Identifying gene expression differences induced by diets that lead to higher omega-3 fatty acid deposition in beef cows
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ABSTRACT: The objective of this project is identifying differentially expressed (DE) genes which are associated with higher omega-3 fatty acids deposition in beef cows. Omega-3 fatty acids have been found to influence meat flavor and are beneficial to human health. Supplementation of livestock diets with flaxseed, a rich source of ë±-linolenic acid, is the most common means of producing omega-3 fatty acid-enriched animal products. Towards the goal of enhancing beef fatty acid composition, 64 crossbred cull cows (~30 months of age) with similar breed composition were randomized by weight/body condition, and fed one of four 50:50 forage:concentrate diets on a DM basis (16 cows/treatment), containing ground barley grain with either hay or silage, supplemented with 0 or 15% ground flaxseed (DM basis). Cows were slaughtered after spending 140 days on the treatment diets. Five cows from each of the four diets were selected for transcriptional analysis based on FA profiles of the kidney fat collected at slaughter. RNA was isolated from Longissimus thoracis muscle, subcutaneous and kidney fat of each cow (20 samples/tissue) and hybridized in duplicate to BOMC 24K 60-mer microarrays. Differential gene expression between flax-fed and non-flax-fed cows as well as identifying those genes associated with fatty acid metabolism were studied.
Project description:Intramuscular fat (IMF) storage is a biological process with strong impact on nutritional and technological properties of meat, and also with relevant consequences on human health. The genetic architecture of IMF content and composition phenotypes has been thoroughly studied in pigs through the identification of quantitative trait loci (QTL) and the estimation of genetic parameters. A question that has not been elucidated yet is if the genetic determinants of IMF-related phenotypes are muscle specific or, conversely, they have broad effects on the whole skeletal muscle compartment. We have addressed this question by generating lipid QTL maps for two muscles with a high commercial value, gluteus medius (GM) and longissimus thoracis et lumborum (LTL), in a Duroc commercial population (N=350). As a complementary approach, we have analysed the mRNA expression pattern of both muscles at a whole genome scale. The lack of concordance between the GM and LTL QTL maps evidenced that the effects of polymorphisms influencing IMF, cholesterol and fatty acids contents are modulated to some extent by complex spatial factors related with muscle location, metabolism and function. This interpretation was supported by our finding that genes influencing cell differentiation, muscle development and function and lipid metabolism are differentially expressed between muscles. These results have important implications on the implementation of genomic selection schemes aimed to improve the lipid profile of swine meat. Moreover, they confirm pigs as a valuable model to dissect the genetic basis of muscle lipid phenotypes of clinical interest in human. Longissimus thoracis et lumborum muscle tissue from 10 high and 9 low fattening Duroc pigs were compared in this study.
Project description:In the last 20 years, there has been significant research towards defining the genetic basis of lipid metabolism and meat quality related traits in pigs. Nowadays, the study of the transcriptome and its regulatory mechanisms allows going far beyond in the genetic dissection of these complex traits. In present study, a genome-wide eQTL scan aiming to detect pig genome regions regulating levels of skeletal muscle mRNA expression has been performed. This study has been conducted on a commercial Duroc population where a number of QTL for muscle fat deposition and fatty acid composition had been detected. GeneChip Porcine Genome® arrays (Affymetrix) were used to determine the gene expression levels of gluteus medius samples from 105 Duroc pigs belonging to two groups with divergent phenotypes for fatness traits. This experimental design aimed to favour detection of eQTL affecting genes related to lipid metabolism and meat quality traits. The whole genome scan with a panel of 110 microsatellites allowed us detecting 613 genome-wide significant eQTL unequally distributed across the pig genome, SSC5 and SSC3 harbouring the highest number of eQTL. Moreover, 11 genome regions with eQTL affecting the expression levels of a high number of genes (eQTL hot spots) have been described. After mapping target probes and discarding low quality probes, a total of 59 cis- and 396 trans-acting eQTL were retained for further analyses. The functional classification showed that lipid-related GO terms were not the most enriched by the list of eQTL-regulated genes. However, a number of regulated genes functionally related to lipid metabolism and fat deposition traits were identified, and their functional relationship with these phenotypes were further investigated. With this purpose, eQTL results were integrated with 1) QTL linkage maps and 2) correlation data between phenotypes and gene expression levels. As a result, a comprehensive list of 29 positional and functional candidate genes was elaborated. These results represent a valuable contribution to the comprehension of genetic regulation of skeletal muscle individual gene expression in swine species, and a first step towards disentangling gene networks and molecular mechanisms involved in lipid metabolism and meat quality traits. 105 gluteus medius samples from 105 animals belonging to two groups of 53 and 52 animals each: HIGH group had higher carcass, plasma and muscle fat content; LOW group had lower carcass, plasma and muscle fat content
Project description:The objective of this study was to identify the molecular mechanisms and biological pathways associated with the anticancer effects of flaxseed (richest plant source of Omega-3 fatty acid) in laying hen model of ovarian cancer. Study shows a significant reduction in the severity of the disease and increased survival of the laying hens fed with flaxseed. 2 X 2 condition experiment, Diet (Control & Flaxseed fed) and Tissue (Normal & Cancer). Biological replicates: 6 control normal replicates, 6 control cancer replicates, 6 flaxseed normal replicates and 6 flaxseed cancer replicates.
Project description:The optimal ratio of omega-6 to omega-3 polyunsaturated fatty acids (PUFAs) is important for keeping homeostasis of biological processes and metabolism, yet the underlying biological mechanism is poorly understood. The objective of this study was to identify changes in the pig liver transcriptome induced by a diet enriched with omega-6 and omega-3 fatty acids, and to characterize the biological mechanisms related to PUFA metabolism. Polish Landrace pigs (n =12) were fed diet enriched with linoleic acid (LA, omega-6) and alpha-linolenic acid (ALA, omega-3 family) or standard diet as a control. The fatty acids profiling was assayed in order to verify how feeding influenced the fatty acids content in liver, and subsequently next-generation sequencing (NGS) was used to identify differentially expressed genes (DEG) between transcriptomes between dietary groups. The biological mechanisms and pathway interaction networks were identified by analysis in DAVID and Cytoscape tools. Fatty acids profile analysis indicated a higher contribution of PUFAs in liver for LA and ALA-enriched diet group, particularly for the omega-3 fatty acids family, but not omega-6. Next-generation sequencing identified 3,565 DEG, 1,484 of which were induced and 2,081 were suppressed by PUFA supplemenation. Low ratio of omega-6/-3 fatty acids resulted in modulation of fatty acids metabolism pathways and over-representation of genes involved in membrane composition, signal transduction and immune response pathways. In conclusion, a diet enriched with omega-6 and omega-3 fatty acids altered the transcriptomic profile of the pig liver and affected a set of genes involved in metabolic pathways important to animal health status. Hepatic mRNA profiles of Polish Landrace pig breed fed two different diets, were generated by deep sequencing, using Illumina MiSeq. Experimental diet was enriched with polyunsaturated fatty acids (omega-6 and omega-3), while standard diet remain as a cotrol. 2 pooled samples each containing RNA extracts from 6 individuals livers were analyzed.
Project description:Omega-3 and omega-6 polyunsaturated fatty acids (PUFA) have important signalling roles in the body. The goal of this study was to investigate the impact of linoleic acid (LA, omega-6) and alpha-linolenic (ALA, omega-3) on global skeletal muscle gene expression. We were also interested to study the impact of these fatty acids on myokine expression. To differentiate the roles of essential dietary PUFA on skeletal muscle function, we fed male rats a control diet (AIN-93G) or diets containing 10% safflower oil or flaxseed oil. Skeletal muscle gene expression was investigated by microrray.
Project description:The current study was designed to determine if dietary fatty acid concentration and composition affects the development and progression of nonalcoholic fatty liver disease. Male SD rats were overfed diets low (5%) or high (70%) fat diets via total enteral nutrition where the fat source was olive oil (monounsaturated), or corn oil (polyunsaturated). Overfeeding 5% corn oil produced little steatosis relative to feeding 5% olive oil. This was associated with lower fatty acid synthesis and reduced SREBP-c signaling in the 5% corn oil group. Overfeeding 70% fat diets increased steatosis and lead to increased liver necrosis in the 70% corn oil but not olive oil group. Increased injury after feeding polyunsaturated fat diets was linked to peroxidizability of hepatic free fatty acids and triglycerides and appearance of peroxidaized lipid products HETES and HODES previously linked to clinical nonalcoholic steatohepatitis. Male SD rats were overfed diets low (5%) or high (70%) fat diets via total enteral nutrition where the fat source was olive oil (monounsaturated) or corn oil (polyunsaturated).
Project description:Dietary supplementation with ω-3 polyunsaturated fatty acids (ω-3 PUFAs), specifically the fatty acids docosahexaenoic acid (DHA; 22:6 ω-3) and eicosapentaenoic acid (EPA; 20:5 ω-3), is known to have beneficial health effects including improvements in glucose and lipid homeostasis and modulation of inflammation. To evaluate the efficacy of two different sources of ω-3 PUFAs, we performed gene expression profiling in the liver of mice fed diets supplemented with either fish oil or krill oil. We found that ω-3 PUFA supplements derived from a phospholipid krill fraction (krill oil) downregulated the activity of pathways involved in hepatic glucose production as well as lipid and cholesterol synthesis. The data also suggested that krill oil-supplementation increases the activity of the mitochondrial respiratory chain. Surprisingly, an equimolar dose of EPA and DHA derived from fish oil modulated fewer pathways than a krill oil-supplemented diet and did not modulate key metabolic pathways regulated by krill oil, including glucose metabolism, lipid metabolism and the mitochondrial respiratory chain. Moreover, fish oil upregulated the cholesterol synthesis pathway, which was the opposite effect of krill supplementation. Neither diet elicited changes in plasma levels of lipids, glucose or insulin, probably because the mice used in this study were young and were fed a low fat diet. Further studies of krill oil supplementation using animal models of metabolic disorders and/or diets with a higher level of fat may be required to observe these effects. Twenty-one microarrays: three diets (CO, FO, KO) x seven mice per diet x one microarray per mouse
Project description:Early embryonic development is enhanced in Holstein cows fed diets enriched in specific polyunsaturated fatty acids. However, the molecular mechanisms affected by specific polyunsaturated fatty acids during early embryonic development in cattle are poorly understood. Therefore, our objective was to evaluate the maternal effects of diets enriched in linoleic or α-linolenic acid on transcriptome profiling of in vivo bovine pre-implantation embryos.
Project description:Flaxseed (FS), a nutritional supplement consisting mainly of omega-3 fatty acids and lignan phenolics, has potent anti-inflammatory, anti-fibrotic and antioxidant properties. We have shown that dietary FS supplementation ameliorates oxidative stress and inflammation in experimental models of acute and chronic lung injury in mice resulting from diverse toxicants. The development of lung tissue damage in response to direct or indirect oxidant stress is a complex process, associated with changes in expression levels of a number of genes. We therefore postulated that flaxseed may modulate gene expression of vital signaling pathways, thus interfering with the development of tissue injury. We evaluated gene expression in lungs of flaxseed-fed (10% FS) mice under unchallenged, control conditions and 48hrs post-radiation treatment (13.5 Gy). Gene expression levels in lung tissues were analyzed using three arrays for each sample, whereby a total of 28,800 genes were evaluated. C57BL/6 black mice were fed a 10% flaxseed diet for 21 days and/or given 13.5Gy thoracic radiation and sacrificed 48hrs post radiation.
Project description:Flaxseed (FS), a nutritional supplement consisting mainly of omega-3 fatty acids and lignan phenolics, has potent anti-inflammatory, anti-fibrotic and antioxidant properties. We have shown that dietary FS supplementation ameliorates oxidative stress and inflammation in experimental models of acute and chronic lung injury in mice resulting from diverse toxicants. The development of lung tissue damage in response to direct or indirect oxidant stress is a complex process, associated with changes in expression levels of a number of genes. We therefore postulated that flaxseed may modulate gene expression of vital signaling pathways, thus interfering with the development of tissue injury. We evaluated gene expression in lungs of flaxseed-fed (10% FS) mice under unchallenged, control conditions and 48hrs post-radiation treatment (13.5 Gy). Gene expression levels in lung tissues were analyzed using three arrays for each sample, whereby a total of 28,800 genes were evaluated. C57BL/6 black mice were fed a 10% flaxseed diet for 21 days and/or given 13.5Gy thoracic radiation and sacrificed 48hrs post radiation.