Genesis of alcohol-induced craniofacial dysmorphism.

The initial diagnosis of fetal alcohol syndrome (FAS) in the United States was made because of the facial features common to the first cohort of patients. This article reviews the development of an FAS mouse model whose craniofacial features are remarkably similar to those of affected humans. The model is based on short-term maternal treatment with a high dosage of ethanol at stages of pregnancy that are equivalent to Weeks 3 and 4 of human gestation. At these early stages of development, alcohol's insult to the developing face is concurrent with that to the brain, eyes, and inner ear. That facial and central nervous system defects consistent with FAS can be induced by more "realistic" alcohol dosages as illustrated with data from an oral alcohol intake mouse model in which maternal blood alcohol levels do not exceed 200 mg/dl. The ethanol-induced pathogenesis involves apoptosis that occurs within 12 hrs of alcohol exposure in selected cell populations of Day 7, 8, and 9 mouse embryos. Experimental evidence from other species also shows that apoptosis underlies ethanol-induced malformations. With knowledge of sensitive and resistant cell populations at specific developmental stages, studies designed to identify the basis for these differing cellular responses and, therefore, to determine the primary mechanisms of ethanol's teratogenesis are possible. For example, microarray comparisons of sensitive and resistant embryonic cell populations have been made, as have in situ studies of gene expression patterns in the populations of interest. Studies that illustrate agents that are effective in diminishing or exacerbating ethanol's teratogenesis have also been helpful in determining mechanisms. Among these agents are antioxidants, sonic hedgehog protein, retinoids, and the peptides SAL and NAP.

Protection from ethanol-induced limb malformations by the superoxide dismutase/catalase mimetic, EUK-134.

Based on previous in vitro studies that have illustrated prevention of ethanol-induced cell death by antioxidants, using an in vivo model, we have tested the anti-teratogenic potential of a potent synthetic superoxide dismutase plus catalase mimetic, EUK-134. The developing limb of C57BL/6J mice, which is sensitive to ethanol-induced reduction defects, served as the model system. On their ninth day of pregnancy, C57BL/6J mice were administered ethanol (two intraperitoneal doses of 2.9 g/kg given 4 h apart) alone or in combination with EUK-134 (two doses of 10 mg/kg). Pregnant control mice were similarly treated with either vehicle or EUK-134, alone. Within 15 h of the initial ethanol exposure, excessive apoptotic cell death was observed in the apical ectodermal ridge (AER) of the newly forming forelimb buds. Forelimb defects, including postaxial ectrodactyly, metacarpal, and ulnar deficiencies, occurred in 67.3% of the ethanol-exposed fetuses that were examined at 18 days of gestation. The right forelimbs were preferentially affected. No limb malformations were observed in control fetuses. Cell death in the AER of embryos concurrently exposed to ethanol and EUK-134 was notably reduced compared with that in embryos from ethanol-treated dams. Additionally, the antioxidant treatment reduced the incidence of forelimb malformations to 35.9%. This work illustrates that antioxidants can significantly improve the adverse developmental outcome that results from ethanol exposure in utero, diminishing the incidence and severity of major malformations that result from exposure to this important human teratogen.

Fear conditioning-induced alterations of phospholipase C-beta1a protein level and enzyme activity in rat hippocampal formation and medial frontal cortex.

We investigated the effects of one-trial fear conditioning on phospholipase C-beta1a catalytic activity and protein level in hippocampal formation and medial frontal cortex of untreated control rats and rats prenatally exposed to ethanol. One hour following fear conditioning of untreated control rats, phospholipase C-beta1a protein level was increased in the hippocampal cytosolic fraction and decreased in the hippocampal membrane and cortical cytosolic and cortical membrane fractions. Twenty-four hours after fear conditioning, phospholipase C-beta1a protein level was reduced in the hippocampal cytosolic fraction and elevated in the cortical nuclear fraction; in addition, 24 h after conditioning, phospholipase C-beta1a activity in the cortical cytosolic fraction was increased. Rats that were exposed prenatally to ethanol displayed attenuated contextual fear conditioning, whereas conditioning to the acoustic-conditioned stimulus was not different from controls. In behavioral control (unconditioned) rats, fetal ethanol exposure was associated with reduced phospholipase C-beta1a enzyme activity in the hippocampal nuclear, cortical cytosolic, and cortical membrane fractions and increased phospholipase C-beta1a protein level in the hippocampal membrane and cortical cytosolic fractions. In certain cases, prenatal ethanol exposure modified the relationship between fear conditioning and changes in phospholipase C-beta1a protein level and/or activity. The majority of these effects occurred 1 h, rather than 24 h, after fear conditioning. Multivariate analysis of variance revealed interactions between fear conditioning, subcellular fraction, and prenatal ethanol exposure for measures of phospholipase C-beta1a protein level in hippocampal formation and phospholipase C-beta1a enzyme activity in medial frontal cortex. In the majority of cases, fear conditioning-induced changes in hippocampal phospholipase C-beta1a protein level were augmented in rats prenatally exposed to ethanol. In contrast, fear conditioning-induced changes in cortical phospholipase C-beta1a activity were, often, in opposite directions in prenatal ethanol-exposed compared to diet control rats. We speculate that alterations in subcellular phospholipase C-beta1a catalytic activity and protein level contribute to contextual fear conditioning and that learning deficits observed in rats exposed prenatally to ethanol result, in part, from dysfunctions in phospholipase C-beta1a signal transduction.

Fetal alcohol syndrome: a Japanese perspective.

To estimate and prevent the effects of prenatal alcohol on the central nervous system (CNS), brain dysfunction in fetal alcohol syndrome (FAS) and fetal alcohol effects (FAE) was compared by both epidemiological and experimental studies. The FAS infants exhibited a more severe degree of CNS involvement than the FAE infants. The CNS involvement features were developmental delay and intellectual impairment in both FAS and FAE. The increased risk of low birth weight and CNS involvement were much more significant in women who were heavy drinkers or alcoholics and smoked. The beneficial effect of supplementary zinc on the fetal cerebrum of FAS or FAE rats was limited, never reaching the unexposed control level. One of the most vulnerable structures in the rat fetus exposed to ethanol in utero was the synaptic formation in the hippocampus. The consistent dysmorphogenesis of synapses during early brain development may be associated with the functional impairment of the CNS in FAS and FAE.

[Maternal environment and developmental brain damages].

To estimate and to prevent the developmental brain damage caused by maternal environmental agents, the results were compared between epidemiological and experimental studies. I. Ethanol. The features of CNS involvement in human FAS (fetal alcohol syndrome) and FAE (fetal alcohol effects) were developmental delay and intellectual impairment. In rat FAS or FAE, a beneficial effect of supplementary zinc on the fetal cerebrum was observed, but was limited. The consistent dysformation of synapses during early brain development may be associated with the functional impairment of CNS in FAS and FAE II. Caffeine. A reduction in the fetal cerebral weight was observed with caffeine ingestion during pregnancy at levels of 1.5-3.0 micrograms caffeine/ml or g wet wt. in dams and fetuses. Maternal caffeine disturbs the neonatal cerebrum and produces behavioral abnormalities in developing rats. III. X-irradiation. Our study provides evidence of the protection by vitamin E of neuronal development in X-irradiated rat fetuses, through its antioxidant properties, against attacks by free radicals and/or lipid peroxide. IV. Low copper level in brain. A teratogenic effect of triethylene tetramine dihydrochloride, a chelating drug for copper, on the mouse brain was noted, both grossly and microscopically, with a dose of higher than 6,000 micrograms/l, which is twenty times as high as the clinical treatment dose. V. Tobacco. Thirty-three percent of human FTS (fetal tobacco syndrome) cases had CNS involvement, which was characterized by developmental delay. Fifty percent or more of FTS children of women who smoked more than 20 cigarettes per day had CNS involvement. In conclusion, the concomitant effects of maternal environmental agents on human brain development should be studied.

[Disorders of brain development in embryos obtained from mothers who abused alcohol]. / Narusheniia razvitiia golovnogo mozga émbrionov, poluchennykh ot zloupotrebliavshikh alkogolem materei.

Nineteen 5 to 12-weeks-old embryos from women abusing alcohol during pregnancy (alcoholism, habitual drinking) were studied. In 15 (79%) the brain development disorders were detected. Most frequently concerned were the brain hemispheres. Defective development signs were also noted in the midbrain, choroidal plexus and diencephalon. Microdysplasias were found in different brain regions. In one case embryonal signs of the capsula interna, nucleus caudatus, putamen, epiphysis cerebri, habenular nuclei, chiasma, optical tracts were absent. Six groups presenting most typical deviations were identified. The degree of embryonal brain disorders correlated with its mother's alcohol abuse severity.