Maternal Nutrient Restriction Research Articles

PSVIII-3 Maternal nutrient restriction during mid-to-late gestation alters adipogenic gene expression in subcutaneous and perirenal adipose tissue of fetal beef cattle offspring

Abstract Proper maternal nutrition during gestation is an important determinant of fetal and long-term developmental programming. Transcriptional networks involved in economically important processes such as adipogenesis and overall growth of the offspring are particularly susceptible to developmental perturbations and programming. Low quality forages or insufficient maternal nutrient supplementation during gestation are scenarios that can negatively affect fetal growth and development by altering the regulation of these networks. Therefore, the purpose of this study was to determine the effect of maternal nutrient restriction during mid-to-late gestation on mRNA expression of genes involved in growth and adipogenesis in fetal back fat (BF), and perirenal adipose tissue (PR). A total of six multiparous, commercial cows were equally divided into one of two dietary treatments: Control (CON; fed 100% of NRC recommendations) vs. Restricted Feed (RES; fed 60% of NRC recommendations) from 140 to 240 days of gestation. The animals were euthanized on day 240 of gestation for fetal tissue collection and qPCR analysis using TaqMan® gene expression assays. Relative mRNA quantification was determined using the Delta-Delta Ct method. Differences between Least Square Means were compared using the PDIFF option of the MIXED procedure of SAS (9.4). PPARg expression was increased (P = 0.009) in BF tissue in the RES group compared to the CON group. IGF-1 expression tended to increase (P = 0.076), and IGF-1R expression tended to decrease (P = 0.055) in fetal BF. In PR tissue, CEBPA and IGF-1 expression decreased (P ≤ 0.043) in RES compared to CON. These findings suggest that as a result of a compromised fetal environment, the PR and BF depots are differentially regulated in a manner that may persist into postnatal life and warrants further investigation.

340 Effects of nutrient restriction during mid- to late-gestation on maternal and fetal pancreatic exocrine function in sheep

Abstract To examine the effects of nutrient restriction on fetal and maternal ovine exocrine pancreatic function during mid- to late-gestation, 41 singleton ewes (48.3±0.6 kg BW) were randomly assigned to dietary treatments: 100% (control; CON; n = 20) or 60% of nutrient requirements (restricted; RES; n = 21) from day 50–90 (mid-gestation). At day 90, 14 ewes (CON, n = 7; RES, n = 7) were slaughtered. The remaining ewes were subjected to treatments of nutrient restriction or remained under a control diet from day 90–130 (late-gestation): CON-CON (n = 6), CON-RES (n = 7), RES-CON (n = 7), and RES-RES (n = 7) and were slaughtered at day 130. The pancreas was weighed, subsampled, and assayed for digestive enzyme activity. Enzyme activity was expressed as U/g, U/g protein, or U/pancreas (total content). Total protein content in the pancreas was expressed as g/pancreas. Differences between means were determined using contrasts in the MIXED procedure of SAS. Fetal and maternal pancreatic mass increased (P < 0.04) with day of gestation. Nutrient restriction during mid- (P = 0.01) and late-gestation (P = 0.01) decreased maternal pancreatic mass on d 130. Total fetal pancreatic α-amylase, trypsin, and protein content increased (P < 0.05) with day of gestation. Maternal nutrient restriction during late-gestation tended to decrease (P < 0.08) fetal pancreatic trypsin activity. Nutrient restriction during late-gestation tended to increase (P = 0.07) α-amylase:trypsin in fetal pancreas but tended to have the opposite response in maternal pancreas (P = 0.10). Total maternal pancreatic content of trypsin and protein tended to increase (P < 0.07) as gestation progressed. Nutrient restriction of gestating ewes decreased total content of a-amylase (P = 0.04) and tended to decrease total content of trypsin (P = 0.06) and protein (P = 0.06) in the maternal pancreas on d 90. Nutrient restriction during mid-gestation on d 90 and during late-gestation on d 130 decreased (P = 0.04) maternal α-amylase activity. Maternal nutrient restriction impairs pancreatic exocrine function by reducing maternal and fetal digestive enzyme activity.

345 The effects of maternal nutrient restriction followed by re-alimentation on offspring growth and metabolism in sheep

Abstract The duration and timing of inadequate maternal nutrition can have detrimental effects on metabolism and organogenesis in the offspring. Re-alimentation, a common management practice that involves feeding full nutrient requirements following a period of nutrient restriction, may reduce the negative impacts of maternal nutrient restriction. To determine the effects of maternal nutrient restriction and re-alimentation on offspring growth,48 primiparous ewes, confirmed pregnant with singletons, were fed a control diet consisting of100% NRC requirements (CON) starting on day25 of gestation. On day50 of gestation, ewes (n = 7) were euthanized and fetal liver, muscle, and blood samples were collected. The remaining animals were fed either CON or60% NRC requirements (RES). On day90 of gestation, a portion of ewes were euthanized (n = 7 per treatment) and fetal samples and weights were collected. Remaining ewes were maintained on the current diet (CON-CON, n = 6; RES-RES, n = 7) or switched to the alternative diet (CON-RES, RES-CON; n = 7/treatment). On day130 of gestation, all remaining ewes were euthanized. All fetal BW, liver, longissimus muscle, semitendinosus, and triceps brachii weights were determined for each day of gestation. Fetal BW’s were not different between treatment groups (P = 0.29; P = 0.83). Fetal liver weights decreased12.89% in RES-RES compared with CON-CON at day130 (P = 0.049), but were not different at day90 (P = 0.69). There was a tendency for decreased semitendinosus weight in RES group compared with CON at day90 (P = 0.055). Liver lipid droplet accumulation was analyzed for day90 and130 using histochemistry and an effect of maternal nutrition was not observed (P = 0.562). In summary, maternal nutrient restriction reduces offspring muscle and liver growth. To gain insight into the effects of maternal nutrient restriction and re-alimentation on liver development and metabolism, analysis of liver morphology, gene expression, and global metabolomics are needed. Supported by USDA-AFRI grant2016-67016-24884

PSIV-1 Effects of nutrient restriction and realimentation of gestating ewes on fetal carbohydrase activities in the small intestine

Abstract The small intestine plays an important role in post-ruminal carbohydrate digestion and there is limited information on its function in response to nutritional adaptation. To examine the effects of nutrient restriction and realimentation on fetal small intestinal carbohydrase activities during mid- to late-gestation, 41 singleton ewes (48.3±0.6 kg BW) were randomly assigned to dietary treatments: 100% (control; CON; n = 20) or 60% of nutrient requirements (restricted; RES; n = 21) from day 50–90 (mid-gestation) of gestation. At day 90, 14 ewes (CON, n = 7; RES, n = 7) were slaughtered. The remaining ewes were subjected to treatments of nutrient restriction or remained under a control diet from day 90–130 (late-gestation): CON-CON (n = 6), CON-RES (n = 7), RES-CON (n = 7), and RES-RES (n = 7) and were slaughtered at day 130. The fetal small intestine was weighed, subsampled, and assayed for carbohydrase (maltase, isomaltase, glucoamylase, lactase, and sucrase) activity. Enzyme activity was expressed as U/g and U/g protein. Protein concentration was expressed as mg/g intestine. Differences between means were determined using contrasts in the MIXED procedure of SAS. Small intestinal mass increased (P < 0.001) with day of gestation. Sucrase activity was undetected in the small intestine. Day of gestation did not affect carbohydrase development in the fetal small intestine. Nutrient restriction did not affect glucoamylase, maltase, or isomaltase activities. There was a significant mid-gestation treatment × late-gestation treatment interaction for lactase activity (P < 0.05). Realimentation during late-gestation after nutrient restriction during mid-gestation increased lactase activity in the fetal small intestine. These data demonstrate that fetal mucosal carbohydrases are imprinted early in gestation and brush border α-glycohydrolases involved in starch digestion (maltase, isomaltase, glucoamylase) do not respond to maternal nutrient restriction in sheep. Nutrient restriction of ewes during mid-gestation followed by realimentation during late-gestation may be a programming strategy to increase fetal lactase activity.

PSXIV-31 Effects of bovine maternal nutrient restriction on offspring microRNA and messengerRNA expression and muscle fiber type

Abstract Cows nutritionally restricted during mid-gestation produce calves with differential fat and muscle deposition when compared to calves from non-restricted cows. However, little is currently known about effects these changes have on post-natal skeletal growth. We investigated whether calves from cows restricted in the second trimester had different expression of microRNA (miR), messengerRNA, and muscle fiber type when compared to calves from non-restricted dams. Cows were bred by the same sire, stratified by body weight (P = 0.80) and allocated to one of two treatments: maintenance (n = 16) or restricted (n = 18). Restricted cows received lower forage biomass (1662 kg/ha, dry matter (DM)) compared to maintenance (2309 kg/ha, DM). Restricted cows had mean BCS 1.55 lower (P = 0.001) than maintenance cows and weight difference of 188 kg (P = 0.024). After the second trimester, all cows and calves were treated similarly. Skeletal muscle biopsies were collected from calves at weaning, beginning of the feedlot, and harvest. Offspring from restricted cows expressed more (P < 0.05) miR-133a, -133b, -181d, -214, -424 and -486 in the biceps femoris (BF) at the beginning of the feedlot when compared to maintenance offspring. At harvest, offspring from restricted cows expressed more (P < 0.05) miR-133a and less (P < 0.01) miR-486 in the longissimus lumborum (LD) than offspring from maintenance cows. There was no change (P ≥ 0.27) in expression of Pax3, Pax7, Cdc25A, MamL1, Ezh2, and IGF-1R between offspring from the two treatment groups in the BF at the beginning of the feedlot or the LD at harvest. Expression was not different (P ≥ 0.14) in myosin heavy chain isoforms MHC-1, -2a, and -2x between offspring from maintenance and restricted in the BF at the beginning of the feedlot or the LD at harvest. These data provide novel insight into the molecular mechanism through which nutrient restriction in the second trimester alters fat and muscle deposition in the offspring.

Maternal nutrient restriction in early pregnancy increases the risk of late embryo loss despite no effects on peri-implantation interferon-stimulated genes in suckler beef cattle

Reducing feeding costs in suckler beef herds to improve economic returns could have detrimental impacts on fertility. This study sought to determine whether maternal nutrient restriction during early pregnancy affects interferon-stimulated gene (ISG) expression in peripheral blood mononuclear cells during the peri-implantation period in two beef cattle breeds. Relationships were also examined between subnutrition and pregnancy failure defined according to ISG fold changes on Days 18 and 21 and to plasma pregnancy specific protein B (PSPB) concentrations on Day 28 post-artificial insemination (AI). Pirenaica or Parda de Montaña dams were assigned to a control (n = 23) or subnutrition (n = 30) group, receiving 100% or 65% of their estimated nutritional requirements from Day 1 to 82 post-AI, respectively. Treatment did not affect ISG expression or fertility. According to ISG fold changes (chi-square P = .023) or PSPB levels (chi-square P = .04) recorded in the subnutrition group, late embryo loss was more likely than in controls. Positive correlation was detected between Day 28 PSPB concentrations and both Day 18 MX1, MX2 and ISG15 expression, and Day 21 OAS1 expression. OAS1 and MX1 fold changes were found to be the best variables to discriminate pregnancy status. Our findings indicate that maternal nutrient restriction during the first third of pregnancy does not impair embryo signalling yet may increase the risk of pregnancy failure.

Insights image for "Evidence of increased hypoxia signalling in fetal liver from maternal nutrient restriction in mice".

Insights image for "Evidence of increased hypoxia signalling in fetal liver from maternal nutrient restriction in mice".

Change in Brain Plasmalogen Composition by Exposure to Prenatal Undernutrition Leads to Behavioral Impairment of Rats.

Epidemiological studies suggest that poor nutrition during pregnancy influences offspring predisposition to experience developmental and psychiatric disorders. Animal studies have shown that maternal undernutrition leads to behavioral impairment, which is linked to alterations in monoaminergic systems and inflammation in the brain. In this study, we focused on the ethanolamine plasmalogen of the brain as a possible contributor to behavioral disturbances observed in offspring exposed to maternal undernutrition. Maternal food or protein restriction between gestational day (GD) 5.5 and GD 10.5 resulted in hyperactivity of rat male adult offspring. Genes related to the phospholipid biosynthesis were found to be activated in the PFC, but not in the NAcc or striatum, in the offspring exposed to prenatal undernutrition. Corresponding to these gene activations, increased ethanolamine plasmalogen (18:0p-22:6) was observed in the PFC using mass spectrometry imaging. A high number of crossings and the long time spent in the center area were observed in the offspring exposed to prenatal undernutrition and were mimicked in adult rats via the intravenous injection of ethanolamine plasmalogen (18:0p-22:6) incorporated into the liposome. Additionally, plasmalogen (18:0p-22:6) increased only in the PFC, and not in the NAcc or striatum. These results suggest that brain plasmalogen is one of the key molecules to control behavior, and its injection using liposome is a potential therapeutic approach for cognitive impairment.SIGNIFICANCE STATEMENT Maternal undernutrition correlates to developmental and psychiatric disorders. Here, we found that maternal undernutrition in early pregnancy led to hyperactivity in rat male offspring and induced gene activation of phospholipid-synthesizing enzyme and elevation of ethanolamine plasmalogen (18:0p-22:6) level in the PFC. Intravenous injection of ethanolamine plasmalogen (18:0p-22:6) incorporated into the liposome maintained crossing activity and the activity was circumscribed to the center area for a long time period, as in prenatally undernourished offspring with aberrant behavior. Furthermore, the amount of ethanolamine plasmalogen (18:0p-22:6) increased in the PFC of the rat after injection. Our result suggests that brain plasmalogen is one of the key molecules to control behavior and that its injection using liposome is a potential therapeutic approach for cognitive impairment.

Evidence of increased hypoxia signaling in fetal liver from maternal nutrient restriction in mice.

Intrauterine growth restriction (IUGR) is a pregnancy condition where fetal growth is reduced, and offspring from IUGR pregnancies are at increased risk for type II diabetes as adults. The liver is susceptible to fetal undernutrition experienced by IUGR infants and animal models of growth restriction. This study aimed to examine hepatic expression changes in a maternal nutrient restriction (MNR) mouse model of IUGR to understand fetal adaptations that influence adult metabolism. Liver samples of male offspring from MNR (70% of ad libitum starting at E6.5) or control pregnancies were obtained at E18.5 and differential expression was assessed by RNAseq and western blots. Forty-nine differentially expressed (FDR < 0.1) transcripts were enriched in hypoxia-inducible pathways including Fkbp5 (1.6-fold change), Ccng2 (1.5-fold change), Pfkfb3 (1.5-fold change), Kdm3a (1.2-fold change), Btg2 (1.6-fold change), Vhl (1.3-fold change), and Hif-3a (1.3-fold change) (FDR < 0.1). Fkbp5, Pfkfb3, Kdm3a, and Hif-3a were confirmed by qPCR, but only HIF-2a (2.2-fold change, p = 0.002) and HIF-3a (1.3 p = 0.03) protein were significantly increased. Although a moderate impact, these data support evidence of fetal adaptation to reduced nutrients by increased hypoxia signaling in the liver.

Relationship between maternal global nutrient restriction during pregnancy and offspring kidney structure and function: a systematic review of animal studies

Maternal undernutrition during pregnancy is prevalent across the globe, and the origins of many chronic diseases can be traced back to in utero conditions. This systematic review considers the current evidence in animal models regarding the relationship between maternal global nutrient restriction during pregnancy and offspring kidney structure and function. CINAHL, Cochrane, EMBASE, MEDLINE, and Scopus were searched to November 2017. Preferred Reporting Items for Systematic Reviews and Meta-analysis guidelines were followed, and articles were screened by two independent reviewers. Twenty-eight studies met the inclusion criteria: 16 studies were on rats, 9 on sheep, 2 on baboons, and 1 on goats. The majority of the rat studies had maternal global nutrient restriction during pregnancy at 50% of ad libitum while restriction for sheep and baboon studies ranged from 50% to 75%. Because of the heterogeneity of outcome measures and the large variation in the age of offspring at followup, no meta-analysis was possible. Common outcome measures included kidney weight, nephron number, glomerular size, glomerular filtration rate, and creatinine clearance. To date, there have been no studies assessing kidney function in large animal models. Most studies were rated as having a high or unknown risk of bias. The current body of evidence in animals suggests that exposure to maternal global nutrient restriction during pregnancy has detrimental effects on offspring kidney structure and function, such as lower kidney weight, lower nephron endowment, larger glomerular size, and lower glomerular filtration rate. Further long-term followup of studies in large animal models investigating kidney function through to adulthood are warranted.

Ruminant models of prenatal growth restriction

Intrauterine growth restriction (IUGR) is a significant health issue that not only affects infant mortality and morbidity, but may also predispose individuals to coronary heart disease, diabetes, hypertension and stroke as adults. The majority of IUGR pregnancies in humans are characterized by asymmetric fetal growth, resulting from inadequate nutrient transfer to the fetus. Furthermore, most of these pregnancies involve functional placental insufficiency, and may also show altered umbilical velocimetry. As the severity of IUGR increases, the fetus becomes increasingly hypoxic, hypoglycaemic and acidotic. In addition, placental transfer or utilization of some amino acids is known to be altered in IUGR pregnancies. Although a great deal has been learned from clinical studies of human IUGR, appropriate animal models are required to define completely the mechanisms involved in the development of IUGR. The pregnant sheep is a long-standing model for placental-fetal interactions, and fetal growth restriction can be induced in pregnant sheep by maternal nutrient restriction, maternal nutrient excess, administration of glucocorticoid, utero-placental embolization, carunclectomy and maternal hyperthermia. Although all of these sheep models are capable of inducing fetal growth restriction, the degree of restriction is variable. This review compares these sheep models of IUGR with the characteristics of human IUGR.

Handling has an anxiolytic effect that is not affected by the inhibition of the protein kinase C pathway in adult prenatal undernourished male rat offspring.

Environmental enrichment (EE) after birth has been reported as an intervention improving the anxiety-like behavior and cognitive deficit due to maternal restraint, foot-shock, or social stress during pregnancy. However, it remains unclear whether EE after birth could benefit the early prenatal undernourished offspring. In this study, we examined the effect of daily handling as a simple EE intervention on the aberrant behavior of prenatally undernourished rats. The male rat offspring exhibited anxiety-like behavior at 9 weeks of age due to maternal food restriction in early pregnancy. Our study shows that the daily handling after weaning has an anxiolytic effect in the prenatally undernourished offspring without affecting the behavior of prenatally well-nourished offspring. Conversely, the concentrations of dopamine, serotonin, norepinephrine, and their metabolites were not altered in the prefrontal cortex by prenatal undernutrition or daily handling after weaning. We investigated whether the anxiolytic effect of daily handling was mediated by the protein kinase C (PKC) pathway using the PKC inhibitor, chelerythrine. The anxiolytic effect of the handling was not canceled by chelerythrine injection in prenatally undernourished offspring, whereas chelerythrine induced an anxiety-like behavior in control rats. Our results suggest that maternal undernutrition in early pregnancy induces an anxiety-like behavior accompanied with a PKC pathway-hyporesponsiveness; however, daily handling ameliorates the anxiety-like behavior through a PKC-independent pathway.

Moderate nutrient restriction of beef heifers alters expression of genes associated with tissue metabolism, accretion, and function in fetal liver, muscle, and cerebrum by day 50 of gestation.

We hypothesized that a moderate maternal nutrient restriction during the first 50 d of gestation in beef heifers would affect transcript abundance of genes associated with tissue metabolism, accretion, and function in fetal liver, muscle, and cerebrum. Angus-cross heifers were estrus synchronized and assigned at breeding to one of two dietary treatments (CON- 100% of nutrient requirements to gain 0.45 kg/d; RES- 60% of CON). At day 50 of gestation, 14 heifers were ovariohysterectomized, and fetal liver, muscle, and cerebrum were collected. Transcriptome analysis via RNA-seq was conducted on the Illumina HiSeq 2500 platform using 50-bp paired-end reads at a depth of 2 × 10.4M reads/sample. Bioinformatic analysis was performed using the Tuxedo Suite and ontological analysis with DAVID 6.8. For fetal liver, muscle, and cerebrum, a total of 548, 317, and 151 genes, respectively (P < 0.01) were differentially expressed, of which 201, 144, and 28 genes, respectively were false discovery rate protected (FDR; q < 0.10). Differentially expressed genes were screened for fit into functional categories of pathways or ontologies associated with known impacts on tissue metabolism, accretion, and function. In fetal liver, five functional categories of interest (n = 125 genes) were affected by nutritional treatment: metabolic pathways, protein kinase, nucleosome core, mRNA splicing, and complement/coagulation cascades, of which 105 genes were upregulated in RES. In fetal muscle, three functional categories of interest (n = 106 genes) were affected by nutritional treatment: skeletal muscle, embryogenesis, and signaling cascades, of which 64 genes were upregulated in RES. In fetal cerebrum, three functional categories of interest (n = 60 genes) were affected by nutritional treatment: hippocampus and neurogenesis, metal-binding, and cytoskeleton, of which 58 genes were upregulated in RES. These results demonstrate that a moderate maternal nutrient restriction during the first 50 d of gestation in beef heifers alters transcript abundance of genes potentially impacting tissue metabolism, accretion, and function in fetal liver, muscle, and cerebrum. Furthermore, these results indicate that affected categories are tissue-specific and moderate maternal nutrient restriction generally increases expression of genes in fetuses from RES fed dams. Finally, these data lay the foundation upon which further research that identifies phenotypic responses to changes in these pathways may be elucidated.

Intergenerational response of steroidogenesis-related genes to maternal malnutrition.

We sought to examine whether rat maternal food restriction (MFR) affects the expression of steroidogenesis-related genes Cyp19, Cyp17a1, Insl3 and Gdf-9 in the ovaries of offspring from the first (FRG1) and second (FRG2) generations at pre-pubertal age (week 4) and during adulthood (week 8). At week 4, MFR significantly increased the expression of RNAs for all analyzed genes in both FRG1 and FRG2 females, which may indicate that MFR affects the onset of the reproductive lifespan, by inducing early pubertal onset. At week 8, the Cyp19 gene was still upregulated in MRF-subjected animals (Cyp19: P=0.0049 and P=0.0508 in FRG1 and FRG2, respectively), but MFR induced a significant decrease in Cyp17 and Gdf-9 gene expression in the offspring of both FRG1 and FRG2 females when compared with the controls (Cyp17: P=0.0018 and P=0.0016, respectively; Gdf-9: P=0.0047 and P=0.0023, respectively). This suggests that females at week 8, which should normally be in their optimal reproductive capacity, experience premature ovarian aging. At week 4, the activation of Cyp19 and Cyp17 was higher in the FRG1 ovaries than in the FRG2 ovaries, whereas the extent of Insl3 and Gdf-9 activation was lower in the FRG1 ovaries. This may indicate that FRG2 females were more vulnerable to MFR than their mothers (FRG1) and grandmothers, which is consistent with the 'predictive adaptive response' hypothesis. Our findings reveal that MFR may induce intergenerational ovarian changes as an adaptive response to ensure reproductive success before death.

Abstract # 2064 Hypercaloric diet in juvenile rats ameliorates sexual impairments and the increased TNF-alpha levels induced by maternal food restriction

The present study investigated whether a hypercaloric diet that is administered in rats during the juvenile period prevents sexual impairments that are caused by maternal food restriction and the tendency to become overweight/obese. Female rats were prenatally fed a 40% restricted diet from gestational day 2 to 18. The pups received a hypercaloric diet from postnatal day (PND) 23 to 65 (food restricted associated to hypercaloric [FRH] group) or laboratory chow (food restricted control [FRC] group). Pups from non-food-restricted dams received laboratory chow during the entire experiment (non-food-restricted [NFR] group). During the juvenile period and adulthood, body weight gain was evaluated weekly. Sexual behavior, hypodermal adiposity, and serum TNF-alpha levels were evaluated. Because obesity/overweight is related to inflammatory process the TNF-alpha study is important. The FRH group exhibited an increase in body weight on PND58 and 65. The FRC group exhibited an increase in the latency to the first mount and intromission, i.e., sexual behavior impairment. FRC also exhibited an increase in TNF-alpha levels. The hypercaloric diet reversed all of these behavioral and immune disturbances, but increased adiposity. We concluded that the hypercaloric diet administered during the juvenile period attenuated reproductive and immune disturbances that were induced by maternal food restriction, but did not reduce the tendency to become overweight/obese. The present results seem to be related to the thrifty phenotype hypothesis.

Offspring from maternal nutrient restriction in mice show variations in adult glucose metabolism similar to human fetal growth restriction.

Fetal growth restriction (FGR) is a pregnancy condition in which fetal growth is suboptimal for gestation, and this population is at increased risk for type 2 diabetes as adults. In humans, maternal malnutrition and placental insufficiency are the most common causes of FGR, and both result in fetal undernutrition. We hypothesized that maternal nutrient restriction (MNR) in mice will cause FGR and alter glucose metabolism in adult offspring. Pregnant CD-1 mice were subjected to MNR (70% of average ad libitum) or control (ad libitum) from E6.5 to birth. Following birth, mice were fostered by mothers on ad libitum feeds. Weight, blood glucose, glucose tolerance and tissue-specific insulin sensitivity were assessed in male offspring. MNR resulted in reduced fetal sizes but caught up to controls by 3 days postnatal age. As adults, glucose intolerance was detected in 19% of male MNR offspring. At 6 months, liver size was reduced (P = 0.01), but pAkt-to-Akt ratios in response to insulin were increased 2.5-fold relative to controls (P = 0.004). These data suggest that MNR causes FGR and long-term glucose intolerance in a population of male offspring similar to human populations. This mouse model can be used to investigate the impacts of FGR on tissues of importance in glucose metabolism.

The GLP-1 analog, liraglutide prevents the increase of proinflammatory mediators in the hippocampus of male rat pups submitted to maternal perinatal food restriction

BackgroundPerinatal maternal malnutrition is related to altered growth of tissues and organs. The nervous system development is very sensitive to environmental insults, being the hippocampus a vulnerable structure, in which altered number of neurons and granular cells has been observed. Moreover, glial cells are also affected, and increased expression of proinflammatory mediators has been observed. We studied the effect of Glucagon-like peptide-1 receptor (GLP-1R) agonists, liraglutide, which have very potent metabolic and neuroprotective effects, in order to ameliorate/prevent the glial alterations present in the hippocampus of the pups from mothers with food restriction during pregnancy and lactation (maternal perinatal food restriction—MPFR).MethodsPregnant Sprague-Dawley rats were randomly assigned to 50% food restriction (FR; n = 12) or ad libitum controls (CT, n = 12) groups at day of pregnancy 12 (GD12). From GD14 to parturition, pregnant FR and CT rats were treated with liraglutide (100 μg/kg) or vehicle. At postnatal day 21 and before weaning, 48 males and 45 females (CT and MPFR) were sacrificed. mRNA expression levels of interleukin-1β (IL1β), interleukin-6 (IL-6), nuclear factor-κβ, major histocompatibility complex-II (MHCII), interleukin 10 (IL10), arginase 1 (Arg1), and transforming growth factor (TGFβ) were assessed in the hippocampus by quantitative real-time polymerase chain reaction. Iba1 and GFAP-immunoreactivity were assessed by immunocytochemistry.ResultsThe mRNA expression IL1β, IL6, NF-κB, and MHCII increased in the hippocampus of male but not in female pups from MPFR. In addition, there was an increase in the percentage of GFAP and Iba1-immupositive cells in the dentate gyrus compared to controls, indicating an inflammatory response in the brain. On the other hand, liraglutide treatment prevented the neuroinflammatory process, promoting the production of anti-inflammatory molecules such as IL10, TGFβ, and arginase 1, and decreasing the number and reactivity of microglial cells and astrocytes in the hippocampus of male pups.ConclusionTherefore, the GLP-1 analog, liraglutide, emerges as neuroprotective drug that minimizes the harmful effects of maternal food restriction, decreasing neuroinflammation in the hippocampus in a very early stage.

Maternal nutrient restriction alters uterine artery hemodynamics and placentome vascular density in Bos indicus and Bos taurus.

The objective was to examine uterine artery blood flow (UBF) as well as macroscopic and microscopic placentome vascular density in nutrient-restricted Angus and Brahman heifers. Angus (n = 6) and Brahman (n = 6) heifers were bred to a single sire and pregnancy confirmed at 30-d postbreeding. Heifers were randomly assigned to 1 of 2 dietary treatments consisting of 100% (control-fed; CON; n = 6) or 60% (total nutrient-restricted; RES; n = 6) based from net energy requirements for gestating heifers. Nutritional treatments were imposed from days 50 to 180 of gestation. On day 175 of gestation, UBF was collected ipsilateral and contralateral to the conceptus via Doppler ultrasonography. Heifers underwent Cesarean sections for collection of 2 adjacent placentomes on day 180 of gestation. The primary cotyledonary artery of 1 placentome was perfused with Alexa Fluor 647 Con A conjugate to examine macroscopic cotyledonary vascular density via an in vivo imaging system. The second placentome was fixed for microscopic immunofluorescence labeling of capillaries and separated into maternal (caruncle) and fetal (cotyledon) components for determination of angiogenic factor mRNA expression. Main effects of nutritional treatment and breed are reported in the absence of a significant nutritional treatment by breed interaction. Ipsilateral UBF was decreased (P < 0.05) by 48% in RES vs. CON, whereas breed did not influence ipsilateral UBF. Contralateral UBF was not different between nutritional treatments; however, contralateral UBF was decreased (P < 0.05) by 63% in Brahman vs. Angus cattle. Macroscopic cotyledonary vascular density was increased (P < 0.05) by 36% in RES vs. CON and 82% in Brahman vs. Angus heifers. Percent capillary area and capillary perimeter were increased (P < 0.05) in RES vs. CON and increased (P < 0.05) in Brahman vs. Angus heifers. Dietary treatments did not alter angiogenic factor expression; however, transcript abundance of caruncle and cotyledon ANGP1, FLT1, and KDR was increased (P < 0.05) in Brahman vs. Angus heifers. In summary, these data indicate compensatory responses in macroscopic and microscopic placentome blood vessel density during maternal nutrient restriction-induced reductions in UBF. Moreover, a greater macroscopic density of cotyledonary blood vessels was observed in Brahman vs. Angus heifers.

Influences of maternal nutrient restriction and arginine supplementation on visceral metabolism and hypothalamic circuitry of offspring

Maternal nutrient restriction during gestation can exert long-term negative effects on offspring health and performance. Arginine supplementation may rescue some of the negative effects elicited by maternal nutrient restriction. We tested the hypothesis that maternal arginine supplementation during gestation would rescue deleterious effects of nutrient restriction on in vitro O2 consumption in the liver and jejunum and hypothalamic protein expression of proopiomelanocortin (POMC), neuropeptide Y (NPY), agouti-related peptide (AgRP), and neuronal nitric oxide synthase (nNOS), and the colocalization of nNOS and active phosphor-signal transducer and activator of transcription 3 (pSTAT3) in female offspring. Multiparous ewes were assigned to dietary treatment at 54 d of gestation: 100% of requirements (Con), 60% of control (Res), or Res plus rumen-protected arginine (Res-Arg; 180 mg/kg). At parturition, offspring were immediately removed from their dam and placed on a common diet. At 54 ± 4 d of age, female lambs (n = 6 per treatment) were weighed, the liver and jejunum were weighed, and samples were collected for in vitro measurement of O2 consumption. The hypothalamus was collected to determine protein expression of POMC, NPY, AgRP, and nNOS, and the colocalization of nNOS and pSTAT3 (n = 3, 4, and 4 for Con, Res, and Res-Arg, respectively). Hepatic consumption of O2 in vitro (mol/min/liver) was decreased (P = 0.04) in the Res and Res-Arg group compared with Con. Intensity of staining for NPY-containing fibers tended to decrease (P = 0.10) in Res and Res-Arg compared with Con. Number of POMC neuronal cells in the arcuate nucleus (ARC) of the hypothalamus decreased (P ≤ 0.03) in the Res group compared with Res-Arg. These observations demonstrate that maternal nutrient restriction decreases energy utilization in the liver and number of POMC cells in the ARC of offspring. Supplementation of arginine to the gestating ewe failed to influence hepatic use of energy in lambs from Res ewes. Numbers of POMC-containing cells were increased in the ARC in lambs from ewes restricted to 60% of nutritional requirements and supplemented with rumen-protected arginine, potentially influencing feeding behavior and hepatic energy metabolism.

Perinatal and long-term effects of maternal uterine artery adenoviral VEGF-A165 gene therapy in the growth-restricted guinea pig fetus.

Uterine artery application of adenoviral vascular endothelial growth factor A165 (Ad.VEGF-A165) gene therapy increases uterine blood flow and fetal growth in experimental animals with fetal growth restriction (FGR). Whether Ad.VEGF-A165 reduces lifelong cardiovascular disease risk imposed by FGR remains unknown. Here, pregnant guinea pigs fed 70% normal food intake to induce FGR received Ad.VEGF-A165 (1×1010 viral particles, n = 15) or vehicle ( n = 10), delivered to the external surface of the uterine arteries, in midpregnancy. Ad libitum-fed controls received vehicle only ( n = 14). Litter size, gestation length, and perinatal mortality were similar in control, untreated FGR, and FGR+Ad.VEGF-A165 animals. When compared with controls, birth weight was lower in male but higher in female pups following maternal nutrient restriction, whereas both male and female FGR+Ad.VEGF-A165 pups were heavier than untreated FGR pups ( P < 0.05, ANOVA). Postnatal weight gain was 10-20% greater in female FGR+Ad.VEGF-A165 than in untreated FGR pups, depending on age, although neither group differed from controls. Maternal nutrient restriction reduced heart weight in adult female offspring irrespective of Ad.VEGF-A165 treatment but did not alter ventricular wall thickness. In males, postnatal weight gain and heart morphology were not affected by maternal treatment. Neither systolic, diastolic, mean arterial pressure, adrenal weight, nor basal or challenged plasma cortisol were affected by maternal undernutrition or Ad.VEGF-A165 in either sex. Therefore, increased fetal growth conferred by maternal uterine artery Ad.VEGF-A165 is sustained postnatally in FGR female guinea pigs. In this study, we did not find evidence for an effect of maternal nutrient restriction or Ad.VEGF-A165 therapy on adult offspring blood pressure.