Priority Paper The metabolic pathway of visual pigment chromophore formation in Drosophila melanogaster

All-trans (3S)-3-hydroxyretinal is formed from all-trans retinal via (3R)-3-hydroxyretinal in the dark

Authors


  • Correspondence to T. Seki, Division of Health Science, Osaka Kyoiku University, Asahigaoka, Kashiwara, Osaka 582-8582, Japan

  • Fax: +81 729 78 3554.

  • Phone: +81 729 78 3611.

  • E-mail:seki@cc.osaka-kyoiku.ac.jp

  • Abbreviations. DWC, lipid-depleted white corn; YC, yellow corn; lutein A, (3R, 3′R, 6′R)-β,ε-carotene-3,3′-diol; β-cryptoxanthin, β-carotene-3-ol; zeaxanthin, β,β'-carotene-3,3′-diol.

Abstract

Carotenoid-depleted fruit flies, Drosophila melanogaster, were reared on yeast/glucose medium containing lipid-depleted white corn grits and cholesterol. After rearing for more than a year, the yield of flies remained constant and the content of 3-hydroxyretinal in a head was three logarithmic units less than that of normal flies reared on medium containing yellow corn grits. When all-trans retinal was supplied as the sole source of retinoids, the flies formed and accumulated all-trans 3-hydroxyretinal in the dark. To examine the metabolic pathway to produce (3S)-3-hydroxyretinal in Drosophila, all-trans retinal was supplemented for two hours to carotenoid-depleted flies in the dark, and the subsequent changes in the composition of 3-hydroxyretinal enantiomers were analyzed using a chiral column on HPLC. The results indicated initial formation of (3R)-3-hydroxyretinal followed by isomerization into the 3S enantiomer. In another set of experiments, the membrane fraction was obtained from the head homogenate of retinoid-depleted flies and an in vitro assay of 3-hydroxyretinal formation from retinal was performed. The 3-hydroxyretinal produced was the 3R enantiomer, supporting the result obtained from the in vivo experiment whereby (3S)-3-hydroxyretinal is produced from retinal via (3R)-3-hydroxyretinal. Addition of NADPH enhanced 3-hydroxyretinal formation and the presence of carbon monoxide inhibited it, suggesting that hydroxylation at the C3 position of retinal occurred via the monooxygenase activity of cytochrome P-450.

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