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Quantitative evaluation of DNA methylation patterns for ALVE and TVB genes in a neoplastic disease susceptible and resistant chicken model.

Yu Y, Zhang H, Tian F, Bacon L, Zhang Y, Zhang W, Song J - PLoS ONE (2008)

Bottom Line: CpG methylation contents of the ALVE regions were significantly lower in line 6(3) than in line 7(2) in all tissues (P < 0.01) except the ALVE region 3/4 in liver.RNA expressions of ALVE regions 2 and 3 (PPT-U3) were significantly higher in line 6(3) than in line 7(2) (P < 0.01).The methylation level of the promoter region in the TVB was significantly different in blood (P < 0.05) and hypothalamus (P < 0.0001), respectively.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal & Avian Sciences, University of Maryland, College Park, Maryland, USA.

ABSTRACT
Chicken endogenous viruses, ALVE (Avian Leukosis Virus subgroup E), are inherited as LTR (long terminal repeat) retrotransposons, which are negatively correlated with disease resistance, and any changes in DNA methylation may contribute to the susceptibility to neoplastic disease. The relationship between ALVE methylation status and neoplastic disease in the chicken is undefined. White Leghorn inbred lines 7(2) and 6(3) at the ADOL have been respectively selected for resistance and susceptibility to tumors that are induced by avian viruses. In this study, the DNA methylation patterns of 3 approximately 6 CpG sites of four conserved regions in ALVE, including one unique region in ALVE1, the promoter region in the TVB (tumor virus receptor of ALV subgroup B, D and E) locus, were analyzed in the two lines using pyrosequencing methods in four tissues, i.e., liver, spleen, blood and hypothalamus. A significant CpG hypermethylation level was seen in line 7(2) in all four tissues, e.g., 91.86 +/- 1.63% for ALVE region2 in blood, whereas the same region was hemimethylated (46.16 +/- 2.56%) in line 6(3). CpG methylation contents of the ALVE regions were significantly lower in line 6(3) than in line 7(2) in all tissues (P < 0.01) except the ALVE region 3/4 in liver. RNA expressions of ALVE regions 2 and 3 (PPT-U3) were significantly higher in line 6(3) than in line 7(2) (P < 0.01). The methylation levels of six recombinant congenic strains (RCSs) closely resembled to the background line 6(3) in ALVE-region 2, which imply the methylation pattern of ALVE-region 2 may be a biomarker in resistant disease breeding. The methylation level of the promoter region in the TVB was significantly different in blood (P < 0.05) and hypothalamus (P < 0.0001), respectively. Our data disclosed a hypermethylation pattern of ALVE that may be relevant for resistance against ALV induced tumors in chickens.

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mRNA expression difference of TVB in line 63 and line 72.P values indicates the statistical significance for the differences of relative TVB mRNA expression levels between lines 63 and 72 in spleen, liver, and hypothalamus, respectively. A: RT-PCR results. B: Real-time quantitative RT-PCR analysis. n = 5 for each line.
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pone-0001731-g009: mRNA expression difference of TVB in line 63 and line 72.P values indicates the statistical significance for the differences of relative TVB mRNA expression levels between lines 63 and 72 in spleen, liver, and hypothalamus, respectively. A: RT-PCR results. B: Real-time quantitative RT-PCR analysis. n = 5 for each line.

Mentions: The mRNA expression level of the TVB gene in different tissues between the line 63 and line 72 was further examined with real-time quantitative PCR. The results were shown in Figure 9A and 9B. We found that, in general, the mRNA expression levels of the TVB gene were higher in the liver, spleen and hypothalamus in line 63 than in line 72. However, this difference between the two lines was only statistically significant in the hypothalamus (P<0.05), differences were not significant in the spleen (P = 0.15) or the liver (P = 0.14) (Figure 9B).


Quantitative evaluation of DNA methylation patterns for ALVE and TVB genes in a neoplastic disease susceptible and resistant chicken model.

Yu Y, Zhang H, Tian F, Bacon L, Zhang Y, Zhang W, Song J - PLoS ONE (2008)

mRNA expression difference of TVB in line 63 and line 72.P values indicates the statistical significance for the differences of relative TVB mRNA expression levels between lines 63 and 72 in spleen, liver, and hypothalamus, respectively. A: RT-PCR results. B: Real-time quantitative RT-PCR analysis. n = 5 for each line.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2254315&req=5

pone-0001731-g009: mRNA expression difference of TVB in line 63 and line 72.P values indicates the statistical significance for the differences of relative TVB mRNA expression levels between lines 63 and 72 in spleen, liver, and hypothalamus, respectively. A: RT-PCR results. B: Real-time quantitative RT-PCR analysis. n = 5 for each line.
Mentions: The mRNA expression level of the TVB gene in different tissues between the line 63 and line 72 was further examined with real-time quantitative PCR. The results were shown in Figure 9A and 9B. We found that, in general, the mRNA expression levels of the TVB gene were higher in the liver, spleen and hypothalamus in line 63 than in line 72. However, this difference between the two lines was only statistically significant in the hypothalamus (P<0.05), differences were not significant in the spleen (P = 0.15) or the liver (P = 0.14) (Figure 9B).

Bottom Line: CpG methylation contents of the ALVE regions were significantly lower in line 6(3) than in line 7(2) in all tissues (P < 0.01) except the ALVE region 3/4 in liver.RNA expressions of ALVE regions 2 and 3 (PPT-U3) were significantly higher in line 6(3) than in line 7(2) (P < 0.01).The methylation level of the promoter region in the TVB was significantly different in blood (P < 0.05) and hypothalamus (P < 0.0001), respectively.

View Article: PubMed Central - PubMed

Affiliation: Department of Animal & Avian Sciences, University of Maryland, College Park, Maryland, USA.

ABSTRACT
Chicken endogenous viruses, ALVE (Avian Leukosis Virus subgroup E), are inherited as LTR (long terminal repeat) retrotransposons, which are negatively correlated with disease resistance, and any changes in DNA methylation may contribute to the susceptibility to neoplastic disease. The relationship between ALVE methylation status and neoplastic disease in the chicken is undefined. White Leghorn inbred lines 7(2) and 6(3) at the ADOL have been respectively selected for resistance and susceptibility to tumors that are induced by avian viruses. In this study, the DNA methylation patterns of 3 approximately 6 CpG sites of four conserved regions in ALVE, including one unique region in ALVE1, the promoter region in the TVB (tumor virus receptor of ALV subgroup B, D and E) locus, were analyzed in the two lines using pyrosequencing methods in four tissues, i.e., liver, spleen, blood and hypothalamus. A significant CpG hypermethylation level was seen in line 7(2) in all four tissues, e.g., 91.86 +/- 1.63% for ALVE region2 in blood, whereas the same region was hemimethylated (46.16 +/- 2.56%) in line 6(3). CpG methylation contents of the ALVE regions were significantly lower in line 6(3) than in line 7(2) in all tissues (P < 0.01) except the ALVE region 3/4 in liver. RNA expressions of ALVE regions 2 and 3 (PPT-U3) were significantly higher in line 6(3) than in line 7(2) (P < 0.01). The methylation levels of six recombinant congenic strains (RCSs) closely resembled to the background line 6(3) in ALVE-region 2, which imply the methylation pattern of ALVE-region 2 may be a biomarker in resistant disease breeding. The methylation level of the promoter region in the TVB was significantly different in blood (P < 0.05) and hypothalamus (P < 0.0001), respectively. Our data disclosed a hypermethylation pattern of ALVE that may be relevant for resistance against ALV induced tumors in chickens.

Show MeSH
Related in: MedlinePlus