Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

Transcription profiling of mouse liver to investigate the effect of CLA diet on liver gene expression

ABSTRACT: apoE-deficient mice were fed a Western-type diet enriched with linoleic acid (control), cis-9, trans-11-CLA or trans-10, cis-12-CLA (1.0 % wt/wt) for 12 weeks and their hepatic gene expression was analysed using DNA microarrays Experiment Overall Design: Three experimental designs using different genetic models as well as dietary conditions were carried out. In the first experiment, twenty nine three month-old homozygous apoE knockout male mice with a C57BL/6JxOLA129 genetic background, bred at the Unidad Mixta de InvestigaciÃ³n, Zaragoza, were randomly assigned to a control (n=10), c9,t11-CLA-enriched (n=10) or t10,c12-CLA-enriched (n=9) diet. All mice had similar baseline plasma cholesterol concentrations. The mice were fed the different experimental semi-purified diets for 12 weeks. All three diets were isocaloric and isonitrogenous; fat provided 30% of the energy intake. The control diet contained 1.0 % (wt/wt) linoleic acid while the others contained an equivalent amount of either c9,t11-CLA or t10,c12-CLA, as previously described. All diets were prepared by Unilever (Vlaardingen, The Netherlands) and stored in an N2 atmosphere at â??20ÂºC. Fresh food was provided daily

ORGANISM(S): Mus musculus

SUBMITTER: Jesus Osada

PROVIDER: E-GEOD-7076 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Microarray analysis of hepatic gene expression identifies new genes involved in steatotic liver.

Guillén Natalia N Navarro María A MA Arnal Carmen C Noone Enda E Arbonés-Mainar José M JM Acín Sergio S Surra Joaquín C JC Muniesa Pedro P Roche Helen M HM Osada Jesús J

Physiological genomics 20090303 3

<h4>Unlabelled</h4>Trans-10, cis-12-conjugated linoleic acid (CLA)-enriched diets promote fatty liver in mice, while cis-9, trans-11-CLA ameliorates this effect, suggesting regulation of multiple genes. To test this hypothesis, apoE-deficient mice were fed a Western-type diet enriched with linoleic acid isomers, and their hepatic gene expression was analyzed with DNA microarrays. To provide an initial screening of candidate genes, only 12 with remarkably modified expression between both CLA isom ...[more]

PMID: 19258494

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Project description:A summary of the work associated to these microarrays is the following: Diet plays a role in the development of the immune system, and polyunsaturated fatty acids can modulate the expression of a variety of genes. Human milk contains conjugated linoleic acid (CLA), a fatty acid that seems to contribute to immune development. Indeed, recent studies carried out in our group in suckling animals have shown that the immune function is enhanced after feeding 80:20 isomer mix of c9,t11 and t10,c12 CLA, respectively. However, little work has been done on the effects of CLA on gene expression, and even less regarding immune system development in early life. With all these concepts in mind, we determined the expression profile of Mesentheric Lymph Nodes (MLN) from animals supplemented with CLA during gestation and suckling through dam’s milk (Group A) or by oral gavage (Group B), supplemented just during suckling (Group C) and control animals (Group D) with the aid of the specific GeneChip ® Rat Genome 230 2.0 (Affymettrix). Bioinformatic analyses were performed using the GeneSpring GX software package v10.0.2 and lead to the identification of 123 genes differentially expressed in all three dietary approaches. Generation of a Biological Association Network (BAN) evidenced several genes, such as connective tissue growth factor (Ctgf), tissue inhibitor of metalloproteinase 1 (Timp1), galanin (Gal), synaptotagmin 1 (Syt1), growth factor receptor bound protein 2 (Grb2) actin gamma 2 (Actg2) and smooth muscle alpha actin (Acta2), as highly interconnected nodes of the resulting network. Gene underexpression was confirmed by Real-Time RT-PCR. We conclude that Ctgf, Timp1, Gal and Syt1, among others, are genes modulated by CLA supplementation that may have a role on lymphoproliferation and mucosal immune responses in early life. The aim of our study was to evaluate, by using whole genome microarrays, the effects of dietary supplementation with an 80:20 isomer mix of c9,t11 and t10,c12 CLA, respectively, on Mesentheric Lymph Nodes gene expression, during gestation and/or suckling. Three experimental approaches were conducted to assess the effects of CLA supplementation on I) pups from dams fed with 1% CLA diet during the two weeks of gestation and throughout the suckling period (Group A). These pups received CLA through the dam’s milk during suckling (Total Period of CLA Supplementation (TPS) 5 wk); II) pups from dams fed with 1% CLA diet during gestation and standard diet during suckling. These pups were CLA-supplemented daily during suckling by oral gavage (TPS 5 wk) (Group B); III) pups from dams fed standard diet during gestation and suckling and receiving CLA by daily oral gavage throughout the suckling period (TPS 3 wk) (Group C). Group D, pups from dams fed standard diet throughout the study, constituted the reference diet group (TPS 0 wk). Triplicate samples were hybridized for each experimental condition (12 samples in total). The samples provided were analyzed using the specific software GeneSpring GX.

Project description:Cis-9, trans-11-conjugated linoleic acid (c9, t11-CLA), a functional fatty acid, is one of the research hotspots in the fields of functional dairy products development because of its multiple beneficial effects in humans and animals. In milk, most c9, t11-CLA is de novo synthesized from trans-11-octadecenoic acid (TVA) and it is confirmed that genes such as stearoyl-coA desaturase 1 (SCD1) play a key role in the synthesis. However, few studies have been reported to deeply elucidate the SCD1-dependent molecular mechanism of cis9, trans11-CLA synthesis and its relationship with the pathways of energy metabolism and lipid metabolism. Therefore, in the present study, MAC-T cells were divided into three groups: CAY group (SCD1-inhibited MAC-T cell model with the addition of CAY, TVA), TVA group (only TVA), and Control group (without CAY, TVA). The relative mRNA expression of SCD1 was measured using real-time PCR, while TVA accumulation and c9, t11 -CLA synthesis were analyzed using gas chromatography (GC). The SCD1-related proteins were firstly screened in MAC-T cells by Tandem Mass Tag (TMT)-based quantitative proteomics analysis, then their functions were annotated by bioinformatic analysis, and finally their relationships with SCD1 were evaluated by parallel reaction monitoring (PRM) analysis and small RNA interference. The results showed that the deficiency of SCD1 led by CAY10566 blocked the synthesis of c9, t11-CLA in MAC-T cells. Sixty-one SCD1-associated proteins were screened and found to be mainly involved in the pathways of energy metabolism and lipid metabolism, such as glycolytic pathway, pentose phosphate pathway, fatty acid elongation pathway, and unsaturated fatty acid biosynthesis. Among these genes, 17 proteins were validated under the PRM analysis. PGAM1 (phosphoglycerate mutase 1), TPI1 (triosephosphate isomerase 1), LDHB (lactate dehydrogenase B), and ALDOA (aldolase, fructose-bisphosphate A) were verified to have a negative relationships with SCD1. This study furthered our understanding of the molecular mechanisms of c9, t11-CLA synthesis in the mammary glands of dairy cows.

Project description:Conjugated linoleic acid (CLA), a class of fatty acids found in beef and dairy products, has been shown to inhibit tumorigenesis in a variety of cancer model systems. Based on previously well-documented anti-tumor activity of CLA in rodent models of breast cancer, a pilot study was initiated to examine the effect of dietary CLA in a well-established transgenic model of breast cancer. Western blots were performed for the detection of AKT, c-Src, ERK1/2, and Cdc24. CLA significantly increased tumor burden (p<0.1) independent of an increase in oncogenic signaling. Mammary gland whole mounts indicated a loss of mammary adipose and extensive epithelial expansion in CLA-treated animals. Microarray analysis indicated a significant reduction in cytoskeletal related genes with at least a two-fold decrease in five out of six CLA-fed animals compared to untreated controls. Reduction of Cdc42, a key regulator of cell adhesion and cytoskeletal arrangements, was confirmed at the protein level by western blot (p<0.01). These findings suggest that dietary CLA may advance the malignant phenotype by promoting a loss of cell polarity and adhesion in the mammary gland epithelium. This action may have serious clinical implications for a subset high-risk population and warrants further investigation. Virgin, four-week-old PyV-mT mice were administered a diet of a mixed-isomer CLA formulation (1% wt/wt) (N=6) or control AIN96G diet (N=5) for four weeks. Measurements of food disappearance, weights and palpations were recorded weekly. All animals were euthanized at eight weeks of age. Formalin-fixed, paraffin-embedded mammary gland tissue was used for H&E and trichrome staining and immunohistochemistry for Ki67. Tissue levels of CLA were measured by gas chromatography. Thoracic mammary glands were fixed in glacial acetic acid:ethanol and carmine stained. cDNA microarray was performed on RNA from 6 CLA-fed mice and 4 control mice using the Affymetrix 430 2.0 mouse genome chips.

Dataset Information

Transcription profiling of mouse liver to investigate the effect of CLA diet on liver gene expression

Publications

Microarray analysis of hepatic gene expression identifies new genes involved in steatotic liver.

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