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Research ArticleArticle

Quantitative Polymerase Chain Reaction Analysis of the Mouse Cyp2j Subfamily: Tissue Distribution and Regulation

Joan P. Graves, Artiom Gruzdev, J. Alyce Bradbury, Laura M. DeGraff, Huiling Li, John S. House, Samantha L. Hoopes, Matthew L. Edin and Darryl C. Zeldin
Drug Metabolism and Disposition August 2015, 43 (8) 1169-1180; DOI: https://doi.org/10.1124/dmd.115.064139
Joan P. Graves
Immunity, Inflammation, and Disease Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina
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Artiom Gruzdev
Immunity, Inflammation, and Disease Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina
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J. Alyce Bradbury
Immunity, Inflammation, and Disease Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina
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Laura M. DeGraff
Immunity, Inflammation, and Disease Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina
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Huiling Li
Immunity, Inflammation, and Disease Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina
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John S. House
Immunity, Inflammation, and Disease Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina
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Samantha L. Hoopes
Immunity, Inflammation, and Disease Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina
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Matthew L. Edin
Immunity, Inflammation, and Disease Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina
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Darryl C. Zeldin
Immunity, Inflammation, and Disease Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Research Triangle Park, North Carolina
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Abstract

Members of the cytochrome P450 CYP2J subfamily are expressed in multiple tissues in mice and humans. These enzymes are active in the metabolism of fatty acids to generate bioactive compounds. Herein we report new methods and results for quantitative polymerase chain reaction (qPCR) analysis for the seven genes (Cyp2j5, Cyp2j6, Cyp2j8, Cyp2j9, Cyp2j11, Cyp2j12, and Cyp2j13) of the mouse Cyp2j subfamily. SYBR Green primer sets were developed and compared with commercially available TaqMan primer/probe assays for specificity toward mouse Cyp2j cDNA, and analysis of tissue distribution and regulation of Cyp2j genes. Each TaqMan primer/probe set and SYBR Green primer set were shown to be specific for their intended mouse Cyp2j cDNA. Tissue distribution of the mouse Cyp2j isoforms confirmed similar patterns of expression between the two qPCR methods. Cyp2j5 and Cyp2j13 were highly expressed in male kidneys, and Cyp2j11 was highly expressed in both male and female kidneys. Cyp2j6 was expressed in multiple tissues, with the highest expression in the small intestine and duodenum. Cyp2j8 was detected in various tissues, with highest expression found in the skin. Cyp2j9 was highly expressed in the brain, liver, and lung. Cyp2j12 was predominately expressed in the brain. We also determined the Cyp2j isoform expression in Cyp2j5 knockout mice to determine whether there was compensatory regulation of other Cyp2j isoforms, and we assessed Cyp2j isoform regulation during various inflammatory models, including influenza A, bacterial lipopolysaccharide, house dust mite allergen, and corn pollen. Both qPCR methods detected similar suppression of Cyp2j6 and Cyp2j9 during inflammation in the lung.

Footnotes

    • Received March 10, 2015.
    • Accepted May 20, 2015.
  • This work was funded by the Intramural Research Program of the National Institutes of Health, National Institute of Environmental Health Sciences [Z01 ES025034].

  • dx.doi.org/10.1124/dmd.115.064139.

  • ↵Embedded ImageThis article has supplemental material available at dmd.aspetjournals.org.

  • U.S. Government work not protected by U.S. copyright
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Drug Metabolism and Disposition: 43 (8)
Drug Metabolism and Disposition
Vol. 43, Issue 8
1 Aug 2015
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Research ArticleArticle

qPCR analysis of the mouse Cyp2j isoforms

Joan P. Graves, Artiom Gruzdev, J. Alyce Bradbury, Laura M. DeGraff, Huiling Li, John S. House, Samantha L. Hoopes, Matthew L. Edin and Darryl C. Zeldin
Drug Metabolism and Disposition August 1, 2015, 43 (8) 1169-1180; DOI: https://doi.org/10.1124/dmd.115.064139

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Research ArticleArticle

qPCR analysis of the mouse Cyp2j isoforms

Joan P. Graves, Artiom Gruzdev, J. Alyce Bradbury, Laura M. DeGraff, Huiling Li, John S. House, Samantha L. Hoopes, Matthew L. Edin and Darryl C. Zeldin
Drug Metabolism and Disposition August 1, 2015, 43 (8) 1169-1180; DOI: https://doi.org/10.1124/dmd.115.064139
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