Maternal sleep deprivation disrupts glutamate metabolism in offspring rats

Abstract

Maternal sleep deprivation (MSD) has emerged as a significant public health concern, yet its effects on offspring metabolism remain poorly understood. This study investigated the metabolomic implications of MSD on offspring cognitive development, with a particular focus on alterations in glutamate metabolism. Pregnant rats were subjected to sleep deprivation during late gestation. Plasma and brain samples from their offspring were collected at different postnatal days (P1, P7, P14, and P56) and analyzed using untargeted metabolomics with liquid chromatography-mass spectrometry. Metabolomic analysis revealed significant differences in various amino acids, including L-glutamate, L-phenylalanine, L-tyrosine, and L-tryptophan, which are crucial for cognitive function. Subsequent differential analysis and partial least squares discriminant analysis (sPLS-DA) demonstrated a gradual reduction in these metabolic differences in the brain as the offspring underwent growth and development. KEGG pathway analysis revealed differential regulation of several pathways, including alanine, aspartate, and glutamate metabolism, glutathione metabolism, arginine biosynthesis, aminoacyl-tRNA biosynthesis, histidine metabolism, and taurine and hypotaurine metabolism, at different developmental stages. Mantel and Spearman analyses indicated that the observed changes in metabolites in MSD progeny may be related to various gut microbes, Ruminococcus_1, Ruminococcaceae_UCG-005, and Eubacterium_coprostanoligenes_group. Biochemical assays further demonstrated developmental changes in the L-glutamate metabolic pathway. Collectively, these findings suggest that MSD not only affects maternal well-being but also has enduring metabolic consequences for offspring, particularly impacting pathways linked to cognitive function. This highlights the importance of addressing maternal sleep health to mitigate potential long-term consequences for offspring.

Keywords: Cognitive development; Glutamate metabolism; Maternal sleep deprivation; Metabolomics; Offspring.

Figure 1

Animal model and sampling Pregnant rats were exposed to MSD for 6 h daily during gestation days 15–21, with prior acclimation to gentle handling procedures. Plasma, brain tissue, and intestinal content samples were collected from offspring on postnatal days 1, 7, 14, and 56 (P1, P7, P14, and P56, respectively).

Figure 2

Untargeted metabolomic analysis of plasma samples from offspring at different developmental stages A–C: Volcano plot of differential metabolites between MSD and control offspring at P7 (A), P14 (B), and P56 (C). D–F: sPLS-DA of metabolomes between MSD and control offspring at P7 (D), P14 (E), and P56 (F). G: Venn diagram of overlapping differential metabolites between MSD and control offspring at different time points in plasma. H: VIP scores of 27 common differential metabolites identified in the Venn diagram.

Figure 3

Untargeted metabolomic analysis of brain samples from offspring at different developmental stages A–D: Volcano plot of differential metabolites between MSD and control offspring at P1 (A), P7 (B), P14 (C), and P56 (D). E–H: sPLS-DA of metabolomes between MSD and control offspring at P1 (E), P7 (F), P14 (G), and P56 (H). I: Venn diagram of differential metabolites between MSD and control groups at different time points in the brain. J: VIP scores of 36 common differential metabolites identified in the Venn diagram.

Figure 4

sPLS-DA and pathway analysis of MSD and control offspring A–B: sPLS-DA plots of metabolomes in control (A) and MSD plasma (B) across all developmental stages. C: Graphical representation of pathway analysis of plasma samples (FDR<0.05). D, E: sPLS-DA plots of metabolomes in control (D) and MSD brain (E) across all developmental stages. F: Graphical representation of pathway analysis of brain samples (FDR<0.05).

Figure 5

Analysis of relationship between metabolites and gut microbiota A: Mantel analysis of correlation between common differential metabolites in plasma and brain with gut microbiota. B, C: Spearman analysis of correlation between identified gut microbiota from Mantel test and common differential metabolites in plasma (B) and brain samples (C). ^*: P<0.05; ^**: P<0.001; ^***: P<0.0001. D: Venn diagram showcasing common differential metabolites in both brain and plasma. E, F: Comparative analysis of concentrations of five common differential metabolites in plasma (E) and brain samples (F) between MSD and control groups. G–J: Dots, with colors corresponding to AUCs depicted in Supplementary Figures S3, S4, signify areas under the ROC curves for metabolites exhibiting biomarker potential for MSD-exposed offspring. Ctrl: Control group. MSD: Maternal sleep deprivation group.

Figure 6

Biochemical assessment of glutamate, glutathione, and cysteine levels A–C: Glutamate concentrations in plasma (A), cortex (B), and hippocampus (C). D–F: Glutathione concentrations in plasma (D), cortex (E), and hippocampus (F). G–I: Cysteine concentrations in plasma (G), cortex (H), and hippocampus (I). Glu: Glutamate; GSH: Glutathione; Cys: Cysteine. Ctrl: Control group. MSD: Maternal sleep deprivation group. ^*: P<0.05; ^**: P<0.001.

Figure 7

Overview of glutamate and glutathione metabolism GSSG: Glutathione disulfide; GABA: Gamma-aminobutyric acid; GPX4: Glutathione peroxidase 4; GR: Glutathione reductase; GCL: γ-glutamylcysteine ligase; GFAT: Glutamine fructose-6-phosphate amidotransferase; GAD: Glutamate decarboxylase; TCA: Tricarboxylic acid cycle.