 embryos (F2e) and F2 pups. F2 offspring were bred to produce F3 embryos (F3e). Embryos were assessed by a novel grading method, live cell imaging, and single-cell RNA sequencing. Results: Embryo grades were not different, but HF+DM F2e had more cells while F3e had fewer cells and overall fewer embryos. HF+DM F2e had similar mitochondria quantity but a downregulation of genes involved in lipid metabolism and more oxidative stress, consistent with mitochondrial dysfunction. They also had an upregulation of chromatin-remodeling genes. The predicted developmental effect is accelerated embryo aging and epigenetic drift. In contrast, HF+DM F3e had an adaptive stress response leading to increased mitochondria quantity and an upregulation of genes involved in mitochondrial respiration, metabolism, and genomic repair that led to a predicted developmental effect of delayed embryo maturation. Conclusions: Although pathways vary, both generations have metabolically linked differentially expressed genes that influence cell fate and developmental pathways. In conclusion, HF+DM pregnancy can program the early embryonic transcriptome for three generations, despite an intergenerational healthy diet. Full article

Maternal diet influences offspring development, immune function, and intestinal health. This study investigates the effects of maternal supplementation with a key component of the Mediterranean Diet, extra virgin olive oil (EVOO), on the immune health of offspring at the end of lactation. Lewis rat dams received either refined olive oil (ROO), EVOO, or water (REF) during gestation and lactation. Plasma immunoglobulin G2c (IgG2c) concentration was elevated in pups born to EVOO-supplemented mothers, indicating enhanced immune development. Histological analysis of the small intestine revealed more goblet cells in the EVOO group, indicating a potential positive effect on the intestinal barrier function. In vitro assays showed that EVOO metabolites did not display cytotoxicity and had improved barrier integrity under a stress stimulus. These findings suggest that maternal EVOO supplementation may have beneficial effects on immune and intestinal development and health in offspring. Full article

Background/Objectives: The hippocampus is an essential part of the central nervous system (CNS); it plays a significant role in social–cognitive memory processing. Prenatal exposure to valproic acid (VPA) can lead to impaired hippocampal functions. In this study, we evaluated the effect of plumbagin (PLB) as a natural product on spatial learning and memory, neuro-morphological changes, and inflammation levels in a VPA-induced autism model during adolescence. Methods: Pregnant Wistar rats received a single intraperitoneal (i.p.) injection of VPA (600 mg/kg) or saline on gestational day 12.5. The male offspring were then categorized and assigned to five groups: Saline+DMSO-, VPA+DMSO-, and VPA+PLB-treated groups at doses of 0.25, 0.5, or 1 mg/kg. Spatial learning and memory were evaluated using the Morris water maze. Histopathological evaluations of the hippocampus were performed using Nissl and hematoxylin–eosin staining, as well as immunofluorescence. The pro-inflammatory cytokine levels were also quantified by quantitative real-time PCR. Results: The findings revealed that a VPA injection on gestational day 12.5 is associated with cognitive impairments in male pups, including a longer escape latency and traveled distance, as well as decreased time spent in the target quadrant. Treatment with PLB significantly enhanced the cognitive function, reduced dark cells, and ameliorated neuronal–morphological alterations in the hippocampus of VPA-exposed rats. Moreover, PLB was found to reduce astrocyte activation and the expression levels of pro-inflammatory cytokines. Conclusions: These findings suggest that PLB partly mitigates VPA-induced cognitive deficits by ameliorating hippocampal inflammation levels. Full article