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

doi:10.1016/0041-008X(89)90099-9

Toxicology and Applied Pharmacology

Volume 100, Issue 1, August 1989, Pages 162-176

https://doi.org/10.1016/0041-008X(89)90099-9 Get rights and content

Abstract

13-cis-Retinoic acid (isotretinoin) is teratogenic in humans at therapeutic doses (0.5–1.5 mg/kg) but only marginally teratogenic in the mouse at a high dose of 100 mg/kg. Previous results explained why the cis isomer of retinoic acid was much less teratogenic than the trans isomer in mice. It was found that the placental transfer of all-trans retinoic acid to the mouse embryo was far greater than that of the 13-cis isomer. Since our previous study had been performed with exceedingly high doses (100 mg/kg) of 13-cis-retinoic acid and all-trans-retinoic acid, we have now performed additional experiments with 10-fold lower doses. Studies were also done with the main metabolites of the two retinoids (the 4-oxo-derivatives) to elucidate the metabolism, pharmacokinetics, and teratogenicity of each single compound. It was shown that all-trans-retinoic acid and 4-oxo-all-trans-retinoic acid were extremely teratogenic, whereas their corresponding cis isomers caused only 2% cleft palate. Embryonic exposure to the trans isomers was likewise higher than that to the cis isomers, as shown by the far higher embryonic peak concentrations and by the 30-fold higher areas under the concentration-time curve values reached for the trans isomers compared with the cis isomers. At 8 hr, embryo/maternal plasma ratios were higher than 1 after administration of the all-trans compounds. Concentrations found in the placenta and yolk sac were higher for the trans forms than for the cis forms. We propose a model for a facilitated transport of the all-trans forms to the developing embryo and suggest that the conversion to the trans isomer and trans metabolite could play a major role in the teratogenicity of 13-cis-retinoic acid in humans.

References (31)

W. Bollag
Vitamins and retinoids: From nutrition to pharmacotherapy in dermatology and oncology
Lancet
(1983)
J.T. Braun et al.
Isotretinoin dysmorphic syndrome
Lancet
(1984)
J. Kamm et al.
Preclinical and clinical toxicology of selected retinoids
S. Puhvel et al.
Cellular retinoic acid-binding proteins in human epidermis and sebaceous follicles
J. Invest. Dermatol.
(1984)
F.W. Rosa
Teratogenicity of isotretinoin
Lancet
(1983)
G. Siegenthaler et al.
Retinol-binding protein in human serum: Conformational changes induced by retinoic acid binding
Biochem. Biophys. Res. Commun.
(1987)
P.S. Bernstein et al.
Isomerization of all-trans-retinoids to 11-cis-retinoids in vitro
R.K. Brazzel et al.
Pharmacolinetics of isotretinoin during repetitive dosing to patients
Eur. J. Pharmacol.
(1983)
I. Chahoud et al.
Controlled breeding of laboratory animals
F. Chytil et al.
Cellular vitamin A binding proteins
Vitam. Horm.
(1978)

J. Creech Kraft et al.

Automated determination of 13-cis and all-trans-retinoic acid, their 4-oxo-metabolites and retinol in plasma, amniotic fluid and embryo by reversed-phase high-performance liquid chromatography with a precolumn switching technique

J. Liq. Chromatogr.

(1988)

J. Creech Kraft et al.

Low teratogenicity of 13-cis-retinoic acid (isotretinoin) in the mouse corresponds to low embryo concentration during organogenesis: Comparison to the all-trans isomer

Toxicol. Appl. Pharmacol.

(1987)

A.A. Dawson

A note on the staining of the skeleton of cleared specimen with alizarin red S

Stain Technol.

(1926)

L. Dencker et al.

Saturable accumulation of retinoic acid in neural and neural crest derived cells in early embryonic development

Dev. Pharmacol. Ther.

(1987)

W.D. Faber et al.

13-cis-Retinoic acid (isotretinoin) uptake and metabolism by the human placenta

Teratology

(1988)

Cited by (93)

Administration of retinoic acid to pregnant mice increases the number of fetal mouse glomeruli
2022, Biochemistry and Biophysics Reports
Citation Excerpt :
It is assumed that there is a difference in the transfer of RA to plasma between oral and intraperitoneal administration routes. The plasma half-life of all-trans-RA is short, approximately 30–60 min in the mouse fetus [20]. Retinoid teratogenicity correlates with the concentration-time curve, or area under the curve (AUC) [28].
The prevalence of chronic kidney disease (CKD) is increasing worldwide, and CKD is a serious global health problem. Low glomerular number is one of the risk factors for CKD; therefore, the glomerular number is associated with the risk of CKD. Increasing the glomerular number above normal levels may reduce the risk of CKD. It has been reported that, in vitro, the addition of retinoic acid (RA) to the culture medium increases the glomerular number. However, there is no report of an increase in glomerular number above normal levels with the addition of RA in vivo. In this study, RA (20 mg/kg) was administered intraperitoneally to pregnant mice once at embryonic day (E) 10.5, E12.5, E14.5, or E16.5. The fetuses were harvested at E18.5 and fetal mouse kidneys were evaluated. Fetal kidney volume and weight were significantly increased in the E16.5 group compared to the control group. The total glomerular number in the E16.5 group was also approximately 1.46 times higher than that in the control group. In summary, we established a method to increase the glomerular number in the fetal kidney by administration of RA to pregnant mice at E16.5. These results will facilitate the investigation of whether CKD risk is reduced when the glomerular number increases above normal.
Comparison of osteoblast and cardiomyocyte differentiation in the embryonic stem cell test for predicting embryotoxicity in vivo
2014, Reproductive Toxicology
One of the most studied alternative embryotoxicity assays is the embryonic stem cell test, in which the effect of compounds on cardiomyocyte differentiation is evaluated (subsequently termed the ESTc). This single differentiation endpoint may limit the predictive value of the assay. We recently published a novel embryonic stem cell based osteoblast differentiation assay (subsequently termed the ESTo), in which we studied the effect of six embryotoxic compounds. Differentiation is monitored via the differential expression of three genes related to osteogenesis (Runx2, SPARC and collagen type I). In the current study, we evaluated the effect of 14 additional compounds in the ESTo, to assess its added value as compared to the ESTc. To this end, we compared the effects of the compounds in the ESTo to their effects in the ESTc and to their published in vivo developmental toxicity profiles. The results show that there is a high overall correlation between compound potencies as regards inhibition of osteoblast and cardiomyocyte differentiation. Moreover, the results in both the ESTo and ESTc showed a significant correlation to in vivo developmental toxicity potency ranking of compounds tested. Interestingly, the embryotoxic effect of TCDD could only be detected using the ESTo, which can be explained based on its mechanism of action and its known inhibitory effect on osteogenesis. The results of TCDD suggest that incorporating the ESTo into a testing battery together with the ESTc could improve the overall predictive value of the battery.
A sensitive and specific method for measurement of multiple retinoids in human serum with UHPLC-MS/MS
2012, Journal of Lipid Research
Retinol (vitamin A) circulates at 1–4 μM concentration and is easily measured in serum. However, retinol is biologically inactive. Its metabolite, retinoic acid (RA), is believed to be responsible for biological effects of vitamin A, and hence the measurement of retinol concentrations is of limited value. A UHPLC-MS/MS method using isotope-labeled internal standards was developed and validated for quantitative analysis of endogenous RA isomers and metabolites. The method was used to measure retinoids in serum samples from 20 healthy men. In the fed state, the measured concentrations were 3.1 ± 0.2 nM for atRA, 0.1 ± 0.02 nM for 9-cisRA, 5.3 ± 1.3 nM for 13-cisRA, 0.4 ± 0.4 nM for 9,13-dicisRA, and 17.2 ± 6.8 nM for 4oxo-13-cisRA. The concentrations of the retinoids were not significantly different when measured after an overnight fast (3.0 ± 0.1 nM for atRA, 0.09 ± 0.01nM for 9-cisRA, 3.9 ± 0.2 nM for 13-cisRA, 0.3 ± 0.1 nM for 9,13-dicisRA, and 11.9 ± 1.6 nM for 4oxo-13-cisRA). 11-cisRA and 4OH-RA were not detected in human serum. The high sensitivity of the MS/MS method combined with the UHPLC separation power allowed detection of endogenous 9-cisRA and 4oxo-atRA for the first time in human serum.
Metabolism of Retinol During Mammalian Placental and Embryonic Development
2007, Vitamins and Hormones
Citation Excerpt :
The presence of measurable hepatic vitamin A stored at birth is indicative of the functionality of placental transport during gestation (Ross and Gardner, 1994; Satre et al., 1992). A number of studies have investigated the ability of retinoids to pass through the placental barrier in mice or rabbits (Collins et al., 1994; Creech Kraft et al., 1989, 1991; Kochhar et al., 1988; Sass et al., 1999; Ward and Morriss‐Kay, 1995). It is well established that each retinoid (e.g., retinol, 13‐cis, 9‐cis, all‐trans RA, and their glycuronoconjugates) presents a specific rate of transfer.
Retinol (vitamin A) is a fat‐soluble nutrient indispensable for a harmonious mammalian gestation. The absence or excess of retinol and its active derivatives [i.e., the retinoic acids (RAs)] can lead to abnormal development of embryonic and extraembryonic (placental) structures. The embryo is unable to synthesize the retinol and is strongly dependent on the maternal delivery of retinol itself or precursors: retinyl esters or carotenoids. Before reaching the embryonic tissue, the retinol or the precursors have to pass through the placental structures. During this placental step, a simple diffusion of retinol can occur between maternal and fetal compartments; but retinol can also be used in situ after its activation into RA¹ or stored as retinyl esters. Using retinol‐binding protein knockout model, an alternative way of embryonic retinol supply was described using retinyl esters incorporated into maternal chylomicrons. In the embryo, the principal metabolic event occurring for retinol is its conversion into RAs, the active molecules implicated on the molecular control of embryonic morphogenesis and organogenesis. All these placental and embryonic events of retinol transport and metabolism are highly regulated. Nevertheless, some genetic and/or environmental abnormalities in the transport and/or metabolism of retinol can be related to developmental pathologies during mammalian development.
Retinoic Acid and the Heart
2007, Vitamins and Hormones
Citation Excerpt :
The teratogenic effects of a high dose of vitamin A were first demonstrated in pregnant rats (Cohlan, 1953, 1954). RA was shown to be a more potent teratogen than retinol in several animal models (Creech Kraft et al., 1989; Morriss and Steele, 1977; Shenefelt, 1972a). RA has previously been shown to have teratogenic effects on heart development in mammalian embryos (Fantel et al., 1977; Kalter and Warkany, 1961; Robens, 1970; Shenefelt, 1972b; Taylor et al., 1980).
Retinoic acid (RA), the active derivative of vitamin A, by acting through retinoid receptors, is involved in signal transduction pathways regulating embryonic development, tissue homeostasis, and cellular differentiation and proliferation. RA is important for the development of the heart. The requirement of RA during early cardiovascular morphogenesis has been studied in targeted gene deletion of retinoic acid receptors and in the vitamin A‐deficient avian embryo. The teratogenic effects of high doses of RA on cardiovascular morphogenesis have also been demonstrated in different animal models. Specific cardiovascular targets of retinoid action include effects on the specification of cardiovascular tissues during early development, anteroposterior patterning of the early heart, left/right decisions and cardiac situs, endocardial cushion formation, and in particular, the neural crest. In the postdevelopment period, RA has antigrowth activity in fully differentiated neonatal cardiomyocytes and cardiac fibroblasts. Recent studies have shown that RA has an important role in the cardiac remodeling process in rats with hypertension and following myocardial infarction. This chapter will focus on the role of RA in regulating cardiomyocyte growth and differentiation during embryonic and the postdevelopment period.
Plasma retinoid profile in bullfrogs, Rana catesbeiana, in relation to agricultural intensity of sub-watersheds in the Yamaska River drainage basin, Québec, Canada
2005, Aquatic Toxicology
Amphibian populations are decreasing globally and the causes are presently unclear. Retinoids have been extensively studied in other vertebrate classes where they are associated with pleiotropic effects such as susceptibility to disease (including cancer and parasitic infections), deformities and reproduction. To investigate the hypothesis that retinoid homeostasis is influenced by agricultural activities, blood samples were collected from adult bullfrogs, Rana catesbeiana, at each of six sub-watersheds chosen to represent a gradient of agricultural intensity within the Yamaska River drainage basin. Samples of surface water were collected at each of the study sites approximately 1 month after spraying and analyzed for 53 pesticides. Male body weight was significantly different (p < 0.001) between study sites with the smallest bullfrogs captured from the Rivière à la Barbue sub-watershed associated with high agricultural intensity. A significant linear regression (p < 0.001; R² = 0.176) was obtained between plasma retinol and body weight. Plasma retinol concentrations were significantly different between study sites (p < 0.001) being lowest at both Rivière Noire and Rivière à la Barbue. More than 60% of the land area in these sub-watersheds is under intensive corn–soya cultivation and surface water contained the highest concentrations of the herbicides atrazine, deethyl-atrazine, simazine, metolachlor, dimethenamide, chlopyralide, dicamba and bentazone. Plasma 13-cis-4-oxo-retinoic acid was significantly different (p < 0.001) between sub-watersheds, however this effect was apparently unrelated to agricultural intensity. Plasma retinol was negatively correlated (p = 0.026; r = −0.237) with plasma 13-cis-4-oxo-retinoic acid. These results suggest that retinoid homeostasis in bullfrogs may be influenced by agricultural practices.

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Toxicology and Applied Pharmacology

Abstract

References (31)

Vitamins and retinoids: From nutrition to pharmacotherapy in dermatology and oncology

Lancet

Isotretinoin dysmorphic syndrome

Lancet

Preclinical and clinical toxicology of selected retinoids

Cellular retinoic acid-binding proteins in human epidermis and sebaceous follicles

J. Invest. Dermatol.

Teratogenicity of isotretinoin

Lancet

Retinol-binding protein in human serum: Conformational changes induced by retinoic acid binding

Biochem. Biophys. Res. Commun.

Isomerization of all-trans-retinoids to 11-cis-retinoids in vitro

Pharmacolinetics of isotretinoin during repetitive dosing to patients

Eur. J. Pharmacol.

Controlled breeding of laboratory animals

Cellular vitamin A binding proteins

Vitam. Horm.

Automated determination of 13-cis and all-trans-retinoic acid, their 4-oxo-metabolites and retinol in plasma, amniotic fluid and embryo by reversed-phase high-performance liquid chromatography with a precolumn switching technique

J. Liq. Chromatogr.

Low teratogenicity of 13-cis-retinoic acid (isotretinoin) in the mouse corresponds to low embryo concentration during organogenesis: Comparison to the all-trans isomer

Toxicol. Appl. Pharmacol.

A note on the staining of the skeleton of cleared specimen with alizarin red S

Stain Technol.

Saturable accumulation of retinoic acid in neural and neural crest derived cells in early embryonic development

Dev. Pharmacol. Ther.

13-cis-Retinoic acid (isotretinoin) uptake and metabolism by the human placenta

Teratology

Cited by (93)

Administration of retinoic acid to pregnant mice increases the number of fetal mouse glomeruli

Comparison of osteoblast and cardiomyocyte differentiation in the embryonic stem cell test for predicting embryotoxicity in vivo

A sensitive and specific method for measurement of multiple retinoids in human serum with UHPLC-MS/MS

Metabolism of Retinol During Mammalian Placental and Embryonic Development

Retinoic Acid and the Heart

Plasma retinoid profile in bullfrogs, Rana catesbeiana, in relation to agricultural intensity of sub-watersheds in the Yamaska River drainage basin, Québec, Canada