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Hepatic Gene Expression Profiling of Riboflavin-deficient and Riboflavin-rescued Chicken Embryos at 9, 11, 13 and 15 days of incubation

ABSTRACT: This study aimed to identify key genes and pathways related to essential functions of riboflavin during the development of chicken embryos. We used as a unique model a strain of mutant chickens (rd/rd) which is unable to produce a riboflavin-binding protein and lays riboflavin-deficient eggs in which the embryos suddenly die between 13 and 15 days of incubation (e13 and e15, respectively). In these embryos, the diminished activity of flavin-dependent enzymes, particularly those associated with β-oxidation of fatty acid, limits the catabolism of yolk lipids that normally serve as the predominant source of energy required for growth. This malady leads to excessive lipid accumulation in liver and severe hypoglycemia prior to death. This lethal effect can be rescued by an injection of riboflavin just prior to incubation of fertile rd/rd eggs. Microarray-based hepatic transcriptome profiling of riboflavin-deficient (Rf-) and riboflavin-rescue (Rf+) chicken embryos were carried out at 9, 11, 13 and 15 days of incubation. Our microarray analysis identified a dramatic change in gene expression between the Rf- and Rf+ groups on e13 and e15 with 221 and 929 differentially expressed genes (DEGs), respectively. PPAR-gamma, apolipoprotein AV, adipophilin and sterol 14-alpha demethylase were up-regulated, while alpha2 antiplasmin, alpha1 antitrypsin, alpha2 microglobulin, TIP120 and GHRG1 were down-regulated in the Rf- embryos which show an increased lipid accumulation in liver that may be resulted from the disruption in fatty acid beta-oxidation. Genes involved in detoxification, the immune response and blood coagulation were down-regulated, whereas several phosphatase genes were increased. There were no apparent systematic changes in expression of genes associated with the flavin-dependent enzymes, with an exception of an increase in medium chain acyl-CoA dehydrogenase.

ORGANISM(S): Gallus gallus

PROVIDER: GSE96702 | GEO | 2017/11/05

SECONDARY ACCESSION(S): PRJNA379424

REPOSITORIES: GEO

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Project description:Initially, we had compared gene expression differences in the embryonic gonads (E13.5) of the 129 and M19 strains by performing microarray analysis. These studies allowed us to identify downregulation of expression of the D19Bwg1357e, Zfp162 and Cox15 genes in the M19 strain. We next decided to examine gene expression profile changes in the embryonic gonads of 129 and M19 at additional stages so as to identify other TGCT genes from M19 that are also involved in initiation and maintenance of transformed germ cells. For this we isolated E13.5, E15.5 and E17.5 genital ridges from male embryos. Microarray analysis of E13.5, E15.5 and E17.5 genital ridges from male embryos of M19 and 129. Total RNA prepared from the embryonic gonads were analyzed on 12 Affymetrix mouse 430_2 arrays. The objective of the analysis is to determine the differentially expressed genes for the following comparisons: 1. Comparisons of the two strains at each stage, specifically: 1a. 129-E13-GR vs. M19-E13-GR 1b. 129-E15-GR vs. M19-E15-GR 1c. 129-E17-GR vs. M19-E17-GR 2. Comparisons of the different stage in strain 129, specifically: 2a. 129-E13-GR vs. 129-E15-GR 2b. 129-E15-GR vs. 129-E17-GR 3. Comparisons of the different stage in strain CSS, specifically: 3a. M19-E13-GR vs. M19-E15-GR 3b. M19-E15-GR vs. M19-E17-GR After data process and quality assessment, 38069 out of 45101 probe sets proceeded for further analysis to identify differentially expressed genes. Hierarchical cluster analysis shows that samples of two strains from the same stage were grouped together and separated from the other stages (Fig.8). The initial analysis indicates a number of trends in gene expression patterns. Moreover, downregulation of expression of D19Bwg1357e, Zfp162 and Cox15 genes in M19 was also verified in this analysis. These data are currently being analyzed with the help of Dr. Y. Ji and L. Xiao from the Department of Bioinformatics and Computational Biology and Department of Biostatistics at M.D. Anderson Cancer Center. The results are unpublished.

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Hepatic Gene Expression Profiling of Riboflavin-deficient and Riboflavin-rescued Chicken Embryos at 9, 11, 13 and 15 days of incubation

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