Maternal Restricted- and Over-Feeding During Gestation Result in Distinct Lipid and Amino Acid Metabolite Profiles in the Longissimus Muscle of the Offspring

Abstract

Maternal over- and restricted-feeding during gestation have similar negative consequences for the offspring, including decreased muscularity, increased adiposity, and altered metabolism. Our objective was to determine the effects of poor maternal nutrition during gestation (over- and restricted-feeding) on the offspring muscle metabolite profile. Pregnant ewes (n = 47) were fed 60% (RES), 100% (CON), or 140% (OVER) of NRC requirements starting at day 30.2 ± 0.2 of gestation. Offspring sample collection occurred at days 90 and 135 of gestation, and within 24 h of birth. C2C12 myoblasts were cultured in serum collected from offspring at birth (n = 18; 6 offspring per treatment) for analysis of oxidative and glycolytic capacity. Unbiased metabolite analysis of longissimus muscle samples (n = 72; 8 fetuses per treatment per time point) was performed using mass spectrometry. Data were analyzed by ANOVA for main effects of treatment, time point, and their interaction. Cells cultured in serum from RES offspring exhibited increased proton leak 49% (p = 0.01) compared with CON, but no other variables of mitochondrial respiration or glycolytic function were altered. Mass spectrometry identified 612 metabolites. Principle component analysis identified day of gestation as the primary driver of metabolic change; however, maternal diet also altered the lipid and amino acid profiles in offspring. The abundance of 53 amino acid metabolites and 89 lipid metabolites was altered in RES compared with CON (p ≤ 0.05), including phospholipids, sphingolipids, and ceramides within the lipid metabolism pathway and metabolites involved in glutamate, histidine, and glutathione metabolism. Similarly, abundance of 63 amino acid metabolites and 70 lipid metabolites was altered in OVER compared with CON (p ≤ 0.05). These include metabolites involved in glutamate, histidine, lysine, and tryptophan metabolism and phosphatidylethanolamine, lysophospholipids, and fatty acids involved in lipid metabolism. Further, the amino acid and lipid profiles diverged between RES and OVER, with 69 amino acid and 118 lipid metabolites differing (p ≤ 0.05) between groups. Therefore, maternal diet affects metabolite abundance in offspring longissimus muscle, specifically metabolites involved in lipid and amino metabolism. These changes may impact post-natal skeletal muscle metabolism, possibly altering energy efficiency and long-term health.

Keywords: gestation; maternal diet; metabolism; muscle; offspring.

FIGURE 1

Principle component analysis (PCA) and hierarchical clustering obtained from employing metabolic concentration of all metabolites detected in longissimus muscle (LM). PCA of LM samples indicates that time is the primary driver of variance between samples. LM samples (n = 72; 8 per treatment per time point) were collected from offspring born to ewes fed control- (100%), restricted- (60%), or over-fed (140%) diets at days 90 or 135 of gestation, or within 24 h of birth were analyzed by UPLC-MS/MS. Offspring from control-, restricted-, or over-fed ewes are denoted as CON, RES, and OVER, respectively.

FIGURE 2

Poor maternal diet alters relative abundance of lipid (A) and amino acid (B) metabolites in the offspring longissimus muscle in a diet and time dependent manner. Offspring from ewes fed a control (CON; 100% NRC), over-fed (OVER; 140% NRC), or restricted-fed (RES; 60% NRC) diet were sampled at days 90 or 135 of gestation, or within 24 h of birth. Identified names are metabolite sub-pathways. Cells represent fold changes of abundance by color range, from red (row max) to white (row average) and blue (row minimum) between treatment groups.

FIGURE 3

Poor maternal diet results in distinct lipid and amino acid metabolite profiles in offspring longissimus. Offspring from ewes fed a control (CON; 100% NRC), over-fed (OVER; 140% NRC), or restricted-fed (RES; 60% NRC) diet were sampled at days 90 or 135 of gestation, or within 24 h of birth. Metabolite sub-pathways increased (red) or decreased (blue) in abundance at each time point in RES vs. CON (A) or OVER vs. CON (B). BCAA, branched chain amino acid; SAM, S-adenosyl methionine.