The identification of a 9-
            <i>cis</i>
            retinol dehydrogenase in the mouse embryo reveals a pathway for synthesis of 9-
            <i>cis</i>
            retinoic acid

Romert, Anna; Tuvendal, Paulina; Simon, András; Dencker, Lennart; Eriksson, Ulf

doi:10.1073/pnas.95.8.4404

Cited by 88 publications

(67 citation statements)

References 47 publications

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“…It was found that the rostral spinal cord and hindbrain highly express retinaldehydrogenase 2, the enzyme involved in the conversion of retinol into atRA, as well as the cellular retinoic acid-binding protein (CRABP) (34,43,44), two key players in retinoid signaling. However, subsequent isomerization to 9-cis RA is poorly understood (45). In addition, CYP26, which is responsible for the breakdown of RA and, hence, deactivation of RAR or RXR, was found to be expressed in the brain of early embryos (46)(47)(48).…”

Section: Discussionmentioning

confidence: 99%

Spatiotemporal retinoid-X receptor activation detected in live vertebrate embryos

Luria

Furlow

2004

Proc. Natl. Acad. Sci. U.S.A.

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Most studies on the nuclear retinoid-X receptor (RXR) have focused on its role as a heterodimeric partner but less about its own activation pattern during development and the distribution of potential endogenous ligands. The aim of this study is to visualize the distribution of activated RXR␣ in live transgenic Xenopus laevis embryos across a wide range of developmental stages. We adopted a nuclear receptor-Gal4 fusion͞upstream activation sequence-based reporter system for our assay. Strong activation of the RXR␣ ligand-binding domain was observed in a segment of the spinal cord just posterior to the hindbrain. This activation is first detected in neurula stage embryos and persists up to swimming tadpole stages, after which activation strongly declines. Addition of exogenous ligands, such as 9-cis retinoic acid or all-trans retinoic acid, expands the activation of RXR throughout the spinal cord but not in the brain, whereas the RXR-specific ligand LG268 expanded the Gal4 -RXR activation into the brain and olfactory epithelia. Treatment with the RAR-specific ligand 4-(E-2-(5,6,7,8-tetrahydro-5,5,8,8-tetramethyl-2-naphthalenyl)-1-propenyl)benzoic acid or thyroid hormone had no effect on Gal4 -RXR activation, whereas these compounds significantly increased their corresponding Gal4͞ receptor fusion proteins under similar conditions. Embryos expressing a Gal4 -RXR fusion protein with a deletion in the liganddependent activation domain (AF2) show no reporter gene activation. The results shown in this paper reveal a specific activation pattern for Gal4 -RXR␣ specifically in the developing spinal cord and suggest the existence of RXR ligand ''hot-spots'' in this region.Gal4 ͉ transgenesis ͉ Xenopus laevis ͉ nuclear receptor

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Section: Discussionmentioning

confidence: 99%

Spatiotemporal retinoid-X receptor activation detected in live vertebrate embryos

Luria

Furlow

2004

Proc. Natl. Acad. Sci. U.S.A.

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“…25,26 In RPE, where 11-cis-retinoids are relatively abundant, it appears unlikely that this enzyme could play a major role in the production of 9-cis-retinoic acid unless the reaction occurs in a defined intracellular compartment.…”

Section: High-performance Liquid Chromatography Assaymentioning

confidence: 99%

[23] Phase partition and high-performance liquid chromatography assays of retinoid dehydrogenases

Saari

Garwin²,

Haeseleer³

et al. 2000

Methods in Enzymology

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IntroductionPhotoisomerization triggers mammalian vision, converting the chromophore of rhodopsin and cone visual pigments, 11-cis-retinal, to all-trans-retinal. Regeneration of 11-cis-retinal for continued vision occurs in neighboring retinal pigment epithelial (RPE) cells and involves several enzymatic reactions and transcellular diffusion of the retinoids. The overall process is called the visual cycle. 1,2 Developments in our understanding of these reactions have been reviewed. 3-6 At least two different retinoid dehydrogenases play important roles. In photoreceptor cells, an all-trans-specific retinol dehydrogenase catalyzes the reduction of alltrans-retinal by NADPH. 7-9 This reaction is the ultimate step in quenching the reactivity of photoactivated rhodopsin 10 and its rate in mouse retina is equal to the rate of appearance of 11-cis-retinal in rhodopsin. 11,12 In RPE cells a cis-specific retinol dehydrogenase catalyzes the oxidation of 11-cis-retinol to 11-cis-retinal by NAD/NADP. 8,13,14 Both dehydrogenases are members of the short-chain dehydrogenase/reductase superfamily of oxidoreductases. 7, 14 Additional cis-and trans-specific dehydrogenases may be involved in retinoid metabolism in RPE cells 15 (see discussion below).Spectroscopic methods for assaying pyridine nucleotide-dependent dehydrogenases rely on the change in absorbance or fluorescence of the coenzyme. These methods have not been useful with retinoid dehydrogenases because of the relatively low extinction coefficient of the reduced pyridine nucleotides, the turbidity of membrane-associated enzymes, and the fluorescence of retinols and retinyl esters. We developed a phase partition assay for retinoid dehydrogenases that is rapid, accurate, and sensitive. 16 The assay relies on the transfer of tritium from watersoluble [ 3 H]NADPH or [ 3 H]NADH to water-insoluble retinoids. Thus, the activity can be detected by measuring the amount of tritium appearing in a hexane extract of the reaction mixture. Alternatively, the reaction can be followed with [15-3 H]retinoid substrate by measuring the amount of tritium that appears in the aqueous phase as [ 3 H]NADPH or [ 3 H] NADH after hexane extraction. These methods are suitable for dehydrogenase reactions involving water-insoluble reactants and products, such as retinoid, steroid, and eicosanoid dehydrogenases.In this chapter we present methods for the assay of retinoid dehydrogenases by phase partition and high-performance liquid chromatography (HPLC). Examples of the utility and limitation of these assays are provided from studies of retinoid metabolism in the visual cycle. In principle, dehydrogenases that interconvert alcohols and carbonyls can be assayed either in the direction of reduction or oxidation. Pyridine nucleotide-dependent dehydrogenase reactions are pH dependent (see reaction below): low pH favors reduction and high pH favors oxidation. RH 2 + NAD(P) + ⇌ R + NAD(P)H + H +In general, we have used pH 5.5 for phase partition assays that depend on the transfer of tritium to the hydropho...

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“…Production-The RDH5 gene product has been implicated in the production of 9-cis-retinal before this aldehyde is further oxidized to 9-cis-retinoic acids (12,14). However, disabling mutations in men and mice produce no embryonic developmental abnormalities (16)(17)(18)(19).…”

Section: Involvement Of Rdh11-14 In Retinoid Transformation and All-tmentioning

confidence: 99%

“…An enzyme encoded by the RDH5 gene (10,11), 11-cis-RDH, was proposed to be responsible for both 11-cis-retinal and 9-cis-retinal production due to its relaxed substrate specificity (12)(13)(14)(15). However, the disruption of this gene in a mouse model led to uninterrupted production of 11-cis-retinal (16,17) and a lack of any embryonic abnormalities.…”

mentioning

confidence: 99%

Dual-substrate Specificity Short Chain Retinol Dehydrogenases from the Vertebrate Retina

Haeseleer¹,

Jang²,

Imanishi³

et al. 2002

Journal of Biological Chemistry

184

227

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Retinoids are chromophores involved in vision, transcriptional regulation, and cellular differentiation. Members of the short chain alcohol dehydrogenase/reductase superfamily catalyze the transformation of retinol to retinal. Here, we describe the identification and properties of three enzymes from a novel subfamily of four retinol dehydrogenases (RDH11-14) that display dualsubstrate specificity, uniquely metabolizing all-trans-and cis-retinols with C 15 pro-R specificity. RDH11-14 could be involved in the first step of all-trans-and 9-cis-retinoic acid production in many tissues. RDH11-14 fill the gap in our understanding of 11-cis-retinal and all-trans-retinal transformations in photoreceptor (RDH12) and retinal pigment epithelial cells (RDH11). The dualsubstrate specificity of RDH11 explains the minor phenotype associated with mutations in 11-cisretinol dehydrogenase (RDH5) causing fundus albipunctatus in humans and engineered mice lacking RDH5. Furthermore, photoreceptor RDH12 could be involved in the production of 11-cis-retinal from 11-cis-retinol during regeneration of the cone visual pigments. These newly identified enzymes add new elements to important retinoid metabolic pathways that have not been explained by previous genetic and biochemical studies.Retinoids are indispensable light-sensitive elements of vision and also serve as essential modulators of cellular differentiation and proliferation in diverse cell types, including those comprising the epithelium and immune system. Retinoids modulate the growth of both normal and malignant cells through their binding to retinoid receptors. All-trans-retinoic acid signals through specific interactions with the nuclear retinoic acid receptors, whereas its isomer, 9-cis-retinoic acid, is a high affinity ligand of retinoic acid receptors and retinoid X receptors. In the retina, light-dependent photoisomerization of 11-cis-retinylidene to the all-transretinylidene moiety of rod and cone photoreceptors is a key reaction that triggers visual * This research was supported by National Institutes of Health Grants EY08061, CA75173, and DK59125, a grant from Research to Prevent Blindness, Inc. (to the Department of Ophthalmology, University of Washington), and by the E. K. Bishop Foundation. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. S The on-line version of this article (available at http://www.jbc.org) contains Supplemental Figs. 1 and 2 and Tables 1-3. § These authors contributed equally to this work. § § Recipient of a Research to Prevent Blindness Senior Investigator Award. To whom correspondence should be addressed: Dept. of Ophthalmology, University of Washington, Seattle,; E-mail: palczews@u.washington.edu. Transformations of retinoids occur mostly through enzymatic or photochemical reactions, although they readily isomerize non-enzymatically to thermodynamic equilibriu...

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The identification of a 9- cis retinol dehydrogenase in the mouse embryo reveals a pathway for synthesis of 9- cis retinoic acid

Cited by 88 publications

References 47 publications

Spatiotemporal retinoid-X receptor activation detected in live vertebrate embryos

Spatiotemporal retinoid-X receptor activation detected in live vertebrate embryos

[23] Phase partition and high-performance liquid chromatography assays of retinoid dehydrogenases

Dual-substrate Specificity Short Chain Retinol Dehydrogenases from the Vertebrate Retina

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