This research proposes to develop an LC/MS assay to distinguish, identify, and quantitate endogeneous retinoids in biological tissues. Despite the importance of retinoic acid (RA) in development, differentiation, vision, reproduction, and immune response, no sensitive assay exists capable of measuring endogeneous levels. HPLC with various detection schemes lacks sensitivity, GC/MS requires derivitization and cannot resolve geometric isomers of RA, and LC/MS assays to date have not significantly improved detection limits of RA and/or have not been developed as an analytically vigorous assay. The measurement of endogeneous levels of RA and investigation of the hypothesis that RA will colocalize with enzymes that synthesize RA is of great interest towards the understanding of RA function.
Specific aims are (1) to develop and validate a sensitive, specific assay for RA and its isomers in biological samples, (2) to demonstrate the utility of the assay by applying it to animal tissue studies consisting of (a) the measurement of retinoids in wt and CRBP -/- mice, (b) the assessment of the impact of ethanol intoxication on all-trans RA levels in wt mice, and (c) the investigation of whether IRA concentration colocalizes with the enzymatic apparatus that synthesizes it in mouse brain and established cell lines.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Postdoctoral Individual National Research Service Award (F32)
Project #
1F32DK066924-01
Application #
6739982
Study Section
Special Emphasis Panel (ZRG1-F06 (20))
Program Officer
Hyde, James F
Project Start
2004-04-01
Project End
2007-03-31
Budget Start
2004-04-01
Budget End
2005-03-31
Support Year
1
Fiscal Year
2004
Total Cost
$47,296
Indirect Cost
Name
University of California Berkeley
Department
Nutrition
Type
Schools of Earth Sciences/Natur
DUNS #
124726725
City
Berkeley
State
CA
Country
United States
Zip Code
94704
Kane, Maureen A; Folias, Alexandra E; Pingitore, Attilio et al. (2011) CrbpI modulates glucose homeostasis and pancreas 9-cis-retinoic acid concentrations. Mol Cell Biol 31:3277-85
Kane, Maureen A; Bright, Frank V; Napoli, Joseph L (2011) Binding affinities of CRBPI and CRBPII for 9-cis-retinoids. Biochim Biophys Acta 1810:514-8
Kane, Maureen A; Folias, Alexandra E; Pingitore, Attilio et al. (2010) Identification of 9-cis-retinoic acid as a pancreas-specific autacoid that attenuates glucose-stimulated insulin secretion. Proc Natl Acad Sci U S A 107:21884-9
Kane, Maureen A; Folias, Alexandra E; Wang, Chao et al. (2010) Ethanol elevates physiological all-trans-retinoic acid levels in select loci through altering retinoid metabolism in multiple loci: a potential mechanism of ethanol toxicity. FASEB J 24:823-32
Kane, Maureen A; Folias, Alexandra E; Wang, Chao et al. (2008) Quantitative profiling of endogenous retinoic acid in vivo and in vitro by tandem mass spectrometry. Anal Chem 80:1702-8
Kane, Maureen A; Folias, Alexandra E; Napoli, Joseph L (2008) HPLC/UV quantitation of retinal, retinol, and retinyl esters in serum and tissues. Anal Biochem 378:71-9
Kane, Maureen A; Chen, Na; Sparks, Susan et al. (2005) Quantification of endogenous retinoic acid in limited biological samples by LC/MS/MS. Biochem J 388:363-9