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Behavioral teratogenesis: an extension to the teratogenesis of functions.

Major malformations correspond to pathology during the first 2 months of gestation. Thereafter, histological and biochemical abnormalities can result from different negative maternal incidents and, without obvious malformations, change the phenotype of the conceptus. These abnormalities lead to essentially functional disorders often compatible with life and to more or less serious handicaps. All systems, when they arrive at such a stage of development, can be theoretically involved. The central nervous system (CNS) offers a very clear example of such weaknesses because its maturation is particularly long and susceptible to changes even after birth. The steps of this maturation overlap and form a continuum reflected in the pathology. Mental retardation occurring in children born of mothers who were subjected to atomic radiation, or ingested methylmercury from industrial waste, or who suffered the effects of lead, alcohol or tobacco constitutes a clear clinical example. Animal experiments confirm these data, adding pathogenic explanations. These experiments also explore the possible consequences that some medical techniques such as modern reproductive technology, for example embro freezing, can have on the development of the conceptus. Toxic substances or drugs can also be responsible for such abnormalities through a genetic attack on spermatogenesis. Behavioral teratogenesis also opens a larger perspective related to the optimal quality of the conceptus and the determining factors, from stress or dietary factors during apparently normal pregnancy to paternal age at the moment of conception. Finally, given that other systems than the CNS can be involved in histological or biochemical abnormalities, such as the reproductive system, we must ask what other types of functional pathology can be induced by interventions on gametes, the embryo and the fetus. Thus, behavioral teratogenesis leads to the teratogenesis of functions.

Abnormalities, Drug-Induced↗

Cell death in normo- and teratogenesis. II. Cell death in teratogenesis.

The second part of the review presents the main facts and problems with respect to the role of cell death in teratogenesis (pathogenetic cell death). After mentioning the first approaches to the domain, reference is made (in order to avoid repetition) to the synthetic article published by Menkes et al. (1970), as to a series of investigations and results up to that date. Subsequently, more recent contributions are presented, first of all a series of results obtained in the Laboratory of Embryology and Teratology, Timişcara. The genetic background and the morphology of pathogenetic cell death are briefly discussed. As a conclusion, the scheme (published in 1970) of cell death linked teratogenesis is reproduced in a somewhat modified and extended way.

Animals↗

Amelioration of teratogenesis. I. Modification of hydroxyurea-induced teratogenesis by the antioxidant propyl gallate.

Hydroxyurea (HU) is a potent teratogen which caused 100% embryotoxic effects in New Zealand white rabbits when injected sc on gestational day 12. These included a high percentage of resorptions and severe craniofacial, trunk and limb deformities of all survivors. Co-treatment of pregnant animals with the phenolic antioxidant propyl gallate (PG) resulted in amelioration of the embryotoxicity. Various amounts of PG (362-906 mg/kg) and HU (600-750 mg/kg) were eigher injected simultaneously or mixed together for periods of time up to 45 min. The extent of amelioration was dependent upon the amount of PG, although the highest dose of PG caused maternal toxicity. Simultaneous injections of HU and PG were not as efficacious as mixture of the two chemicals prior to injection. The length of time which the HU and PG mixture was allowed to stand prior to injection had no effect on the extent of amelioration. The protective action of PG resulted in significant linear reductions in both resorptions and specific malformations with increasing doses of PG. Histologic analysis of HU and HU-PG embryos disclosed that HU produced rapid cell death in the mesenchymal compartment of the embryo, particularly in the limb-buds, beginning at 2 h after treatment. Cell debris increased in amount until 8 h and remained extensive at 16 h. In contrast, HU-PG delayed the onset for cell death until 8 h. Nevertheless, at 16 h, the amount of cell debris in the limb-buds was appreciable. Thin layer chromatography (TLC) of HU-PG solution revealed no breakdown products or intermediate compounds suggesting that HU and PG do not react chemically. The presence of HU within both HU and HU-PG treated embryos was confirmed by TLC of embryonic sonicants. In contrast, TLC of embryonic extracts revealed PG only in HU-PG embryos. Light microscopy of other embryos from those same litters demonstrated extensive cell debris in the HU embryos but not in HU-PG embryos. It is suggested that the delay in onset of HU cytotoxicity is caused by the antioxidant properties of PG acting within the embryos, and that this may account for the amelioration of HU teratogenesis by PG.

Abnormalities, Drug-Induced↗

[Relations between the retinoic acid acceptor and teratogenesis of retinoids].

Retinoic acid can induce teratogenesis of the fetus of many animals including human, and its biological activities are induced by a serious of different retinoic acid accepters and their ligands. The retinoic acid acceptor RAR plays key roles in the teratogenesis, and the ligands of RAR are strong teratogens. The intensity sequence of the relative teratogenesis is ligandalpha, ligandbeta and ligandgamma. The ligands of the retinoic acid acceptor RXR cannot induce teratogenesis, but they can enhance the teratogenesis of the RAR stimulus. The retinoic acid acceptors can also affect the development of the fetus by adjusting the expression of the other genes. The relations between the gene mutation of the retinoic acid acceptor, various retinoic acid acceptors and their ligands and teratogenesis of retinoic acid are summarized in this article. In addition, the regulations of the retinoic acid acceptors to the other genes are also discussed.

Animals↗

Cytoplasmic factors do not contribute to a maternal effect on ethanol teratogenesis.

Both maternal and fetal genetic factors influence variations in response to prenatal ethanol exposure. To assess the effect of maternal genotype on the incidence of ethanol teratogenesis, a reciprocal cross study was conducted in an animal mode using the relatively susceptible C57BL/6J (B6) and the relatively resistant DBA/2J (D2) inbred mice. This mating pattern produced four embryonic genotypes: true-bred B6B6 and D2D2 litters and hybrid B6D2 and D2B6 litters. To examine the role of maternal egg cytoplasm as the source of variation that could account for a maternal effect, B6D2 and D2B6 F1 females were mated back to B6 males, which produced two additional embryonic genotypes: B6D2.B6 and D2B6.B6. Dams were intubated with either 5.8 g/kg of ethanol or an isocaloric amount of maltose-dextrin on day 9 of pregnancy. On day 18 of pregnancy, dams were sacrificed, fetuses were removed, weighed, sexed, and examined for gross morphological malformations. Every other fetus within a litter was prepared for either skeletal or soft tissue analysis. Results showed a higher rate of teratogenesis in the B6D2 group compared to the genetically similar D2B6 group, which indicates an influence of maternal genotype on susceptibility to ethanol teratogenesis. The percentage of affected male and female fetuses did not differ, which suggests that sex-linked factors are not responsible for the maternal effect. The backcross B6D2.B6 and D2B6.B6 litters did not differ significantly for any measure of teratogenesis, suggesting that differences in maternally transmitted cytoplasmic material are not the cause of the maternal effect. Factors that could account for the maternal effect are differences in the maternal uterine environment and genomic imprinting. Separating maternal from fetal-mediated mechanisms responsible for susceptibility to ethanol teratogenesis is needed for identifying mothers and infants at risk.

Abnormalities, Drug-Induced↗

Maternal genetic effects on ethanol teratogenesis and dominance of relative embryonic resistance to malformations.

Maternal genetic factors and/or fetal genetic factors contribute to variations in response to prenatal alcohol exposure. To assess the contribution of maternal genotype to ethanol teratogenesis, a reciprocal cross study was conducted in an animal model using C57BL/6J (B6) and long-sleep (LS) mice. B6 mice are more susceptible than LS mice to prenatal ethanol-induced malformations but both mouse stocks are susceptible to fetal weight deficits following in utero alcohol exposure. B6 and LS dams were reciprocally mated to B6 or LS males producing four embryonic genotype groups: the true-bred B6B6 and LSLS genotypes, and the genetically similar B6LS and LSB6 genotypes (the F1 genotype). Dams were intubated with either 5.8 g/kg ethanol (E) or an isocaloric amount of sucrose (S) on day 9 of pregnancy. Fetuses were removed on gestation day 18, weighed, and assessed for soft tissue or skeletal malformations. Results showed a greater litter weight deficit and increased total malformation rate in ethanol-exposed F1 litters carried by B6 mothers compared to ethanol-exposed F1 litters carried by LS mothers. This result would be expected only if maternal genetic factors contribute significantly towards susceptibility to ethanol teratogenesis. The influence of the LS mother was to decrease susceptibility to ethanol teratogenesis compared to the B6 mother while the influence of the B6 mother was to increase susceptibility to ethanol teratogenesis compared to the LS mother. The average malformation rate for F1 litters was significantly less than the predicted midparental value. This shows that the F1 genotype exhibited dominance towards resistance to prenatal alcohol effects.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Ethanol neurobehavioural teratogenesis and the role of L-glutamate in the fetal hippocampus.

The purpose of this article is to review the current state of knowledge of ethanol neurobehavioural teratogenesis and its postulated mechanisms. The review comprises an examination of ethanol teratogenesis in the human, including the fetal alcohol syndrome, and in experimental animals. Several current proposed mechanisms of ethanol neurobehavioural teratogenesis are critically assessed, including the role of acetaldehyde as the proximate metabolite of ethanol; fetal hypoxia; placental dysfunction; fetal prostaglandin metabolism; and action of ethanol on developing neurons in the fetal brain, including the hippocampus, one of ethanol's main target sites. The effect of ethanol on the release of L-glutamate, an excitatory amino acid neurotransmitter, in the fetal hippocampus is described, and the role of L-glutamate in ethanol teratogenesis involving the hippocampus is discussed. A novel mechanism for abnormal neuronal development in the fetal hippocampus produced by prenatal ethanol exposure is presented, and future experiments to test this hypothesis are proposed.

Animals↗

Cyclophosphamide teratogenesis: a review.

Cyclophosphamide (CP) is one of the best studied teratogens; it produces primarily central nervous system and skeletal anomalies in rats, mice, rabbits, monkeys, and humans. Furthermore, CP is one of the most extensively studied antineoplastic agents. Recent work using in vitro rodent embryo culture has demonstrated that CP must be bioactivated to be teratogenic. This finding extends earlier work showing that CP must be activated to achieve its antineoplastic and mutagenic effects. Activation of CP to its teratogenic, mutagenic, and antineoplastic form is mediated by microsomal cytochrome P-450 monooxygenases, which convert CP to 4-hydroxycyclophosphamide (4OHCP). In the absence of detoxification, 4OHCP spontaneously breaks down to phosphoramide mustard (PM) and acrolein (AC). PM is the CP metabolite believed to be responsible for the antineoplastic and mutagenic effects of CP, whereas AC is thought to cause the side effects associated with CP chemotherapy. Recent work has shown that the teratogenic effects of CP are mediated by both PM and AC. Although it is far from proven, available evidence supports the hypothesis that DNA is the primary target in terms of the teratogenic, mutagenic, and antineoplastic effects of CP. Although the nature of the DNA lesions produced by CP, which are responsible for its teratogenic, mutagenic, and antineoplastic effects, is not completely understood, cross-linking of DNA seems to play a critical role in the antineoplastic properties of CP. Preliminary information obtained from embryos exposed to CP metabolites suggests that, although DNA cross-linking might play a role in CP teratogenesis, metabolite-induced DNA strand breakage and/or induction of mutations might also play a role. Although insights into the molecular mechanisms underlying CP teratogenesis are just beginning to accumulate, the availability of in vitro embryo culture combined with the modern armamentarium of molecular biology will allow teratologists to probe further the molecular aspects of teratogenesis.

Abnormalities, Drug-Induced↗

Aspirin pretreatment potentiates hyperthermia-induced teratogenesis in the mouse.

OBJECTIVE: Our purpose was to investigate the effect of aspirin pretreatment on hyperthermia-induced teratogenesis. The rationale for the study was based on the growing evidence that prostaglandin pathway may be involved in the cellular response to the thermic injury. STUDY DESIGN: On gestation day 8.5 Swiss mice were treated with 0 or 200 mg/kg of aspirin and 1 hour later exposed to a single 10-minute thermostatic bath treatment at 38 degrees C, 41 degrees C, 42 degrees C, or 43 degrees C. On gestation day 18 uterine contents were evaluated for developmental disorders, including prenatal mortality, intrauterine growth restriction, and external, visceral, and skeletal abnormalities. RESULTS: Consistent with expectations, hyperthermia impaired morphogenesis in a dose-related manner. Although aspirin alone did not reveal embryotoxicity, its administration potentiated hyperthermia-induced teratogenesis. A statistically significant interaction (p < 0.05) was observed at 42 degrees C, where the incidence of fetuses per litter with axial skeletal malformations increased from 20.3% to 55.7%. CONCLUSION: A nonteratogenic dose of aspirin enhanced the teratogenic response to hyperthermia. This result fits the hypothesis that prostaglandins may play a protective role in hyperthermia-induced teratogenesis.

Animals↗

Effect of stiripentol dose on phenytoin-induced teratogenesis in a mouse model.

PURPOSE: Previous studies have suggested that polytherapy by design may aid in the management of human pregnancies complicated by epilepsy. However, mechanistic parallels must be drawn between the models of teratogenesis and human pregnancies, and doses of the second agent given to minimize side-effects must be justified. This study sought to determine the lowest dosage of stiripentol (STP) protective against phenytoin-induced teratogenesis in a mouse model, and to determine mechanistically if inhibition of oxidative metabolism by STP in vitro decreased production of reactive phenytoin (PHT) metabolites. METHODS: Pregnant SWV mice were assigned to control or treatment groups of STP alone, PHT alone, or PHT with ascending doses of STP coadministration. Treatments continued from Day 6 to Day 18 of gestation when fetuses were examined for developmental anomalies. [14C]PHT was incubated in mouse liver microsomes with and without NADPH and in the presence or absence of STP or piperonyl butoxide. Covalent binding of [14C] was measured. RESULTS: There were no dose-related differences in the frequency of fetal malformations per litter among groups treated with STP alone. However, STP (all doses) reduced the frequency of PHT-induced malformations. Covalent binding of [14C]PHT was NADPH-dependent and was inhibited by either piperonyl butoxide or STP. CONCLUSIONS: The beneficial effects of STP occurred at concentrations below the therapeutic range for its anticonvulsant effects. These results support the concept of polytherapy by design to reduce the risk of teratogenesis associated with PHT.

Abnormalities, Drug-Induced↗

Chromosome aberrations as a cause of subtle teratogenesis and use of the grasshopper neuroblast to test potential mutagens and teratogens.

The possible effects of low doses of environmental mutagens on the human embryo are discussed in terms of chromosome aberrations that could result in subtle teratogenesis, i.e., functional defects not detectable at birth. The action of a mutagen on the cells of an early stage human embryo has the potential of producing teratogenesis by inducing a viable chromosome aberration, e.g., a terminal deletion. Such an event would give rise to a mosaic individual. It is proposed that a functional defect of the central nervous system is the most likely result. The advantages of the neuroblast of the grasshopper embryo for detecting potential mutagens-teratogens are presented. In addition, the mitotic effects of mutagens which may also cause teratogenesis can be easily ascertained in great detail in the neuroblast, which has a short cell cycle (4 h at 38 degrees C) and which can be observed in the living condition.

Animals↗

Inhibited intercellular communication as a mechanistic link between teratogenesis and carcinogenesis.

Teratogenesis and carcinogenesis share many characteristics, leading to the speculation that they may also share pathogenic mechanisms. Direct intercellular communication mediated by membrane junctions is known to occur between a variety of cells and may play an important role in the control of cell growth and differentiation. Inhibition of junctional communication may be a mechanism common to both teratogenesis and carcinogenesis whereby cells and tissues are diverted from their normal differentiation paths. The multistage model of carcinogenesis predicts that the irreversibly initiated cell is at least partially regulated by the surrounding cells of a tissue, and that the initiated cell remains inactive until stimulated to proliferate by a tumor promotor. Tumor promoters may release the initiated cell from control of the surrounding tissue by interrupting intercellular communication, since many tumor promoters have now been shown to interfere with junctional communication in cultured mammalian cells. Furthermore, many tumorigenic cells have compromised junctional communication abilities. Similarly, it has been reasoned that the cells of an embryo must be able to communicate with each other to define tissue specificity and pattern formation, and to coordinate morphogenetic events. Many studies have chronicled alterations in junctional communication that occur coincident with major developmental events and some studies suggest that junctional communication may be modified at boundaries of morphogenetic fields. A recent in vivo study has provided evidence that inhibition of junctional communication may interfere with embryonic development, and several teratogens are known to interrupt junctional communication in mammalian cells in culture. These observations suggest that inhibition of junctional intercellular communication may be a shared mechanism of carcinogenesis and teratogenesis.

Animals↗

Evaluation of the developmental toxicity of thalidomide using frog embryo teratogenesis assay-xenopus (FETAX): biotransformation and detoxification.

The developmental toxicity of thalidomide was evaluated using FETAX (Frog Embryo Teratogenesis Assay - Xenopus). Young X. Laevis embryos were exposed to this compound in each of two concentration-response experiments with and without differently induced exogenous metabolic activation systems (MASs) and/or inhibited MASs. Young male Sprague-Dawley rats were treated with either isoniazid or Aroclor 1254 to induce cytochrome P-450. Several of the rats were subsequently treated with diethyl maleate (DM) to deplete glutathione reserves. Specific aliquots of rat liver microsomes were treated with 3-amino-1,2,4-triazole (ATZ) or alpha-napthoflavone (alpha-N) to selectively inhibit P-450 activity. Bioactivation was indicated by increased developmental toxicity observed in MAS tests. Results obtained indicated that thalidomide was predominantly activated by P-450 isozyne CYP2E1, although weak cross-specificity between CYP1A1/A2 may have existed. Detoxification pathways for thalidomide were investigated by treatment of the MAS with cyclohexene oxide (CHO) and DM to inhibit the epoxide hydrolase and glutathione conjugation pathways, respectively. Results indicated that epoxide hydrolase was primarily responsible for the detoxification of bioactivated thalidomide. Teratogenesis Carcinog. Mutagen. 20:35-47, 2000.

Amitrole↗

The role of inhibited cell-cell communication in teratogenesis.

A mechanistic link between teratogenesis and carcinogenesis has been suggested by a wide variety of scientific observations. This report attempts to provide a theoretical explanation for one of the several possible mechanisms which might be shared during carcinogenesis and teratogenesis. The initiation and promotion concept of carcinogenesis was briefly reviewed and the role of intercellular communication during the complex tumor promotion phase was discussed. Inhibition of intercellular communication by a wide variety of physical, chemical and biological factors was speculated to disrupt the regulation of proliferation and differentiation in stem cells. Chemicals, which interfered with intercellular communication during early organogenesis, have the potential of being teratogens, while if they are present in the developed, initiated organisms have the potential of being tumor promoters. Evidence was presented showing that known tumor promoters which inhibited intercellular communication also had been shown to be teratogens. It was concluded that in vitro assays, designed to measure intercellular communication, although having known limitations, might be used as an in vitro means to screen for potential teratogens.

Abnormalities, Drug-Induced↗

A selection of candidate compounds for in vitro teratogenesis test validation.

The Consensus Workshop on In Vitro Teratogenesis Testing recommended that test validation be facilitated by a listing of agents with defined teratogenicity; subsequently, a panel was convened to review and select such agents. This communication established a list of 47 compounds or conditions which demonstrate a wide range of teratogenicity in vivo. The agents were chosen primarily on the strength of the literature base denoting their in vivo effects. The tables note a number of general biological and toxicological characteristics for each agent, and the details of representative in vivo teratology studies are summarized and referenced. This list is intended to serve as a base for in vitro teratogenesis test validation and should prove useful in developing and identifying those systems which will contribute to a more effective testing program.

Abnormalities, Drug-Induced↗

Development of a reconstituted water medium and preliminary validation of the frog embryo teratogenesis assay--Xenopus (FETAX).

A reconstituted water medium was developed for use in the Frog Embryo Teratogenesis Assay--Xenopus (FETAX). FETAX solution was then tested on three compounds with known mammalian teratogenicity (ethanol, caffeine, and 5-fluorouracil) as was a non-teratogen (saccharin). The results obtained were then compared with results from tests on these compounds in two other media that had previously been used in the assay. Saccharin was not teratogenic. Ethanol and caffeine were weak and moderate teratogens, respectively. 5-fluorouracil was a strong teratogen. The results compare favorably with those obtained in mammalian studies. The amount of growth inhibition in embryos in the 96 h tests was positively correlated with the degree of teratogenicity of the compound. Final validation of FETAX will allow it to be used to screen and rank compounds for further testing and as a tool for studying the basic mechanisms of teratogenesis.

Animals↗