Altered retinoic acid metabolism in diabetic mouse kidney identified by 18O isotopic labeling and 2D mass spectrometry

Jonathan Starkey, Yingxin Zhao, Rovshan Sadygov, Sigmund J. Haidacher, Wanda S.LeJeune, Nilay Dey, Bruce A. Luxon, Maureen A. Kane, Joseph L. Napoli, Larry Denner, Ronald Tilton

Research output: Contribution to journalArticle

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Abstract

Background: Numerous metabolic pathways have been implicated in diabetes-induced renal injury, yet few studies have utilized unbiased systems biology approaches for mapping the interconnectivity of diabetes-dysregulated proteins that are involved. We utilized a global, quantitative, differential proteomic approach to identify a novel retinoic acid hub in renal cortical protein networks dysregulated by type 2 diabetes. Methodology/Principal Findings: Total proteins were extracted from renal cortex of control and db/db mice at 20 weeks of age (after 12 weeks of hyperglycemia in the diabetic mice). Following trypsinization, 18O- and 16O-labeled control and diabetic peptides, respectively, were pooled and separated by two dimensional liquid chromatography (strong cation exchange creating 60 fractions further separated by nano-HPLC), followed by peptide identification and quantification using mass spectrometry. Proteomic analysis identified 53 proteins with fold change $1.5 and p#0.05 after Benjamini-Hochberg adjustment (out of 1,806 proteins identified), including alcohol dehydrogenase (ADH) and retinaldehyde dehydrogenase (RALDH1/ALDH1A1). Ingenuity Pathway Analysis identified altered retinoic acid as a key signaling hub that was altered in the diabetic renal cortical proteome. Western blotting and real-time PCR confirmed diabetes-induced upregulation of RALDH1, which was localized by immunofluorescence predominantly to the proximal tubule in the diabetic renal cortex, while PCR confirmed the downregulation of ADH identified with mass spectrometry. Despite increased renal cortical tissue levels of retinol and RALDH1 in db/db versus control mice, all-trans-retinoic acid was significantly decreased in association with a significant decrease in PPARb/d mRNA. Conclusions/Significance: Our results indicate that retinoic acid metabolism is significantly dysregulated in diabetic kidneys, and suggest that a shift in all-trans-retinoic acid metabolism is a novel feature in type 2 diabetic renal disease. Our observations provide novel insights into potential links between altered lipid metabolism and other gene networks controlled by retinoic acid in the diabetic kidney, and demonstrate the utility of using systems biology to gain new insights into diabetic nephropathy.

Original languageEnglish (US)
Article numbere11095
JournalPLoS One
Volume5
Issue number6
DOIs
StatePublished - 2010

Fingerprint

retinoic acid
Tretinoin
Metabolism
Labeling
Mass spectrometry
Mass Spectrometry
kidneys
mass spectrometry
Medical problems
Kidney
metabolism
mice
Alcohol Dehydrogenase
Proteins
diabetes
alcohol dehydrogenase
Systems Biology
proteins
proteomics
Retinaldehyde

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Altered retinoic acid metabolism in diabetic mouse kidney identified by 18O isotopic labeling and 2D mass spectrometry. / Starkey, Jonathan; Zhao, Yingxin; Sadygov, Rovshan; Haidacher, Sigmund J.; S.LeJeune, Wanda; Dey, Nilay; Luxon, Bruce A.; Kane, Maureen A.; Napoli, Joseph L.; Denner, Larry; Tilton, Ronald.

In: PLoS One, Vol. 5, No. 6, e11095, 2010.

Research output: Contribution to journalArticle

Starkey, Jonathan ; Zhao, Yingxin ; Sadygov, Rovshan ; Haidacher, Sigmund J. ; S.LeJeune, Wanda ; Dey, Nilay ; Luxon, Bruce A. ; Kane, Maureen A. ; Napoli, Joseph L. ; Denner, Larry ; Tilton, Ronald. / Altered retinoic acid metabolism in diabetic mouse kidney identified by 18O isotopic labeling and 2D mass spectrometry. In: PLoS One. 2010 ; Vol. 5, No. 6.
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AU - S.LeJeune, Wanda

AU - Dey, Nilay

AU - Luxon, Bruce A.

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AU - Denner, Larry

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