retinol-palmitate and Abnormalities--Drug-Induced

Skeletal changes induced by treatment of pregnant rats with four potent teratogens, busulfan, acetazolamide, vitamin A palmitate, and ketoconazole, were evaluated using Alizarin Red S and Alcian Blue double-staining to investigate the relationship between drug-induced skeletal malformations and cartilaginous changes in the fetuses. Pregnant rats (N = 8/group) were treated once or twice between gestation days (GDs) 10 to 13 with busulfan at doses of 3, 10, or 30 mg/kg; acetazolamide at 200, 400, or 800 mg/kg; vitamin A palmitate at 100,000, 300,000, or 1,000,000 IU/kg; or ketoconazole at doses of 10, 30, or 100 mg/kg. Uterine evaluations and fetal external and skeletal examinations were conducted on GD 20. Marked skeletal abnormalities in ribs and hand/forelimb bones such as absent/ short/bent ribs, fused rib cartilage, absent/fused forepaw phalanx, and misshapen carpal bones were induced at the mid- and high-doses of busulfan and acetazolamide and at the high-dose of vitamin A palmitate and ketoconazole. Increased incidences of discontinuous rib cartilage (DRC) and fused carpal bone (FCB) were observed from the low- or mid-dose in the busulfan and acetazolamide groups, and incidences of FCB were increased from the mid-dose in the vitamin A palmitate and ketoconazole groups. Therefore, DRC and FCB were detected at lower doses than those at which ribs and hand/forelimb malformations were observed in the four potent teratogens.

Topics: Abnormalities, Drug-Induced; Acetazolamide; Animals; Busulfan; Carpal Bones; Cartilage; Diterpenes; Dose-Response Relationship, Drug; Female; Fetal Death; Fetal Development; Fetal Resorption; Fetal Weight; Fetus; Ketoconazole; Pregnancy; Random Allocation; Rats; Retinyl Esters; Ribs; Teratogens; Vitamin A

With a view to examine the effects of defined doses of retinyl palmitate (Vit. A) on limb morphogenesis and their effects at the critical time in mouse embryos, pregnant Swiss Webster albino mice were administered retinyl palmitate (10000 or 15000 IU/kg, i.p.) on different days of pregnancy. Vitamin A in 15000 IU/kg, i.p. dose was most effective as produced malformations in the forelimbs by day 10 in 28.6% mice and in the hindlimbs by day 11 in 20.6% mice. Further, two injections in a day with the lower dose (10000 IU/kg, i.p.) had more teratogenic effects than single 15000 IU/kg, i.p. injection. Two injections of either dose on day 10 resulted in higher embryo absorption.

Topics: Abnormalities, Drug-Induced; Animals; Anticarcinogenic Agents; Diterpenes; Drug Administration Schedule; Embryo Loss; Embryo, Mammalian; Extremities; Female; Gestational Age; Injections, Intraperitoneal; Lower Extremity Deformities, Congenital; Mice; Morphogenesis; Pregnancy; Retinyl Esters; Teratoma; Upper Extremity Deformities, Congenital; Vitamin A

A rat model was used to investigate whether high oral doses of vitamin A lead to fetal malformations and to what extent retinyl esters (RES) are transferred from the mother to the fetuses. Retinol and RES concentrations in plasma behave similarly in rats and humans. When high concentrations of vitamin A are administered, plasma retinol concentrations remain relatively constant, whereas plasma RES increased in parallel with the dose. To achieve an elevation from approximately 150 to > 1525 nmol x L(-1) in the experimental group before mating, female Ibm: RORO (spf) rats were fed a maintenance diet enriched with 15.2 x 10(3) retinol equivalents (RE) x kg(-1) at the start and increased stepwise to 52.5 x 10(3) for a total of 8 mo. A parallel subgroup was maintained to measure progress in experimental rats without interference by blood taking. Rats of the control group received the basal diet analyzed to contain 4.5 x 10(3) RE x kg(-1). Before mating the mean body weights of experimental and control rats were not significantly different. All-trans, 13-cis, 4-oxo-all-trans and 5,6-epoxy-all-trans retinoic acid (RA) concentrations were determined in maternal and fetal plasma. With high vitamin A intake, 4-oxo- and 5,6-epoxy RA concentrations were significantly higher in the fetuses than in their mothers. Although these high intakes of vitamin A by the rat dams resulted in high maternal and fetal plasma concentrations of vitamin A and its metabolites, fetal malformations were not observed. This may be due to the fact that circulating RES are not teratogenic and that after crossing the placental barrier, they are stored mainly in fetal liver.

Topics: Abnormalities, Drug-Induced; Animals; Chemical Phenomena; Chemistry, Physical; Chromatography, High Pressure Liquid; Diterpenes; Esters; Female; Maternal-Fetal Exchange; Pregnancy; Rats; Retinyl Esters; Vitamin A

The vitamin A derivative retinoic acid (RA) is a powerful teratogen which can induce severe craniofacial and limb malformations if administered at certain stages of gestation. In addition this compound has been shown to affect patterning in regenerating systems. A classical example is the induction of supernumerary structures along the proximodistal axis of the regenerating amphibian limb. We have investigated the effect of RA on other regenerating systems, the amphibian lower and upper jaws, both in developing and adult animals. We report here that RA does not induce formation of extra structures either in the lower or in the upper jaw of adult newts under experimental conditions where duplications of the regenerating limb occur. However, RA selectively induces severe malformations in the upper jaw regenerate that resemble those induced in avian and mammalian embryos. Analysis of the expression of the newt retinoic acid receptors RAR alpha and delta in upper and lower jaws showed that RAR alpha was expressed at a significant level in the wound epidermis, but not in blastemal cells, whereas no RAR delta could be detected in the regenerate either by in situ hybridization or by using an anti-RAR delta antibody. Therefore, unlike in the limb, in jaws RAR delta is not up-regulated following amputation, and this difference in expression may be causally related to the different effects induced by RA on jaws and limbs. In order to establish whether retinoids affected regeneration of developing jaws in a similar fashion, their effects were studied in animals whose jaws had been amputated at different developmental stages. Under the experimental conditions used overall growth retardation and head defects were observed in the majority of embryos which had been amputated and treated with retinol palmitate (RP) between stages 26-28 and 38-39. In contrast, patterning of upper jaw regenerates in larvae amputated at stage 26-28 and 38-39. In contrast, patterning of upper jaw regenerates in larvae amputated at stage 45 was not significantly affected by the treatment, although the early phase of regeneration was slower than in controls. The different responses to retinoids of regenerating facial structures in embryos, larvae and adults will be discussed.

Topics: Abnormalities, Drug-Induced; Animals; Diterpenes; Gene Expression; In Situ Hybridization; Jaw; Mandible; Maxilla; Notophthalmus; Receptors, Retinoic Acid; Regeneration; Retinyl Esters; Teratogens; Tretinoin; Vitamin A

To examine the effects of vitamin A administered during the preimplantation period, pregnant C3H mice were exposed to teratogenic doses of the vitamin 60 h after copulation. Fetuses were examined for gross abnormalities on the 18th day of gestation and viability, cell number, mitotic index, and chromosome structure were assessed in 81-h blastocysts to determine whether embryotoxic effects were apparent in the preimplantation embryo. There was a reduction in the fetal weight of 18-day fetuses treated in this manner with 15,000 and 30,000 IU vitamin A (p less than 0.0003 in each case), and doses of 10,000 IU and greater were associated with a significantly higher incidence of gross abnormalities. Malformations included exophthalmos, anophthalmia, microphthalmia, exencephaly, exomphalos, and limb defects. Administration of 30,000 IU vitamin A resulted in resorption and intrauterine death in 70% of cases. There was no indication that vitamin A adversely affected 81-h blastocyst viability, cell number, mitotic index, and chromosome structure. The findings suggest that the teratogenic effects that were noted later in fetal life were the result of an action on the developing fetus of the vitamin at a stage later than 81-h and are consistent with the relative resistance of the preimplantation embryo to toxic injury. Persistence of vitamin A, either in the mother or the embryo, is the most likely explanation for the later expression of toxic injury, which is characteristic of the effects that are noted as a result of exposure to the teratogen during the period of organogenesis.

Topics: Abnormalities, Drug-Induced; Animals; Blastocyst; Diterpenes; Female; Fetal Resorption; Litter Size; Mice; Mice, Inbred C3H; Pregnancy; Retinyl Esters; Teratogens; Vitamin A

Evidence from studies of craniofacial anomalies and the evolutionary transition from reptiles to mammals suggests that the temporomandibular joint (TMJ), bony zygomatic arch, middle ear ossicles and mandibular muscle pattern may form a correlated suite of characters. To test the degree of phenotypic interdependence among these features, mandibular arch defects were analysed in prenatal mice. Retinoic palmitate was administered to pregnant mice on day 8.7 to produce test foetuses with malformations of the mandibular arch. A rating scale was developed for each of the four characters so that numerical values could be assigned to each phenotype encountered. Control animals were used to establish normal phenotypes for each character which were assigned a value of 1. Data from each test age, 16, 18 and 19 days postconception, were pooled and Spearmann rank correlation coefficients between each of the traits were calculated. Coefficients (R) range from a high of 0.87, between the TMJ and zygomatic arch, to a low of 0.67 between the zygomatic arch and the mandibular musculature showing highly significant correlations (P less than 0.0001) among all characters. Therefore, the data suggest that the musculoskeletal features of the mandibular arch are phenotypically interdependent during development.

Topics: Abnormalities, Drug-Induced; Animals; Diterpenes; Ear Ossicles; Genetic Linkage; Mandible; Mice; Musculoskeletal System; Phenotype; Retinyl Esters; Temporomandibular Joint; Vitamin A; Zygoma