Transcriptomic analysis of sheep hypothalamus discloses regulatory genes potentially involved in sex-dependent differences in body weight of progeny born to dams supplemented with omega-3 fatty acids or methionine during late-gestation.

Fetal programming research conducted in sheep has reported sexually dimorphic responses on growth of the progeny born to in-utero methionine or omega-3 fatty acids supplementation. However, the biological mechanism behind the nutrient by sex interaction as a source of variation in offspring body weight is still unknown. A high-throughput RNA sequencing data of hypothalamus samples from 17 lambs were used in the current study to identify differentially expressed genes (DEGs) between males and females born to dams supplemented with different nutrients during late-gestation. Ewes received a basal diet without omega-3 fatty acids or methionine supplementation as the control (CONT); omega-3 fatty acids supplementation (FAS), or methionine supplementation (METS). A list of regulated genes was generated. Data was compared as CONT vs. FAS and CONT vs. METS. For CONT vs. METS, a treatment by sex interaction was found (adjusted P-value < 0.05) on 121 DEGs (112 upregulated and 9 downregulated) on female lambs born to METS compared with METS males. Importantly, with the sex interaction term, more than 100 genes were upregulated in female lamb's hypothalamuses born to METS. Gene Ontology (GO) and Ingenuity Pathway Analysis (IPA) were performed using the DEGs from female lambs. Terms under biological process (related to morphogenesis, organism, and tissue development), cellular component (related to chromatin, extracellular components), and molecular function (involved in chromatin structure and transcription and factors linked to binding DNA) were presented (adjusted P-value < 0.05) for GO. For the IPA, the top-scoring network was developmental disorder, endocrine system development and function, and organ morphology. Only a few differences were observed in the comparison between the interaction of sex and treatment for the CONT vs. FAS comparison. The markedly increased number of DEGs substantially involved in developmental and growth processes indicates the extent to which maternal methionine supplementation causes the sexually dimorphic effects observed in the offspring.

Effects of maternal dietary fatty acids during mid-gestation on growth, glucose metabolism, carcass characteristics, and meat quality of lamb progeny that were fed differing levels of dry matter of intake.

This experiment evaluated growth, glucose metabolism, carcass characteristics, and meat quality of market lambs that were offered ad libitum or restricted (85% of ad libitum) feed intake following two different maternal fatty acid (FA) supplementations while in-utero. Ewes received either a diet supplemented with polyunsaturated FA or saturated/monounsaturated FA during mid- to late-gestation. Following weaning, progeny wethers were fed either ad libitum or a restricted level of feed intake. Ewe FA supplementation did not affect (P ≥ 0.11) growth, meat quality, nor plasma glucose or insulin concentrations of the progeny. Carcass body fat and yield grade of the progeny were affected (P = 0.01) by maternal FA supplementation and restricted feed intake. In summary, maternal FA supplementation did not affect progeny growth, while feed restriction during finishing did not affect meat quality. The interaction between maternal FA supplementation and finishing strategy for body fat accretion indicates that metabolism and the supply of FA during gestation may warrant further investigation.

Maternal supply of a source of omega-3 fatty acids and methionine during late gestation on the offspring's growth, metabolism, carcass characteristic, and liver's mRNA expression in sheep.

The objective of the present experiment was to evaluate the effect of maternal supplementation with fatty acids (FAs) and methionine (Met) during late gestation on offspring growth, energy metabolism, plasma resolvin (RvD1) concentration, carcass characteristics, and hepatic mRNA expression. Ewes (5 pens/treatment; 3 ewes/pen) blocked by body weight (BW) were assigned to one of four treatments from day 100 of gestation until lambing. The treatments were: basal diet (NS) without FAs or Met supplementation; FA supplementation (FS; 1.01 % of Ca salts, containing n-3 FA); Met supplementation (MS; 0.1 % of rumen-protected methionine); and FS and MS (FS-MS). At birth (day 0), ewes and lambs were placed in a common pen. On day 60, lambs were weaned, sorted by sex, blocked by BW, and placed on a common finishing diet for 54 d (FP). A lamb per pen was used for a glucose tolerance test (GTT) after the FP. Carcass characteristics were recorded on day 56. Lamb data were analyzed as a randomized complete block design with a 2 × 2 × 2 factorial arrangement, with repeated measurements when needed (SAS 9.4). At weaning, lambs born to MS- or FS-fed ewes were heavier than lambs born from FS-MS ewes (FS × MS × Time; P = 0.02). A marginal significant FS × MS interaction (P = 0.09) was also observed on RvD1; lambs born to ewes in the NS and FS-MS treatments showed a lower RvD1 plasma concentration when compared with lambs born to FS- or MS-fed ewes. Lambs born to dams fed FA showed an increase (P = 0.05) in liver COX-2 mRNA relative expression. Lambs born to ewes supplemented with Met showed an increase (P = 0.03) in liver FABP4 mRNA expression. An FS × MS × Time interaction (P = 0.07) was observed in plasma glucose during the GTT; lambs born from FS-fed ewes showed lower plasma glucose concentration than lambs born to Met-supplemented ewes at 2 min after bolus administration. During the GTT, a marginal significant effect (P = 0.06) was observed for the lamb average insulin concentration due to maternal Met supplementation during late gestation, where these lambs had the lowest plasma concentration. Contrary to our hypothesis, the interaction of FA and Met supplementation during late gestation did not show a greater positive effect on offspring postnatal growth and metabolism. However, the individual supplementation of each nutrient has an effect on offspring development with a concomitant change in markers involved in the inflammatory response and energy metabolism.

Late gestation supplementation of omega-3 fatty acids (FAs) or methionine (Met) alters the offspring's development. However, the effect of both nutrients on the physiology and growth of the progeny has not been explored. The experiment's objective was to evaluate the effect of dam supplementation with a Met and the omega-3 FAs, such as eicosapentaenoic and docosahexaenoic acids, during late gestation on growth, energy metabolism, and inflammatory response markers of the lamb. Ewes received one of the four following treatments: basal diet without FA or Met, FA supplementation, Met supplementation, or FA and Met supplementation. Supplementation of omega-3 FAs and Met did not show a greater effect on postnatal growth and metabolism of the offspring compared with the supplementation of each nutrient individually. However, individual supplementation influences offspring development with a concomitant change in markers involved in the inflammatory response and energy metabolism. Results of the present experiment suggest that offspring born to mothers that were fed FAs or Met on late gestation could have the ability to better cope with inflammatory processes, which could improve their long-term growth performance. Moreover, maternal supplementation of Met during late gestation could modulate offspring's glucose­insulin system, which may also affect offspring's growth.

Role of Long Chain Fatty Acids in Developmental Programming in Ruminants.

Nutrition plays a critical role in developmental programs. These effects can be during gametogenesis, gestation, or early life. Omega-3 polyunsaturated fatty acids (PUFA) are essential for normal physiological functioning and for the health of humans and all domestic species. Recent studies have demonstrated the importance of n-3 PUFA in ruminant diets during gestation and its effects on pre-and postnatal offspring growth and health indices. In addition, different types of fatty acids have different metabolic functions, which affects the developmental program differently depending on when they are supplemented. This review provides a broad perspective of the effect of fatty acid supplementation on the developmental program in ruminants, highlighting the areas of a developmental program that are better known and the areas that more research may be needed.

Maternal Supply of Fatty Acids during Late Gestation on Offspring's Growth, Metabolism, and Carcass Characteristics in Sheep.

Lambs born from dams supplemented with different sources of fatty acids (FA) during late gestation have a different growth rate and plasma glucose concentration. The main objectives of this experiment were to evaluate the effect of supplementing different sources of FA during late gestation on offspring plasma metabolite concentrations, growth, and on a glucose tolerance test (GTT) during the finishing phase. Fifty-four lambs (18 pens, 3 lambs/pen) were born from ewes supplemented during late gestation with one of three treatments: (1) no FA (NF); (2) a source of monounsaturated FA (PDS, 1.01% of Ca salts); or (3) a source of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (EDS, 1.01% of Ca salts containing). At birth (day 0), supplementation ceased, and all ewes and lambs were placed in a common pen. On day 60, lambs were weaned, grouped by sex, blocked by body weight (BW), and placed on a common finishing diet for 54 days (FP). One lamb per pen was used for the GTT after the FP. There was a tendency for FA × Sex × Day interaction (p = 0.08) on lamb growth during the finishing period, with PDS females being heavier than PDS males, while EDS males were heavier than EDS females at day 60. There was a tendency for FA × Sex interaction (p = 0.06) for plasma insulin concentration for the GTT. Plasma insulin concentration of wethers increased as FA unsaturation degree increased during the GTT; the opposite happened with the plasma insulin concentration of female lambs. In conclusion, FA supplementation during late gestation tended to modified growth and insulin response to a GTT; these changes differed with the degree of FA unsaturation of the supplement and lamb sex.

Effects of maternal dietary omega-3 polyunsaturated fatty acids and methionine during late gestation on fetal growth, DNA methylation, and mRNA relative expression of genes associated with the inflammatory response, lipid metabolism and DNA methylation in placenta and offspring's liver in sheep.

BACKGROUND: Omega-3 PUFA or methionine (Met) supply during gestation alters offspring physiology. However, the effect of both nutrients on fetal development has not been explored. Our objective was to determine the effects of supplementation of these two nutrients during late gestation on fetal growth, DNA methylation, and mRNA expression of genes associated with the inflammatory response, and DNA methylation. Ewes (n = 5/treatment) were fed from day 100 to 145 of gestation one of the following treatments: 1) basal diet (NS) without fatty acids (FS) or methionine (MS) supplementation; 2) FS (10 g/kg Ca salts, source omega-3 PUFA); 3) MS (1 g/kg rumen protected methionine); and 4) FS and MS (FS-MS). On day 145, ewes were euthanized, and data from dams and fetus was recorded. Placenta (cotyledon), fetal liver, and blood samples were collected. RESULTS: A treatments interaction on fetal liver weight, ewe body weight and body condition score (BCS) was observed; FS-MS were heavier (P <  0.01) than FS and MS, and FS-MS ewes had a better (P = 0.02) BCS than NS. Methionine increased (P = 0.03) ewe plasma glucose concentration. Fetal liver global DNA methylation increased (P <  0.01) in FS and MS. Dietary treatments modify the mRNA relative expression on some of the genes evaluated. In the fetal liver, FS increased (P = 0.04) the mRNA relative expression of arachidonate-5-lipoxygenase-activating-protein and tended to decrease (P = 0.06) methionine-adenosyltransferase-1A. Moreover, MS decreased (P = 0.04) DNA-methyltransferase-1 and tended to decrease (P = 0.08) free-fatty-acid-receptor-1 mRNA relative expression. Furthermore, FS-MS decreased mRNA relative expression of tumor-necrosis-factor-alpha (P = 0.05), peroxisome-proliferator-activated-receptor-delta (P = 0.03) and gamma (P = 0.04), tended to decrease (P ≤ 0.09) interleukin-6, fatty-acid-transport-protein-1, and delta-5-desaturase, and increased adenosylhomocysteinase (P = 0.04) mRNA relative expression. In cotyledon, FS tended to decrease fatty acid binding protein 4 (P = 0.09) mRNA relative expression. CONCLUSION: Omega-3 PUFA and Met supplementation improves dam's performance in late gestation, which was positively correlated with an increase in offspring's liver development. Moreover, FS-MS decreased mRNA relative expression of proinflammatory cytokines, and lipogenic genes, and increased the expression on an enzyme that has an important role in methylation.