Abstract
Oral administration of retinol (50 mg/kg) to NMRI mice on day 11 of gestation (vaginal plug = day 0) led to the metabolic formation of high quantities of all-trans retinoic acid and all-trans-4-oxoretinoic acid, both known as potent teratogenic agents in the mouse. A 96% reduction of the area under the concentration-versus-timecurve (AUC) of metabolically generated alltrans retinoic acid in maternal plasma, and an 84% decrease in the embryonic AUC were observed when mice had been pretreated with the alcohol dehydrogenase inhibitor 4-methylpyrazole. A similar reduction was observed for the major metabolite of all-trans retinoic acid in the mouse, all-trans-4-oxoretinoic acid. However, 4-methylpyrazole pretreatment decreased the AUC of retinol by 10% in maternal plasma and 15% in embryo. Treatment with retinol alone resulted in 55.6%, 43.9% and 56.0% skeletal anomalies of the forelimbs, hindlimbs and craniofacial structures, respectively. Pretreatment with 4-methylpyrazole lowered the retinol induced skeletal defects to 31.3%, 24.0% and 31.3%, respectively, in the forelimb, hindlimb and craniofacial region. Typical retinoid-induced malformations for gestational day 11, e.g. bent or reduced zeugopod or stylopod elements, or cleft palate, were significantly reduced by 4-methylpyrazole pretreatment but were still detected in significantly higher prevalence than in control mice. These data suggest that the teratogenic activity of a single high dose of vitamin A in mouse is partially but not exclusively dependent on the metabolic activation of retinol to all -trans retinoic acid. Thus it could be hypothesized that retinol is either a proximate teratogen or a coteratogen with all -trans retinoic acid.
Similar content being viewed by others
References
Algar EM, Seeley T-L, Holmes RS (1983) Purification and molecular properties of mouse alcohol dehydrogenase isozymes. Eur J Biochem 137: 139–147
Burnell JC, Bosron WF (1989) Genetic polymorphism of human liver alcohol dehydrogenase and kinetic properties of the isoenzymes. In: Crow KE, Batt RD (eds) Human metabolism of alcohol, vol II. CRC Press, Boca Raton, FL, pp 65–75
Connor MJ, Smit MH (1987) Terminal-group oxidation of retinol by mouse epidermis. Inhibition in vitro and in vivo. Biochem J 244: 489–492
Creech Kraft J, Löfberg B, Chahoud I, Bochert G, Nau H (1989) Teratogenicity and placental transfer of all-trans, 13-cis, 4-oxo-all-trans, and 4-oxo-13-cis-retinoic acid after administration of a low oral dose during organogenesis in mice. Toxicol Appl Pharmacol 100: 162–176
Dawson AA (1926) A note on the staining of the skeleton of cleared specimen with alizarin red S. Stain Technol 1: 124–124
Dollé P, Ruberte E, Kastner P, Petkovich M, Stoner CM, Gudas L, Chambon P (1989) Differential expression of the genes encoding the retinoic acid receptors α, β, γ, and CRABP in the developing limbs of the mouse. Nature 342: 702–705
Duester G (1991) A hypothetical mechanism for fetal alcohol syndrome involving ethanol inhibition of retinoic acid synthesis at the alcohol dehydrogenase step. Alcohol Clin Exp Res 15: 568–572
Duester G, Shean ML, McBride MS, Stewart MJ (1991) Retinoic acid response element in the human alcohol dehydrogenase gene ADH3: implications for regulation of retinoic acid synthesis. Mol Cell Biol 11: 1638–1646
Durston AJ, Timmermans JPM, Hage WJ, Hendricks HFJ, de Vries NJ, Heideveld M, Nieuwkoop PD (1989) Retinoic acid causes an anteroposterior transformation in the developing central nervous system. Nature 340: 140–144
Eckhoff C, Löfberg B, Chahoud I, Bochert G, Nau H (1989) Transplacental pharmacokinetics and teratogenicity of a single dose of retinol (vitamin A) during organogenesis in the mouse. Toxicol Lett 48: 171–184
Eckhoff C, Wittfoht W, Nau H, Slikker W (1990) Characterization of oxidized and glucuronidated metabolites of retinol in monkey plasma by thermospray liquid chromatography/mass spectrometry. Biomed Environ Mass Spectrom 19: 428–433
Geelen JAG (1979) Hypervitaminosis A induced teratogenesis. CRC Crit Rev Toxicol 6: 351–376
Giguère V, Ong ES, Evans RM, Tabin CJ (1989) Spatial and temporal expression of the retinoic acid receptor in the regenerating amphibian limb. Nature 337: 566–569
Holmes RS (1978) Electrophoretic analyses of alcohol dehydrogenase, aldehyde dehydrogenase, aldehyde oxidase, sorbitol dehydrogenase and xanthine oxidase from mouse tissues. Comp Biochem Physiol B 61:339–346
Jetten AM, Jetten MER (1979) Possible role of retinoic acid binding protein in retinoid stimulation of embryonic carcinoma cell differentiation. Nature 278: 180–182
Julia P, Farres J, Pares X (1987) Characterization of three isoenzymes of rat alcohol dehydrogenase. Tissue distribution and physical and enzymatic properties. Eur J Biochem 162: 179–189
Kochhar DM, Penner JD, Satre MA (1988) Derivation of retinoic acid and metabolites from a teratogenic dose of retinol (vitamin A) in mice. Toxicol Appl Pharmacol 96: 429–441
Leo MA, Kim C-I, Lieber CS (1987) NAD+-dependent retinol dehydrogenase in liver microsomes. Arch Biochem Biophys 259: 241–249
Li T-K, Theorell H (1969) Human liver alcohol dehydrogenase: inhibition by pyrazole and pyrazole analogs. Acta Chem Scand 23: 892
Löfberg B, Chahoud I, Bochert G, Nau H (1990a) Teratogenicity of the 13-cis and all-trans-isomers of the aromatic retinoid etretin: Correlation to transplacental pharmacokinetics in mice during organogenesis after a single oral dose. Teratology 41: 707–716
Löfberg B, Reiners J, Spielmann H, Nau H (1990b) Teratogenicity of steady-state concentrations of etretinate and metabolite acitretin maintained in maternal plasma and embryo by intragastric infusion during organogenesis in the mouse: a possible model for the extended elimination phase in human therapy. Dev Pharmacol Ther 15: 45–51
MacDonald PN, Ong DE (1988) Evidence for a lecithin: retinol acyltransferase activity in the rat small intestine. J Biol Chem 263: 12478–12482
Maden M, Ong DE, Summerbell D, Chytil F (1988) Spatial distribution of cellular protein binding to retinoic acid in the chick limb bud. Nature 335: 733–735
Mezey E, Holt PR (1971) The inhibitory effect of ethanol on retinol oxidation by human liver and cattle retina. Exp Mol Pathol 15: 148–156
Napoli JL, Race KR (1987) The biosynthesis of retinoic acid from retinol by rat tissues in vitro. Arch Biochem Biophys 255: 95–101
Napoli JL, Race KR (1990) Microsomes convert retinol and retinal into retinoic acid and interfere in the conversions catalyzed by cytosol. Biochim Biophys Acta 1034: 228–232
Napoli JL, Posch KP, Fiorella PD, Boermann MHEM (1991) Physiological occurrence, biosynthesis and metabolism of retinoic acid: evidence for roles of cellular retinol-binding protein (CRBP) and cellular retinoic acid-binding protein (CRABP) in the pathway of retinoic acid homeostasis. Biomed Pharmacother 45: 131–143
Nau H (1990) Correlation of transplacental and maternal pharmacokinetics of retinoids during organogenesis with teratogenicity. Methods Enzymol 190: 437–448
Nau H, Creech Kraft J, Eckhoff C, Löfberg B (1989) Interpretation of retinoid teratogenesis by transplacental pharmacokinetics. In: Reichert U, Shroot B (eds) Pharmacology of retinoids in the skin, vol II. Karger, Basel, pp 165–173
Ong DE (1984) A novel retinol binding protein from rat: purification and partial characterization. J Biol Chem 259: 1476–1482
Pearce JC, Jelly JA, Fernandes KA, Leavens WJ, McDowall RD (1986) Analysis of 5-(4-acetamindophenyl)pyrazin-2(1H)-one (SK&F 94120) in plasma with an Analytichem automated sample processor liquid chromatography module. J Chromatogr 353: 371–378
Posch KC, Enright WJ, Napoli JL (1989) Retinoic acid synthesis by cytosol from the alcohol dehydrogenase negative deermouse. Arch Biochem Biophys 274: 171–178
Reiners J, Löfberg B, Creech Kraft J, Kochhar DM, Nau H (1988) Transplacental pharmacokinetics of teratogenic doses of etretinate and other aromatic retinoids in mice. Reprod Toxicol 2: 19–29
Ross AC (1981) Separation of long-chain fatty acid esters of retinol by high-performance liquid chromatography. Anal Biochem 115: 324–330
Shih TW, Hill DL (1991) Conversion of retinol to retinal by rat liver microsomes. Drug Metab Dispos 19: 332–335
Siegenthaler G, Saurat J-H, Ponec M (1990) Retinol and retinal metabolism. Relationship to the state of differentiation of cultured human keratinocytes. Biochem J 268: 371–378
Takahashi YI, Smith JE, Winick M, Goodman DS (1975) Vitamin A deficiency and fetal growth and development in the rat. J Nutr 105: 1299–1310
Takase S, Ong DE, Chytil F (1986) Transfer of retinoic acid from its complex with cellular retinoic acid-binding protein to the nucleus. Arch Biochem Biophys 247: 328
Teratology Society (1987) Teratology Society Position Paper: recommendations for vitamin A use during pregnancy. Teratology 35: 269–275
Thaller C, Eichele G (1987) Identification and spatial distribution of retinoids in the developing chick limb bud. Nature 327: 625–628
Thaller C, Eichele G (1990) Isolation of 3,4-didehydroretinoic acid, a novel morphogenetic signal in the chick wing bud. Nature 345: 815–819
Thompson JN, Howell JM, Pitt GAJ (1964) Vitamin A and reproduction in rats. Proc R Soc Lond B 159: 510–535
Vallee BL, Bazzone TJ (1983) Isozymes of human liver alcohol dehydrogenase. Curr Top Biol Med Res 8: 219–244
Willhite CC, Dawson MI, Williams KJ (1984) Structure-activity relationships of retinoids in developmental toxicology. I. Studies on the nature of the polar terminus of the vitamin A molecule. Toxicol Appl Pharmacol 74: 397–410
Willhite CC, Wier PJ, Berry DL (1989) Dose response and structure-activity considerations in retinoid-induced dysmorphogenesis. Crit Rev Toxicol 20: 113–135
Wilson JG, Roth CB, Warkany J (1953) An analysis of the syndrome of malformations induced by maternal vitamin A deficiency. Effects of restoration of vitamin A at various times during gestation. Am J Anat 92: 189–218
Wolf G (1990) Recent progress in vitamin A research: nuclear retinoic acid receptors and their interaction with gene elements. J Nutr Biochem 1: 284–289
Zachman RD, Olson JA (1961) A comparison of retinene reductase and alcohol dehydrogenase of rat liver. J Biol Chem 236: 2309–2313
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Collins, M.D., Eckhoff, C., Chahoud, I. et al. 4-Methylpyrazole partially ameliorated the teratogenicity of retinol and reduced the metabolic formation of all-trans-retinoic acid in the mouse. Arch Toxicol 66, 652–659 (1992). https://doi.org/10.1007/BF01981505
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01981505