The long-term goals of my research program include defining the biochemical, molecular and endocrine aspects of RA (retinoic acid) biosynthesis. This project will focus on the second step in the pathway of RA biogenesis from retinol (vitamin A), catalyzed by RALDH1-4 (retinal dehydrogenase types 1-4), and will test the umbrella hypothesis that each RALDH makes a unique and essential contribution to RA biogenesis.
Specific Aims 1 would determine the contribution of RALDH 1, 2 and 3 to RA biosynthesis in the brain, focusing on the hippocampus.
This aim would test the hypotheses that: hippocampus neurons biosynthesize RA from retinol; BDNF enhances neuron conversion of retinol into RA; BDNF functions by increasing RALDH3 mRNA and protein; and exogenous retinoids alter RA biosynthesis in the brain.
This aim will include quantifying RA concentrations in specific brain areas by LC/MS/MS (lower limit of detection of 10 fmol) during aging and treatment with exogenous retinoids.
Specific Aims 2 -4 will: 2) generate and evaluate RALDH4-null mice; 3) identify the RALDH 1 gene regulatory regions that mediate tissue-specific responses to retinoids in vivo; 4) generate and evaluate conditional RALDH2 knockouts. Each of these aims represents a logical progression in evaluating the specific contribution of an RALDH or RALDHs to RA biosynthesis.
Each aim will rely on integrating molecular genetics approaches (transgenics and knockouts) with analytical techniques (hplc, LC/MS) and applying both to metabolic studies in vitro and in the whole animal. The fundamental insight and reagents generated will be useful to identify diseases and age-related degeneration that may be caused or exacerbated by impaired RA biosynthesis.

Agency
National Institute of Health (NIH)
Institute
National Institute on Aging (NIA)
Type
Research Project (R01)
Project #
2R01AG013566-11
Application #
6966516
Study Section
Integrative Nutrition and Metabolic Processes Study Section (INMP)
Program Officer
Finkelstein, David B
Project Start
1995-07-10
Project End
2010-06-30
Budget Start
2005-09-01
Budget End
2006-06-30
Support Year
11
Fiscal Year
2005
Total Cost
$297,494
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
Napoli, Joseph L (2012) Physiological insights into all-trans-retinoic acid biosynthesis. Biochim Biophys Acta 1821:152-67
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; 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
Chrispell, Jared D; Feathers, Kecia L; Kane, Maureen A et al. (2009) Rdh12 activity and effects on retinoid processing in the murine retina. J Biol Chem 284:21468-77
Chen, Na; Onisko, Bruce; Napoli, Joseph L (2008) The nuclear transcription factor RARalpha associates with neuronal RNA granules and suppresses translation. J Biol Chem 283:20841-7
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
Chen, Na; Napoli, Joseph L (2008) All-trans-retinoic acid stimulates translation and induces spine formation in hippocampal neurons through a membrane-associated RARalpha. FASEB J 22:236-45
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
McCaffery, Peter; Koul, Omanand; Smith, Deborah et al. (2004) Ethanol increases retinoic acid production in cerebellar astrocytes and in cerebellum. Brain Res Dev Brain Res 153:233-41

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