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Association of DNA Methylation Levels with Tissue-specific Expression of Adipogenic and Lipogenic Genes in Longissimus dorsi Muscle of Korean Cattle.

Baik M, Vu TT, Piao MY, Kang HJ - Asian-australas. J. Anim. Sci. (2014)

Bottom Line: DNA methylation levels of all five CpG sites from the FABP4 gene promoter region were also lower (p<0.001) in the IMF than in the muscle portion.Thus, mRNA levels of both PPARG1 and FABP4 genes were inversely correlated with DNA methylation levels in regulatory regions of CpG sites of the corresponding gene.Our findings suggest that DNA methylation status regulates tissue-specific expression of adipogenic and lipogenic genes in the IMF and muscle portions of LM tissue in Korean cattle.

View Article: PubMed Central - PubMed

Affiliation: Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, College of Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Korea ; Institute of Green Bio Science Technology, Pyeungchang 232-916, Korea .

ABSTRACT
Epigenetic factors, such as DNA methylation status, may regulate adipogenesis and lipogenesis, thus affecting intramuscular fat (IMF) deposition in longissimus dorsi muscle (LM) of beef cattle. In Korean cattle steers, the LM consists mainly of muscle tissue. However, the LM tissue also contains IMF. We compared the gene expression levels between the IMF and muscle portions of the LM after tissue separation. Real-time polymerase chain reaction analysis showed that the mRNA levels of both adipogenic peroxisome proliferator-activated receptor gamma isoform 1 (PPARG1) and lipogenic fatty acid binding protein 4 (FABP4) were higher (p<0.01) in the IMF than in the muscle portion of the LM. We determined DNA methylation levels of regulatory regions of the PPARG1 and FABP4 genes by pyrosequencing of genomic DNA. DNA methylation levels of two of three CpG sites in the PPARG1 gene promoter region were lower (p<0.05) in the IMF than in the muscle portion of the LM. DNA methylation levels of all five CpG sites from the FABP4 gene promoter region were also lower (p<0.001) in the IMF than in the muscle portion. Thus, mRNA levels of both PPARG1 and FABP4 genes were inversely correlated with DNA methylation levels in regulatory regions of CpG sites of the corresponding gene. Our findings suggest that DNA methylation status regulates tissue-specific expression of adipogenic and lipogenic genes in the IMF and muscle portions of LM tissue in Korean cattle.

No MeSH data available.


Related in: MedlinePlus

Target area (a), DNA methylation levels (b), and mRNA levels (c) of the peroxisome proliferator activated receptor gamma 1 (PPARG1) gene in intramuscular fat (IMF) and muscle portion of Korean cattle steer longissimus dorsi muscle tissue. (a) DNA methylation assay target area of the PPARG1 gene promoter region and transcription factor binding sites. TFCP2, transcription factor CP2. (b) DNA methylation levels were determined by pyrosequencing of bisulfite-treated DNA. Values are the mean+SE (n = 5). (c) The mRNA levels were determined by real-time PCR and normalized against a housekeeping gene. Muscle portion data were normalized to 1.0. Values are the mean+SE (n = 10). * p<0.05; ** p<0.01. PCR, polymerase chain reaction; SE, standard errors.
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f1-ajas-27-10-1493: Target area (a), DNA methylation levels (b), and mRNA levels (c) of the peroxisome proliferator activated receptor gamma 1 (PPARG1) gene in intramuscular fat (IMF) and muscle portion of Korean cattle steer longissimus dorsi muscle tissue. (a) DNA methylation assay target area of the PPARG1 gene promoter region and transcription factor binding sites. TFCP2, transcription factor CP2. (b) DNA methylation levels were determined by pyrosequencing of bisulfite-treated DNA. Values are the mean+SE (n = 5). (c) The mRNA levels were determined by real-time PCR and normalized against a housekeeping gene. Muscle portion data were normalized to 1.0. Values are the mean+SE (n = 10). * p<0.05; ** p<0.01. PCR, polymerase chain reaction; SE, standard errors.

Mentions: To determine DNA methylation levels, target regions of PPARG1 (Figure 1a) and FABP4 (Figure 2a) genes were selected from the CpG islands, which were searched using CpG Island Searcher (USC Norris Comprehensive Cancer Center, USA; http://www.uscnorris.com/cpgislands2/cpg.aspx). Transcription factor binding sites (Figures 1a and 2a) were determined using TFSEARCH (Computational Biology Research Center, National Institute of Advanced Industrial Science and Technology, Japan; http://www.cbrc.jp/research/db/TFSEARCH.html). We tried to find promoter regions of the genes in which important transcription factor binding sites were located that may regulate gene expression.


Association of DNA Methylation Levels with Tissue-specific Expression of Adipogenic and Lipogenic Genes in Longissimus dorsi Muscle of Korean Cattle.

Baik M, Vu TT, Piao MY, Kang HJ - Asian-australas. J. Anim. Sci. (2014)

Target area (a), DNA methylation levels (b), and mRNA levels (c) of the peroxisome proliferator activated receptor gamma 1 (PPARG1) gene in intramuscular fat (IMF) and muscle portion of Korean cattle steer longissimus dorsi muscle tissue. (a) DNA methylation assay target area of the PPARG1 gene promoter region and transcription factor binding sites. TFCP2, transcription factor CP2. (b) DNA methylation levels were determined by pyrosequencing of bisulfite-treated DNA. Values are the mean+SE (n = 5). (c) The mRNA levels were determined by real-time PCR and normalized against a housekeeping gene. Muscle portion data were normalized to 1.0. Values are the mean+SE (n = 10). * p<0.05; ** p<0.01. PCR, polymerase chain reaction; SE, standard errors.
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f1-ajas-27-10-1493: Target area (a), DNA methylation levels (b), and mRNA levels (c) of the peroxisome proliferator activated receptor gamma 1 (PPARG1) gene in intramuscular fat (IMF) and muscle portion of Korean cattle steer longissimus dorsi muscle tissue. (a) DNA methylation assay target area of the PPARG1 gene promoter region and transcription factor binding sites. TFCP2, transcription factor CP2. (b) DNA methylation levels were determined by pyrosequencing of bisulfite-treated DNA. Values are the mean+SE (n = 5). (c) The mRNA levels were determined by real-time PCR and normalized against a housekeeping gene. Muscle portion data were normalized to 1.0. Values are the mean+SE (n = 10). * p<0.05; ** p<0.01. PCR, polymerase chain reaction; SE, standard errors.
Mentions: To determine DNA methylation levels, target regions of PPARG1 (Figure 1a) and FABP4 (Figure 2a) genes were selected from the CpG islands, which were searched using CpG Island Searcher (USC Norris Comprehensive Cancer Center, USA; http://www.uscnorris.com/cpgislands2/cpg.aspx). Transcription factor binding sites (Figures 1a and 2a) were determined using TFSEARCH (Computational Biology Research Center, National Institute of Advanced Industrial Science and Technology, Japan; http://www.cbrc.jp/research/db/TFSEARCH.html). We tried to find promoter regions of the genes in which important transcription factor binding sites were located that may regulate gene expression.

Bottom Line: DNA methylation levels of all five CpG sites from the FABP4 gene promoter region were also lower (p<0.001) in the IMF than in the muscle portion.Thus, mRNA levels of both PPARG1 and FABP4 genes were inversely correlated with DNA methylation levels in regulatory regions of CpG sites of the corresponding gene.Our findings suggest that DNA methylation status regulates tissue-specific expression of adipogenic and lipogenic genes in the IMF and muscle portions of LM tissue in Korean cattle.

View Article: PubMed Central - PubMed

Affiliation: Department of Agricultural Biotechnology and Research Institute of Agriculture and Life Sciences, College of Agriculture and Life Sciences, Seoul National University, Seoul 151-921, Korea ; Institute of Green Bio Science Technology, Pyeungchang 232-916, Korea .

ABSTRACT
Epigenetic factors, such as DNA methylation status, may regulate adipogenesis and lipogenesis, thus affecting intramuscular fat (IMF) deposition in longissimus dorsi muscle (LM) of beef cattle. In Korean cattle steers, the LM consists mainly of muscle tissue. However, the LM tissue also contains IMF. We compared the gene expression levels between the IMF and muscle portions of the LM after tissue separation. Real-time polymerase chain reaction analysis showed that the mRNA levels of both adipogenic peroxisome proliferator-activated receptor gamma isoform 1 (PPARG1) and lipogenic fatty acid binding protein 4 (FABP4) were higher (p<0.01) in the IMF than in the muscle portion of the LM. We determined DNA methylation levels of regulatory regions of the PPARG1 and FABP4 genes by pyrosequencing of genomic DNA. DNA methylation levels of two of three CpG sites in the PPARG1 gene promoter region were lower (p<0.05) in the IMF than in the muscle portion of the LM. DNA methylation levels of all five CpG sites from the FABP4 gene promoter region were also lower (p<0.001) in the IMF than in the muscle portion. Thus, mRNA levels of both PPARG1 and FABP4 genes were inversely correlated with DNA methylation levels in regulatory regions of CpG sites of the corresponding gene. Our findings suggest that DNA methylation status regulates tissue-specific expression of adipogenic and lipogenic genes in the IMF and muscle portions of LM tissue in Korean cattle.

No MeSH data available.


Related in: MedlinePlus