Maternal Nutrient Restriction Research Articles

Combined Growth Hormone (GH) and Insulin-Like Growth Factor-I (IGF-I) Treatment is More Effective than GH or IGF-I Alone at Enhancing Recovery from Neonatal Malnutrition in Rats

The effect of hormonal therapy on enhancing recovery from neonatal malnutrition was assessed in rats. Malnutrition was induced by maternal food restriction (60% control intake) during lactation. On d 16 postpartum, restricted pups were refed by cross-fostering to control dams and were subcutaneously administered growth hormone (GH), insulin-like growth factor-I (IGF-I), GH + IGF-I or saline. At d 21, pups were weaned and continued hormonal treatments until d 39 postpartum. By d 39, body weight of GH (96% control) and GH + IGF-I- (98%) treated animals were normal, whereas IGF-I (88%) and saline- (85%) treated animals were still stunted. Hormone effectiveness was age dependent, with growth rates of GH + IGF-I-, IGF-I- and GH-treated pups being greatest between d 16 and 30, d 16 and 21 and after d 30, respectively. Protein accretion by d 39 was higher (P < 0.01) in GH and GH + IGF-I groups than saline- or IGF-I-treated groups. On d 39, serum IGF-I concentrations were as follows: GH + IGF-I > IGF-I > GH = saline (placebo), indicating that an elevated serum IGF-I was not required for GH-stimulated growth. Of the three hormonal treatments, GH + IGF-I therapy was most efficacious at promoting rapid and complete body weight recovery and supporting lean body mass accretion.

Human Growth Hormone but Not Human Insulin-Like Growth Factor-I Enhances Recovery from Neonatal Malnutrition in Rats

The ability of hormonal therapy to enhance recovery from neonatal mainutrition was assessed in rats. Pups were mainourished via maternal food restriction (60% control intake). On d 16, restricted pups (n = 50) (62% control body wt) were refed until d 20 and were given growth hormone, insulin-like growth factor-I, destripeptide [1,3] insulin-like growth factor-I, or saline (placebo). Refed-placebo pups attained 80% of control body weight by d 20. Growth hormone treatment caused a greater weight gain than refeeding alone (P ≤ 0.05). Treatment with insulin-like growth factor-I and destripeptide [1,3] insulin-like growth factor-I did not affact body weight. All three hormones increased spleen and kidney weights (P ≤ 0.05) compared with the refedplacebo group, whereas only growth hormone increased muscle weight (P ≤0.05). Mainourished pups had lower serum insulin-like growth factor-I (P ≤ 0.05) and insulin-like growth factor binding protein-3 (P ≤ 0.05), and higher serum insulin-like growth factor binding protein-2 (P ≤ 0.05) and hepatic insulin-like growth factor binding protein-2 mRNA (P ≤ 0.05) than controls. Refeeding increased serum insulin-like growth factor-I compared with restricted pups (P ≤ 0.05), and insulin-like growth factor-I treatment caused a further increased in serum insulin-like growth factor-I compared with the refedplacebo group (P ≤ 0.05). These results show that growth hormone was moderately effective at increasing body weight gain and muscle growth during recovery from mainutrition.

Placental nanoparticle gene therapy normalizes gene expression changes in the fetal liver associated with fetal growth restriction in a fetal sex-specific manner.

Fetal growth restriction (FGR) is associated with increased risk of developing non-communicable diseases. We have a placenta-specific nanoparticle gene therapy protocol that increases placental expression of human insulin-like growth factor 1 (hIGF1), for the treatment of FGR in utero. We aimed to characterize the effects of FGR on hepatic gluconeogenesis pathways during early stages of FGR establishment, and determine whether placental nanoparticle-mediated hIGF1 therapy treatmentcould resolve differences in the FGR fetus. Female Hartley guinea pigs (dams) were fed either a Control or Maternal Nutrient Restriction (MNR) diet using established protocols. At GD30-33, dams underwent ultrasound guided, transcutaneous, intraplacental injection of hIGF1 nanoparticle or PBS (sham) and were sacrificed 5 days post-injection. Fetal liver tissue was fixed and snap frozen for morphology and gene expression analysis. In female and male fetuses, liver weight as a percentage of body weight was reduced by MNR, and not changed with hIGF1 nanoparticle treatment. In female fetal livers, expression of hypoxia inducible factor 1 (Hif1α) and tumor necrosis factor (Tnfα) were increased in MNR compared to Control, but reduced in MNR + hIGF1 compared to MNR. In male fetal liver, MNR increased expression of Igf1 and decreased expression of Igf2 compared to Control. Igf1 and Igf2 expression was restored to Control levels in the MNR + hIGF1 group. This data provides further insight into the sex-specific mechanistic adaptations seen in FGR fetuses and demonstrates that disruption to fetal developmental mechanisms may be returned to normal by treatment of the placenta.

Decreased ubiquitin modifying enzyme A20 associated with hyper-responsiveness to ovalbumin challenge following intrauterine growth restriction

BackgroundIntrauterine growth restriction (IUGR) is strongly correlated with an increased risk of asthma later in life. Farm dust protects mice from developing house dust mite-induced asthma, and loss of ubiquitin modifying enzyme A20 in lung epithelium would abolish this protective effect. However, the mechanisms of A20 in the development of asthma following IUGR remains unknown.MethodsAn IUGR rat model induced by maternal nutrient restriction was used for investigating the role of A20 in the response characteristics of IUGR rats to ovalbumin (OVA) challenge. The ubiquitination of proteins and N6-methyladenosine (m6A) modifications were used to further assess the potential mechanism of A20.ResultsIUGR can reduce the expression of A20 protein in lung tissue of newborn rats and continue until 10 weeks after birth. OVA challenging can increase the expression of A20 protein in lung tissue of IUGR rats, but its level was still significantly lower than the control OVA group. The differentially ubiquitinated proteins in lung tissues were also observed in IUGR and normal newborn rats. Furthermore, this ubiquitination phenomenon continued from the newborn to adulthood. In the detected RNA methylations, m6A abundance of the motif GGACA was the highest. The higher abundances of m6A modification of A20 mRNA from IUGR were negatively correlated with the trend of A20 protein levels.ConclusionThese findings indicate A20 as a key regulator during the development of asthma following IUGR, providing further insight into the prevention of asthma induced by environmental factors.

Placental treatment with insulin-like growth factor 1 via nanoparticle differentially impacts vascular remodeling factors in guinea pig sub-placenta/decidua.

Clinically, fetal growth restriction (FGR) is only detectable in later gestation, despite pathophysiological establishment likely earlier in pregnancy. Additionally, there are no effective in utero treatment options for FGR. We have developed a nanoparticle to deliver human insulin-like 1 growth factor (hIGF-1) in a trophoblast-specific manner which results in increased expression of hIGF-1. IGF-1 signaling in the placenta regulates multiple developmental processes including trophoblast invasion and maternal vascular remodeling, both of which can be diminished in the FGR placenta. We aimed to determine the effects of short-term hIGF-1 nanoparticle treatment on sub-placenta/decidua trophoblast signaling mechanisms in FGR and under normal growth conditions. Using the guinea pig maternal nutrient restriction (MNR) model of FGR, ultrasound-guided, intra-placenta injections of hIGF-1 nanoparticle were performed at gestational day 30-33, and dams sacrificed 5days later. Sub-placenta/decidua tissue was separated from placenta for further analyses. Western blot was used to analyze protein expression of ERK/AKT/mTOR signaling proteins (phospho-Erk (pERK), phospho-Akt (pAKT), raptor, rictor and deptor). qPCR was used to analyze gene expression of vascular/remodeling factors [vascular endothelial growth factor (Vegf), placenta growth factor (Pgf), platelet-derived growth factor (Pdgf)) and tight junction/adhesion proteins (claudin 5 (Cldn5), p-glycoprotein (Abcb1), occludin (Ocln) and tight junction protein 1 (Zo1)]. MNR reduced expression of pERK, PdgfB and Cldn5, and increased expression of Ocln and Zo1 in the sub-placenta/decidua. In MNR + hIGF1 nanoparticle sub-placenta/decidua, expression of PdgfB, Ocln and Zo1 was normalized, whilst pAkt, VegfB, Vegf receptor 1 and PdgfB receptor were increased compared to MNR. In contrast, hIGF-1 nanoparticle treatment of normal placentas reduced expression of pERK, raptor and increased expression of the mTOR inhibitor deptor. This was associated with reduced expression of VegfA, Plgf, and PdgfB. Here we have shown that the impact of hIGF-1 nanoparticle treatment is dependent on pregnancy environment. Under MNR/FGR, hIGF-1 nanoparticle treatment triggers increased expression of growth factors and normalization of EMT factors. However, under normal conditions, the response of the placenta is to decrease AKT/mTOR signaling and growth factor expression to achieve homeostasis.

Timing of maternal nutrient restriction during mid- to late-gestation influences net umbilical uptake of glucose and amino acids in adolescent sheep.

Previous research demonstrated that maternal nutrient restriction during mid- to late-gestation influenced net umbilical uptakes of glucose and amino acids in sheep. However, it is unclear how the timing and duration of nutrient restriction during mid- to late-gestation influences net uterine, uteroplacental, and fetal flux of glucose and amino acids. On day 50 of gestation, 41 adolescent ewe lambs carrying singletons were randomly assigned to one of six dietary treatments: 1) 100% of nutrient requirements from days 50 to 90 of gestation (CON; n = 7); 2) 60% of nutrient requirements (RES; n = 7) from days 50 to 90 of gestation; 3) 100% of nutrient requirements from days 50 to 130 of gestation (CON-CON; n = 6); 4) 100% of nutrient requirements from days 50 to 90 of gestation and 60% of nutrient requirements from days 90 to 130 of gestation (CON-RES; n = 7); 5) 60% of nutrient requirements from days 50 to 90 of gestation and 100% of nutrient requirements from days 90 to 130 of gestation (RES-CON; n = 7); or 6) 60% of nutrient requirements from days 50 to 130 of gestation (RES-RES; n = 7). On day 90 (n = 14) and day 130 (n = 27), intraoperative procedures were performed to evaluate uteroplacental blood flows, collect blood samples, and then ewes were euthanized. Net uterine, uteroplacental, and umbilical fluxes of glucose and amino acids were calculated by multiplying blood flow by the arterial-venous concentration difference. Data from days 90 and 130 were analyzed separately using ANOVA in SAS. Maternal nutrient restriction during mid-gestation increased (P = 0.04) net umbilical glucose uptake but, maternal nutrient restriction during late-gestation decreased (P = 0.02) net umbilical glucose uptake. Net umbilical essential amino acid uptake decreased (P = 0.03) with nutrient restriction during mid-gestation; however, net umbilical uptakes of Phe (P = 0.02), Thr (P = 0.05), Met (P = 0.09), and His (P = 0.08) increased or tended to increase after nutrient restriction during late-gestation. These data demonstrate that net umbilical glucose and amino acid uptakes were influenced by the timing of nutrient restriction during mid- to late-gestation. Elevated net umbilical glucose uptake after mid-gestational nutrient restriction was sustained throughout late-gestation, independent of late-gestational feeding level. Long-term adaptations in umbilical glucose uptake may have implications for prenatal and postnatal growth and development of the offspring.

Maternal nutrient restriction during late gestation reduces vigor and alters blood chemistry and hematology in neonatal beef calves.

Fall-calving primiparous beef females [body weight: 451 ± 28 (SD) kg; body condition score: 5.4 ± 0.7] were individually-fed either 100% (control; CON; n = 13) or 70% (nutrient restricted; NR; n = 13) of metabolizable energy and metabolizable protein requirements for maintenance, pregnancy, and growth from day 160 of gestation to parturition. Calves were reared naturally by their dams and monitored for latency times from birth to first sternal recumbency, attempt to stand, and stand; vigor scores were assigned at 2, 5, 10, and 20min of age. Rectal temperatures and jugular blood were obtained at 0 (pre-suckling), 6, 12, 24, and 48h of age, and blood chemistry, hematology, cortisol, and insulin were determined. Data were analyzed with fixed effects of late gestational nutritional plane (single data point) or nutritional plane, hour, and their interaction (data over time, repeated measures). Calving date was a fixed effect; calf sex was included when P < 0.25. We previously reported that late gestational nutritional plane did not affect gestation length or calf size at birth, but calving assistance and fetal malpresentations occurred more often in NR. Nutritional plane did not affect (P = 0.65) duration of parturition, but calves born to NR dams had slower times to attempt to stand (P = 0.09), slower times to stand (P = 0.02), and had poorer 20min vigor scores (P = 0.05). Serum immunoglobulin G and A concentrations at 48h were greater (P ≤ 0.03) for NR calves. Rectal temperature of NR calves was less (P = 0.02) at 0h, but greater (P = 0.04) at 24h compared with CON. Circulating glucose, non-esterified fatty acids, triglycerides, cortisol, and insulin were not affected by nutritional plane (P ≥ 0.18). Total protein and globulin from 6 to 48h were greater (P ≤ 0.02) in NR calves. Calves from NR dams had greater (P < 0.08) gamma-glutamyl transferase at 6, 12, and 48h. Serum aspartate aminotransferase was greater (P ≤ 0.07) from 0 to 24h and creatine kinase was greater (P ≤ 0.04) from 6 to 24h in NR calves. At 0h, potassium was greater (P = 0.03) in NR calves. Calves born to CON had greater chloride (P = 0.08; main effect), sodium (P ≤ 0.09) from 0 to 48h, and anion gap (P = 0.02) at 6h. Hematocrit from 6 to 24h and red blood cells and hemoglobin at 6 and 12h were greater (P ≤ 0.09) in CON calves. These data indicate that nutrient restriction during late gestation resulted in less vigorous calves with more indicators of trauma in early life.

Maternal nutrient restriction and dietary melatonin alter neurotransmitter pathways in placental and fetal tissues

IntroductionRecent research indicates an important role in the placental fetal brain axis, with a paucity of information reported in large animals. Melatonin supplementation has been investigated as a potential therapeutic to negate fetal growth restriction. We hypothesized that maternal nutrient restriction and melatonin supplementation would alter neurotransmitter pathways in fetal blood, cotyledonary and hypothalamus tissue. MethodsOn day 160 of gestation, Brangus heifers (n = 29 in fall study; n = 25 in summer study) were assigned to one of four treatments: adequately fed (ADQ-CON; 100% NRC recommendation), nutrient restricted (RES-CON; 60% NRC recommendation), and ADQ or RES supplemented with 20 mg/d of melatonin (ADQ-MEL; RES-MEL). Placentomes, fetal blood, and hypothalamic tissue were collected at day 240 of gestation. Neurotransmitters were analyzed in fetal blood and fetal and placental tissues. Transcript abundance of genes in the serotonin pathway and catecholamine pathway were determined in fetal hypothalamus and placental cotyledon. ResultsSerotonin was increased (P < 0.05) by 12.5-fold in the blood of fetuses from RES dams versus ADQ in the fall study. Additionally, melatonin supplementation increased (P < 0.05) neurotransmitter metabolites and transcript abundance of the monoamine oxidase A (MAOA) enzyme in the cotyledon. In the summer study, plasma dopamine and placental dopamine receptors were decreased (P < 0.05) in RES dams versus ADQ. DiscussionIn conclusion, these data indicate novel evidence of the presence of neurotransmitters and their synthesis and metabolism in the bovine conceptus, which could have greater implications in establishing postnatal behavior.

PSII-9 Preliminary Investigation of Nutrient Challenge During Pregnancy on Wether Lamb Development, Growth, and Performance from Birth to Harvest

Abstract Inadequate nutrition during gestation has been shown to impair growth and metabolism of affected offspring and can predispose them to lifelong inefficiency and poor performance. Development throughout preweaning has been studied and compared to carcass performance; however, the impact of nutrient challenge during gestation on post weaning growth, efficiency, and nutrition utilization is not well known. Thus, our objective was to investigate the impacts of maternal nutrient restriction in late gestation on wether lambs' lifelong performance, from birth through harvest. We hypothesized that lambs from nutrient restricted dams would have reduced growth efficiency, taking longer to reach market weight and yielding altered body and carcass composition. Twenty-two bred yearling ewes were fed a diet meeting 100% of National Research Council (NRC) requirements until the 110th day of gestion (dGA). From 110 dGA until lambing occurred, ewes were either maintained on a diet meeting 100% of NRC requirements (n = 10) or reduced to a diet of 65% of NRC requirements (n = 12). Beginning at birth, body weight and morphometrics were collected weekly for control (CONT; n = 4) and treatment (NR; n = 5) wether lambs. At approximately 71 days of age, wethers were weaned and placed into individual pens to simulate a controlled feedlot setting. Wether lambs were fed a commercial lamb finishing diet ad libitum and orts were collected to evaluate feed efficiency and growth. At 115 days of age, wethers were placed in metabolism crates for 5 days to allow for total collection or urine and feces in order to evaluate nutrient balance. Birthweight, weaning weight, and morphometrics did not differ between NR and CONT lambs from birth until weaning. However, there was a tendency (P &amp;lt; 0.10) for a slower rate of gain from birth until weaning in NR lambs compared to their CONT counterparts. Current research in feedlot performance and digestibility is ongoing; however, thus far there are no differences in feed intake or average daily gain. These data suggest that a moderate nutrient challenge in the last month of gestation is not substantial enough to induce adaptive changes that result in differences in postnatal growth and performance.

395 Young Scholar Award Talk: Melatonin: A Promising Therapeutic for Compromised Pregnancies

Abstract Poorly functioning placenta restricts oxygen and nutrient supply to the fetus resulting in fetal nutrient deficiency and intrauterine growth restriction. Intrauterine growth restriction dramatically increases neonatal mortality and those who survive are more prone to experience metabolic diseases such as cardiovascular diseases, hypertension, and type 2 diabetes in later life. Therefore, the placenta plays a critical role in the developmental origins of health and diseases. Currently, there is a vast effort evaluating the effects of compromised pregnancies such as maternal undernutrition, overnutrition, and heat stress on fetal development; however fewer studies have evaluated potential therapeutics to enhance placental efficiency and offspring performance. Melatonin, a hormone that modulates circadian rhythms, has shown to increase umbilical and uterine blood flow in pregnant dams. Moreover, a recent study observed that melatonin therapeutic effects during compromised pregnancies are seasonally dependent. Dietary melatonin supplementation to pregnant beef cattle increased uterine blood flow, rescued fetal weights, increased angiogenic factors in the placenta, and reduced vaginal temperatures in the summer, but not in the fall. The same study found that maternal nutrient restriction was more detrimental during the fall reducing uterine blood flow. Additionally, it was observed that dams that are nutrient restricted during the summer exerted a compensatory mechanism at the placentome level where they exhibited greater placentome vascularity and increased placental capillary size and angiogenic factors. Lastly, bovine placental circadian rhythms have been identified and interestingly, temporal increases in placental vascularity occur concomitantly with greater melatonin receptor 1A transcript abundance. In conclusion, these findings provide a better understanding of placental insufficiency and impaired fetal growth during maternal nutrient restriction, while temporal alterations in the placenta could lead to the development of proper guidelines for administration of placental blood flow therapeutics such as melatonin. Future studies should evaluate melatonin impacts on offspring organ development and potential for mitigating heat stress in livestock.

298 Late Gestational Nutrient Restriction of Primiparous Beef Heifers Decreases Neonatal Calf Body Weight and Organ Mass

Abstract We hypothesized that late gestational nutrient restriction (NR) would decrease organ mass in neonatal calves. Fall-calving crossbred beef heifers (BW: 466 ± 30 [SD] kg; BCS: 5.4 ± 0.5) bred to a single sire were individually fed 100% (control; n = 10) or 70% (nutrient restricted; NR; n = 12) of NASEM metabolizable energy and metabolizable protein requirements for maintenance, pregnancy, and growth from d 160 of gestation to calving. Diets were based on chopped sorghum sudan hay and supplemented to meet targeted nutritional planes. Calves were removed immediately post-calving and fed colostrum and milk replacer relative to BW until euthanasia (48.7 ± 0.4 h of age) for organ mass determination. Nutritional plane was a fixed effect, and maternal sire, calf sex (when P &amp;lt; 0.25), and date of study initiation were covariates. Post-calving, NR dams were 62 kg BW and 2.1 BCS less (P &amp;lt; 0.001) than controls. Calf BW, liver, large intestine, omental/mesenteric fat, kidney, and heart weights were less (P ≤ 0.05) and carcass, rumen/reticulum/omasum, stomach complex, and total gastrointestinal tract weights tended to be less (P ≤ 0.09) in offspring from NR dams. Maternal nutrient restriction did not affect (P ≥ 0.11) weights of the small intestine, pancreas, abomasum, spleen, perirenal fat, adrenals, lungs, femur, and brain. Expressed per BW, lung weight was greater (P = 0.05), and brain and femur weight tended to be greater (P ≤ 0.10) in calves from NR dams, but other organ masses were unaffected (P ≥ 0.15). Expressed relative to brain weight, liver and large intestine weights were less (P ≤ 0.04) and carcass, total gastrointestinal tract, omental/mesenteric fat, and kidney weights tended to be less (P ≤ 0.08) in NR. In summary, late gestational NR decreased some organ masses in neonatal calves while sparing other tissues via asymmetric growth.

210 Restricted Maternal Nutrition: Impacts on Fetal Development and Metabolism in Sheep

Abstract Adequate nutrition is important for proper growth and metabolism. This is even more important during gestation when insults to the fetus can negatively impact fetal growth and development with persistent effects into adulthood. Using a sheep model, we have demonstrated that maternal restricted feeding (60% of National Research Council requirements) during gestation reduced growth in offspring and muscle fiber cross-sectional area. These changes in growth are associated with reduced stem cell (satellite cells and mesenchymal stem cells) population and function. Offspring of restricted-fed mothers also displayed altered metabolism indicated by increased insulin resistance, leptin dysregulation, and reduced circulating metabolic and growth factors. Based on these persistent negative effects of maternal diet on offspring postnatal growth and metabolism, we evaluated the impacts during fetal growth on whole body and tissue-specific metabolism and the impact of maternal realimentation during gestation. Nutrient restriction caused changes in metabolic pathways in a tissue specific manner in muscle, liver, and blood demonstrating the ability of maternal diet to program offspring metabolism in highly metabolic tissues. Several pathways involved in amino acid, lipid, carbohydrate, and transsulfuration/methionine metabolism were altered in all 3 tissues, demonstrating the whole-body effects of maternal diet on offspring metabolism. Interestingly, realimentation alleviated some of the effects of nutrient restriction on offspring metabolic pathways. In summary, maternal nutrient restriction during gestation negatively affects offspring fetal growth with persistent effects into adulthood which may be caused by changes in stem cell function and key metabolic pathways during fetal development.

97 Maternal Nutrient Restriction and Melatonin Supplementation Alter Neurotransmitter Profiles in Bovine Fetal and Placental Tissues

Abstract The placenta modulates fetal brain development in mice and humans, with a paucity of information reported in cattle. The objective was to determine the effects of maternal nutrient restriction and melatonin supplementation on neurotransmitter concentrations in the cotyledon, fetal hypothalamus, and fetal blood. At day 160 of gestation, 29 spring-calving and 25 fall-calving Brangus heifers were assigned treatments: adequately fed (ADQ-CON; 100% NRC recommendation), nutrient restricted (RES-CON; 60% NRC recommendation), and ADQ or RES supplemented with 20 mg/d of melatonin (ADQ-MEL; RES-MEL). Fetuses were removed at day 240 of gestation for collection of placentomes, blood, and hypothalamic tissue. Neurotransmitters (serotonin, 5-HT; dopamine, DA; norepinephrine, NE; epinephrine, EPI) and metabolites (3,4-dihydroxyphenylacetic acid, DOPAC; 5-hydroxyindoleacetic acid, 5-HIAA; homovanillic acid HVA) were analyzed by LCMS or HPLC in blood or tissues, respectively. Data were analyzed by ANOVA using the mixed procedure in SAS with seasons analyzed separately. In spring-calving heifers, melatonin increased (P ≤ 0.05) concentrations of DOPAC, DA, 5-HIAA, and HVA in the cotyledon. Nutrient restriction and melatonin supplementation increased (P ≤ 0.05) blood 5-HT. In both calving seasons, there was a nutrition by treatment interaction in hypothalamic 5-HT (P ≤ 0.01), whereby RES-MEL was increased compared with ADQ-MEL. For fall-calving heifers, no differences were observed in neurotransmitter concentrations in the cotyledon. Hypothalamic NE was increased (P = 0.02) in RES vs. ADQ, while hypothalamic and blood DA were decreased (P ≤ 0.05) in RES vs. ADQ. Hypothalamic 5-HIAA was increased (P = 0.05) in the MEL vs. CON, while blood EPI was decreased (P = 0.04) in MEL vs. CON. In summary, nutrient restriction and melatonin supplementation alter concentrations of neurotransmitters and their metabolites in a seasonally dependent manner. Further investigation into the gene expression of these neurotransmitter pathways is necessary to better define the bovine placental-brain axis.

Nanoparticle-mediated transgene expression of insulin-like growth factor 1 in the growth restricted guinea pig placenta increases placenta nutrient transporter expression and fetal glucose concentrations.

Fetal growth restriction (FGR) significantly contributes to neonatal and perinatal morbidity and mortality. Currently, there are no effective treatment options for FGR during pregnancy. We have developed a nanoparticle gene therapy targeting the placenta to increase expression of human insulin-like growth factor 1 (hIGF1) to correct fetal growth trajectories. Using the maternal nutrient restriction guinea pig model of FGR, an ultrasound-guided, intraplacental injection of nonviral, polymer-based hIGF1 nanoparticle containing plasmid with the hIGF1 gene and placenta-specific Cyp19a1 promotor was administered at mid-pregnancy. Sustained hIGF1 expression was confirmed in the placenta 5 days after treatment. Whilst increased hIGF1 did not change fetal weight, circulating fetal glucose concentration were 33%-67% higher. This was associated with increased expression of glucose and amino acid transporters in the placenta. Additionally, hIGF1 nanoparticle treatment increased the fetal capillary volume density in the placenta, and reduced interhaemal distance between maternal and fetal circulation. Overall, our findings, that trophoblast-specific increased expression of hIGF1 results in changes to glucose transporter expression and increases fetal glucose concentrations within a short time period, highlights the translational potential this treatment could have in correcting impaired placental nutrient transport in human pregnancies complicated by FGR.

Effects of nutrient restriction and subsequent realimentation in pregnant beef cows: Maternal endocrine profile, umbilical hemodynamics, and mammary gland development and hemodynamics

Our hypothesis was that maternal nutrient restriction would negatively impact the endocrine and metabolic status of the pregnant cow, therefore influencing the mammary gland in preparation for lactation. We further hypothesized that earlier timing of realimentation could prevent negative impacts of nutrient restriction. The objectives were to investigate the influence of nutrient restriction and realimentation during early to late gestation on endocrine profile, umbilical hemodynamics, and mammary gland development and hemodynamics in pregnant beef cows. In Experiment 1, on d 30 of pregnancy cows (initial BW = 667.5 ± 13.4 kg, BCS = 6.2 ± 0.1) were randomly assigned to one of 3 treatments: 1) 100% NRC requirements from d 30 to 254 of gestation (CCC; n = 6); 2) 60% NRC from d 30 to 85, thereafter being re-alimented to 100% NRC to d 254 (RCC; n = 5); 3) or receive 60% NRC from d 30 to 140, thereafter being re-alimented to 100% NRC to d 254 (RRC; n = 6). Cows were returned to a common outdoor facility for calving thereafter and were fed ad libitum. In Experiment 2, on d 30 of pregnancy, cows (initial BW = 620.5 ± 11.3 kg, BCS = 5.1 ± 0.1) were randomly assigned to dietary treatments including: control (CON; 100% NRC; n = 18) and nutrient restriction (RES; 60% NRC; n = 30). On d 85 of pregnancy, cows were either slaughtered (CON, n = 6 and RES, n = 6), remained on control (CC; n = 12) and restricted (RR; n = 12) treatments, or were realimented to control (RC; n = 11). On d 140 of pregnancy, cows were either slaughtered (CC, n = 6; RR, n = 6; RC, n = 5), remained on control (CCC, n = 6; RCC, n = 5), or were realimented to control (RRC, n = 6). On d 254 of pregnancy, all remaining cows were slaughtered (CCC, n = 6; RCC, n = 5; RRC, n = 6). Mammary hemodynamics and endocrine profile were measured. Serum urea nitrogen, NEFA, as well as fetal parameters were measured in Experiment 1; whereas in Experiment 2, mammary gland development was recorded. In Experiment 1, RRC cows had lower dry matter intake (P = 0.001) and consequently lower BW change (P = 0.06). However, maternal nutrition did not alter mammary hemodynamics, hormonal patterns, and fetal characteristics (P > 0.11). In Experiment 2, CCC cows had increased (P = 0.02) mammary gland blood flow ipsilateral to the gravid horn as well as greater (P = 0.02) mammary gland fat on d 254. Nevertheless, plane of nutrition did not alter hormonal concentrations nor mammary gland characteristics (P > 0.15). These data indicate that nutrient restriction did not alter mammary hemodynamics nor endocrine profile throughout gestation.

O23 Maternal nutrient restriction in beef heifers during early gestation increases B12 and folate concentrations in fetal allantoic fluids

O23 Maternal nutrient restriction in beef heifers during early gestation increases B12 and folate concentrations in fetal allantoic fluids

Effects of Maternal Nutrient Restriction and Melatonin Supplementation on Cardiomyocyte Cell Development Parameters Using Machine Learning Techniques.

Simple SummaryThe major objectives of this study were to examine the effects of maternal feed restriction and melatonin supplementation on fetal cardiomyocyte cell development parameters and predict binucleation and hypertrophy using machine learning techniques using pregnant beef heifers. Interestingly, major findings include evidence that compromised pregnancy in cattle leads to a reduction in the number of cardiomyocytes while melatonin treatment can mitigate some of these disturbances.The objective of the current study was to examine the effects of maternal feed restriction and melatonin supplementation on fetal cardiomyocyte cell development parameters and predict binucleation and hypertrophy using machine learning techniques using pregnant beef heifers. Brangus heifers (n = 29) were assigned to one of four treatment groups in a 2 × 2 factorial design at day 160 of gestation: (1) 100% of nutrient requirements (adequately fed; ADQ) with no dietary melatonin (CON); (2) 100% of nutrient requirements (ADQ) with 20 mg/d of dietary melatonin (MEL); (3) 60% of nutrient requirements (nutrient-restricted; RES) with no dietary melatonin (CON); (4) 60% of nutrient requirements (RES) with 20 mg/d of dietary melatonin (MEL). On day 240 of gestation, fetuses were removed, and fetal heart weight and thickness were determined. The large blood vessel perimeter was increased in fetuses from RES compared with ADQ (p = 0.05). The total number of capillaries per tissue area exhibited a nutrition by treatment interaction (p = 0.01) where RES-MEL increased capillary number compared (p = 0.03) with ADQ-MEL. The binucleated cell number per tissue area showed a nutrition by treatment interaction (p = 0.010), where it was decreased in RES-CON vs. ADQ-CON fetuses. Hypertrophy was estimated by dividing ventricle thickness by heart weight. Based on machine learning results, for the binucleation and hypertrophy target variables, the Bagging model with 5 Decision Tree estimators and 3 Decision Tree estimators produced the best results without overfitting. In the prediction of binucleation, left heart ventricular thickness feature had the highest Gin importance weight followed by fetal body weight. In the case of hypertrophy, heart weight was the most important feature. This study provides evidence that restricted maternal nutrition leads to a reduction in the number of cardiomyocytes while melatonin treatment can mitigate some of these disturbances.

Maternal nutrient restriction and over-feeding during gestation alter expression of key factors involved in placental development and vascularization.

Poor maternal nutrition can negatively affect fetal and placental growth and development. However, the mechanism(s) that contribute to altered placenta growth and function are not well understood. We hypothesized that poor maternal diet would impact signaling through the C-X-C motif chemokine ligand (CXCL) 12-CXCL4 axis and/or placental expression of the insulin-like growth factor (IGF) axis. Using our established sheep model of poor maternal nutrition, we examined the effects of restricted- and over-feeding on ewe placentome gene and protein expression. Specifically, ewes were fed a control (CON; 100%), restricted (RES; 60%), or over (OVER; 140%) diet beginning at day 30.2 ± 0.02 of gestation, and samples were collected at days 45, 90, and 135 of gestation, representing periods of active placentation, peak placental growth, and near term, respectively. Placentomes were separated into cotyledon and caruncle, and samples snap frozen. Protein was determined by western blot and mRNA expression by real-time PCR. Data were analyzed by ANOVA and significance determined at P ≤ 0.05. Ewes fed a RES diet had decreased CXCL12 and vascular endothelial growth factor (VEGF), and increased tumor necrosis factor (TNF)α protein compared with CON ewes in caruncle at day 45 (P ≤0.05). In day 45 cotyledon, CXCR7 protein was increased and mTOR was decreased in RES relative to CON (P ≤0.05). At day 90, CXCR4 and CXCR7 were reduced in RES caruncle compared with CON, whereas VEGF was reduced and mTOR increased in cotyledon of RES ewes relative to CON (P ≤0.05). In OVER caruncle, at day 45 CXCR4 and VEGF were reduced and at day 90 CXCR4, CXCR7, and TNFα were reduced in caruncle compared with CON (P ≤0.05). There was no observed effect of OVER diet on protein abundance in the cotyledon (P > 0.05). Expression of IGF-II mRNA was increased in OVER at day 45 and IGFBP-3 was reduced in RES at day 90 in caruncle relative to CON (P ≤0.05). Maternal diet did not alter placentome diameter or weight (P > 0.05). These findings suggest that restricted- and over-feeding negatively impact protein and mRNA expression of key chemokines and growth factors implicated in proper placenta development and function.

259 Effects of Late Gestation Energy and Protein Restriction in Beef Cows on Offspring Growth and Carcass Characteristics

Abstract Our objective was to conduct a systematic literature review and collate data to run a meta-regression on the effect of protein and energy restriction during the last third of gestation on offspring growth and carcass characteristics. The literature search was restricted to only using data from the last third of gestation in spring calving herds such that the last trimester coincided with winter months. Additionally, the search was limited to Bos taurus beef cattle and peer reviewed research articles; review articles and proceeding abstracts/papers were excluded. The search was conducted using the Web of Science. During this process we found contradictory results. As an example, some manuscripts indicate that energy and/or protein restriction during late gestation has a negative impact on offspring growth and/or carcass characteristics; however, there are also manuscripts that report no differences in similar growth and carcass variables (Table 1). Also, most of the studies reported no differences with maternal restriction on offspring body weight at calving and at weaning. One of the limitations in the data set is that most articles did not measure or report dry matter, net energy, or crude protein intake on the dam; therefore, making it difficult to predict the severity of maternal nutrient restriction and the effects on the offspring. Because of this gap in the literature, we suggest that net energy and crude protein intake should be reported in future studies to better determine the effects of maternal restriction during late gestation on offspring growth and carcass characteristics.

Maternal dopamine exposure provides offspring starvation resistance in Daphnia.

The neurotransmitter dopamine has been shown to play an important role in modulating behavioral, morphological, and life history responses to food abundance. However, costs of expressing high dopamine levels remain poorly studied and are essential for understanding the evolution of the dopamine system. Negative maternal effects on offspring size from enhanced maternal dopamine levels have previously been documented in Daphnia. Here, we tested whether this translates into fitness costs in terms of lower starvation resistance in offspring. We exposed Daphnia magna mothers to aqueous dopamine (2.3 or 0 mg/L for the control) at two food levels (ad libitum vs. 30% ad libitum) and recorded a range of maternal life history traits. The longevity of their offspring was then quantified in the absence of food. In both control and dopamine treatments, mothers that experienced restricted food ration had lower somatic growth rates and higher age at maturation. Maternal food restriction also resulted in production of larger offspring that had a superior starvation resistance compared to ad libitum groups. However, although dopamine exposed mothers produced smaller offspring than controls at restricted food ration, these smaller offspring survived longer under starvation. Hence, maternal dopamine exposure provided an improved offspring starvation resistance. We discuss the relative importance of proximate and ultimate causes for why D. magna may not evolve toward higher endogenous dopamine levels despite the fitness benefits this appears to have.