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Microarray Analysis of the Gene Expression Profile and Lipid Metabolism in Fat-1 Transgenic Cattle.

Liu X, Bai C, Ding X, Wei Z, Guo H, Li G - PLoS ONE (2015)

Bottom Line: This analysis also identified 11 significantly enriched genes that were involved in the peroxisome proliferator-activated receptor signaling pathway.The information obtained in this study indicated that the introduction of an exogenous fat-1 gene into cattle affects the gene expression profile and the process of lipid metabolism in these animals.These results may provide important insights into how an exogenous fat-1 gene synthesizes n-3 PUFAs in transgenic cattle and other mammals.

View Article: PubMed Central - PubMed

Affiliation: The Key Laboratory of Mammalian Reproductive Biology and Biotechnology of the Ministry of Education, Inner Mongolia University, Hohhot, China; College of Animal Science and Animal Medicine, Tianjin Agriculture University, Tianjin, China.

ABSTRACT
Long-chain n-3 polyunsaturated fatty acids (n-3 PUFAs) are beneficial for human health. However, humans and mammals are unable to synthesize n-3 PUFAs because they lack the n-3 desaturase gene fat-1 and must therefore obtain this type of fatty acid through their diet. Through the production of fat-1 transgenic animals, it is possible to obtain animal products that are rich in n-3 PUFAs, such as meat and milk. The aim of this study was to analyze the gene expression profile and the mechanism of lipid metabolism in fat-1 transgenic cattle and to accumulate important basic data that are required to obtain more efficient fat-1 transgenic cattle. Transcriptome profiling of fat-1 transgenic and wild-type cattle identified differentially expressed genes that are involved in 90 biological pathways, eight pathways of which were related to lipid metabolism processes 36 genes of which were related to lipid metabolism. This analysis also identified 11 significantly enriched genes that were involved in the peroxisome proliferator-activated receptor signaling pathway. These findings were verified by quantitative polymerase chain reaction. The information obtained in this study indicated that the introduction of an exogenous fat-1 gene into cattle affects the gene expression profile and the process of lipid metabolism in these animals. These results may provide important insights into how an exogenous fat-1 gene synthesizes n-3 PUFAs in transgenic cattle and other mammals.

No MeSH data available.


Related in: MedlinePlus

Validation of sixteen microarray differentially expressed genes by RT-qPCR.The fold-change value is expressed as positive when the genes are highly expressed in transgenic cattle and as negative when the genes are highly expressed in wild-type cattle. The gene names are displayed below the histogram.
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pone.0138874.g004: Validation of sixteen microarray differentially expressed genes by RT-qPCR.The fold-change value is expressed as positive when the genes are highly expressed in transgenic cattle and as negative when the genes are highly expressed in wild-type cattle. The gene names are displayed below the histogram.

Mentions: To confirm the gene expression patterns obtained by the microarray analysis, 16 genes were examined by RT-qPCR. We first analyzed 6 of the 11 differentially expressed genes in the PPAR pathway (ACOX1, CPT1B, APOA1, FABP3, FABP2, LPL). The results showed that four genes (ACOX1, CPT1B, FABP2, LPL) were in agreement with the microarray data, with the exception of the APOA1 and FABP3 genes (Fig 4). The APOA1 gene encodes apolipoprotein AI (apo AI), which is the major protein component of HDL and plays a key role in lipid metabolism and transport [34]. Despite the discrepancy observed in our study regarding the expression of APOA1, our RT-qPCR results were consistent with the results of previous studies showing that the levels of apolipoprotein increases in the body through the consumption of high n-3 PUFA-containing foods [35]. FABP3 is a small cytoplasmic protein, and it is involved in fatty acid metabolism because it can transport fatty acids into the mitochondria from the cell membrane. This protein is proposed to be involved in early myocardial development and adult myocardial tissue repair and is thought to be responsible for the modulation of cell growth and proliferation [36]. Our RT-qPCR results also demonstrated the high expression of FABP3 in the fat-1 transgenic cattle, suggesting that the fat-1 gene potentially plays a role in the regulation of the expression of the FABP3 genes. We then analyzed 10 of the 32 differentially expressed genes related to lipid metabolism as obtained by GO enrichment analysis (CYP51A1, LOC782922, MSMO1, HMGCS1, PRODH, ALOX12B, AGMO, FASN, EDN1, PTDSS2). Except for EDN1, both the microarray results and the RT-qPCR data showed that CYP51A1, LOC782922, MSMO1, HMGCS1, and PRODH were up-regulated and that ALOX12B, AGMO, FASN, and PTDSS2 were down-regulated in fat-1 transgenic cattle (Fig 4). EDN1 is a potent vasoconstrictor. The beneficial and detrimental physiological roles of EDN1 were reported in different studies. One study reported that high concentrations of EDN1 might lead to the constriction of coronary arteries, thereby impairing the contractility of the heart, resulting in a dilated cardiomyopathy (DCM) phenotype [37]. However, the present study reported that decreasing the expression of EDN1 by as little as 35% caused severely dilated cardiomyopathy, while a threefold increase in the expression of EDN1 only caused slight cardiac hypertrophy, suggesting that cardiac function was sensitive to even modest changes in EDN1 levels [38]. Our RT-qPCR result indicated that the expression of EDN1 was slightly elevated in fat-1 transgenic cattle, which was inconsistent with our microarray results. However, we speculated that the expression increase of EDN1 may be modest and beneficial for the health of the transgenic cattle, especially for maintaining the normal contractile functions of the heart and blood vessels.


Microarray Analysis of the Gene Expression Profile and Lipid Metabolism in Fat-1 Transgenic Cattle.

Liu X, Bai C, Ding X, Wei Z, Guo H, Li G - PLoS ONE (2015)

Validation of sixteen microarray differentially expressed genes by RT-qPCR.The fold-change value is expressed as positive when the genes are highly expressed in transgenic cattle and as negative when the genes are highly expressed in wild-type cattle. The gene names are displayed below the histogram.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4591129&req=5

pone.0138874.g004: Validation of sixteen microarray differentially expressed genes by RT-qPCR.The fold-change value is expressed as positive when the genes are highly expressed in transgenic cattle and as negative when the genes are highly expressed in wild-type cattle. The gene names are displayed below the histogram.
Mentions: To confirm the gene expression patterns obtained by the microarray analysis, 16 genes were examined by RT-qPCR. We first analyzed 6 of the 11 differentially expressed genes in the PPAR pathway (ACOX1, CPT1B, APOA1, FABP3, FABP2, LPL). The results showed that four genes (ACOX1, CPT1B, FABP2, LPL) were in agreement with the microarray data, with the exception of the APOA1 and FABP3 genes (Fig 4). The APOA1 gene encodes apolipoprotein AI (apo AI), which is the major protein component of HDL and plays a key role in lipid metabolism and transport [34]. Despite the discrepancy observed in our study regarding the expression of APOA1, our RT-qPCR results were consistent with the results of previous studies showing that the levels of apolipoprotein increases in the body through the consumption of high n-3 PUFA-containing foods [35]. FABP3 is a small cytoplasmic protein, and it is involved in fatty acid metabolism because it can transport fatty acids into the mitochondria from the cell membrane. This protein is proposed to be involved in early myocardial development and adult myocardial tissue repair and is thought to be responsible for the modulation of cell growth and proliferation [36]. Our RT-qPCR results also demonstrated the high expression of FABP3 in the fat-1 transgenic cattle, suggesting that the fat-1 gene potentially plays a role in the regulation of the expression of the FABP3 genes. We then analyzed 10 of the 32 differentially expressed genes related to lipid metabolism as obtained by GO enrichment analysis (CYP51A1, LOC782922, MSMO1, HMGCS1, PRODH, ALOX12B, AGMO, FASN, EDN1, PTDSS2). Except for EDN1, both the microarray results and the RT-qPCR data showed that CYP51A1, LOC782922, MSMO1, HMGCS1, and PRODH were up-regulated and that ALOX12B, AGMO, FASN, and PTDSS2 were down-regulated in fat-1 transgenic cattle (Fig 4). EDN1 is a potent vasoconstrictor. The beneficial and detrimental physiological roles of EDN1 were reported in different studies. One study reported that high concentrations of EDN1 might lead to the constriction of coronary arteries, thereby impairing the contractility of the heart, resulting in a dilated cardiomyopathy (DCM) phenotype [37]. However, the present study reported that decreasing the expression of EDN1 by as little as 35% caused severely dilated cardiomyopathy, while a threefold increase in the expression of EDN1 only caused slight cardiac hypertrophy, suggesting that cardiac function was sensitive to even modest changes in EDN1 levels [38]. Our RT-qPCR result indicated that the expression of EDN1 was slightly elevated in fat-1 transgenic cattle, which was inconsistent with our microarray results. However, we speculated that the expression increase of EDN1 may be modest and beneficial for the health of the transgenic cattle, especially for maintaining the normal contractile functions of the heart and blood vessels.

Bottom Line: This analysis also identified 11 significantly enriched genes that were involved in the peroxisome proliferator-activated receptor signaling pathway.The information obtained in this study indicated that the introduction of an exogenous fat-1 gene into cattle affects the gene expression profile and the process of lipid metabolism in these animals.These results may provide important insights into how an exogenous fat-1 gene synthesizes n-3 PUFAs in transgenic cattle and other mammals.

View Article: PubMed Central - PubMed

Affiliation: The Key Laboratory of Mammalian Reproductive Biology and Biotechnology of the Ministry of Education, Inner Mongolia University, Hohhot, China; College of Animal Science and Animal Medicine, Tianjin Agriculture University, Tianjin, China.

ABSTRACT
Long-chain n-3 polyunsaturated fatty acids (n-3 PUFAs) are beneficial for human health. However, humans and mammals are unable to synthesize n-3 PUFAs because they lack the n-3 desaturase gene fat-1 and must therefore obtain this type of fatty acid through their diet. Through the production of fat-1 transgenic animals, it is possible to obtain animal products that are rich in n-3 PUFAs, such as meat and milk. The aim of this study was to analyze the gene expression profile and the mechanism of lipid metabolism in fat-1 transgenic cattle and to accumulate important basic data that are required to obtain more efficient fat-1 transgenic cattle. Transcriptome profiling of fat-1 transgenic and wild-type cattle identified differentially expressed genes that are involved in 90 biological pathways, eight pathways of which were related to lipid metabolism processes 36 genes of which were related to lipid metabolism. This analysis also identified 11 significantly enriched genes that were involved in the peroxisome proliferator-activated receptor signaling pathway. These findings were verified by quantitative polymerase chain reaction. The information obtained in this study indicated that the introduction of an exogenous fat-1 gene into cattle affects the gene expression profile and the process of lipid metabolism in these animals. These results may provide important insights into how an exogenous fat-1 gene synthesizes n-3 PUFAs in transgenic cattle and other mammals.

No MeSH data available.


Related in: MedlinePlus