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| The Effects of Force-Feeding on DNA Methylation and mRNA Expression of SCD5, SREBP2 and ELOVL6 in Landes(Anser anser) |
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Abstract DNA methylation is a hotspot in epigenetic research. This study was aimed to explore whether the force-feeding could change the levels of DNA methylation and gene expression related to lipid metabolism in liver of Landes(Anser anser), and to investigate the relationship between DNA methylation and gene expression. The levels of DNA methylation of stearovl-CoA desaturase 5 gene (SCD5), sterol regulatory element-binding protein 2 gene (SREBP2) and elongase of very long chain fatty acids 6 gene (ELOVL6) were analyzed with pyrosequencing method and quantified by qRT-PCR. The upstream promoter sequence of SCD5 (about 2 800 bp) was downloaded from University of California Santa Cruz (UCSC) website, there were 2 CpG islands, and the percentage of cytosine, G+C and CpG island were respectively reached of 22.7%, 50.06% and 3.81% after analyzing from the 406~1 220 bp CpG island (full-length fragment of 815 bp). The upstream promoter sequence of SREBP2 (about 2 248 bp) was also downloaded from UCSC website, there were 2 CpG islands, and the percentage of cytosine, G+C and CpG island were respectively reached of 24.87%, 56.61% and 5.29% after analyzing from the 1~965 bp CpG island (full-length fragment of 965 bp). The upstream promoter sequence of ELOVL6 (about 2 248 bp) was downloaded from UCSC website, there was 1 CpG island, and the percentage of cytosine, G+C and CpG island were respectively reached of 37.14%, 55.71% and 4.30% after analyzing from the 1 426~1 775 bp CpG island (full-length fragment of 350 bp). A total of 23 CpGs related DNA methylation were detected by using pyrosequencing method after amplified the promoter sequence of CpG island of SCD5, SREBP2 and ELOVL6 (each gene had two fragments of 37~55 bp). Results showed that the levels of DNA methylation of the 2 fragments of SCD5, SREBP2 and ELOVL6 in force-feeding group (FFG) and control group(CG) were respectivlely higher than 20%, belongs to high methylation status. The levels of DNA methylation of CG were greater than FFG. Significant difference were found on DNA methylation in the first fragment of SCD5 and the first fragment of SREBP2 between FFG and CG(P＜0.05), markedly significant difference (P＜0.01) in the second fragment of SREBP2, but no significant difference(P＞0.05) in the second fragment of SCD5 and the first fragment and the second fragment of ELOVL6 between FFG and CG. The mRNA expression of mRNA of SCD5 and ELOVL6 in FFG were greater than those of CG, and there relative gene expression of SREBP2 in FFG was lower than that of CG (P＜0.05). In conclusion, force-feeding decreased the expression of DNA methylation of SCD5, SREBP2 and ELOVL6, increased the mRNA expression of SCD5 and ELOVL6 in liver of goose, but decreased the mRNA expression of SREBP2, which indicated that the methylation of SCD5 and ELOVL6 played a negative regulation effect on the mRNA expression of SCD5 and ELOVL6. The result will provide a reference to the research method of DNA methylation on epigenetics and an important molecular genetics basis on breeding high quality and high yield goose fatty liver, it will also provide theoretical basis for researching the metabolic diseases such as obesity and fatty liver.
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Received: 23 March 2016
Published: 06 August 2016
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