High Doses Of Ethanol Research Articles

Acetaldehyde accumulation in rat mammary tissue after an acute treatment with alcohol

Previous studies reported the presence in rat mammary tissue of a cytosolic xanthine oxidoreductase pathway for the metabolism of alcohol to acetaldehyde and hydroxyl radicals and to the microsomal biotransformation of ethanol to acetaldehyde. It was also reported that after chronic ethanol drinking stressful oxidative conditions can be observed. The present work reports that even after single doses of ethanol, given at three different levels (6.3 g kg(-1); 3.8 g kg(-1) or 0.6 g kg(-1) p.o.), acetaldehyde accumulates for prolonged periods of time in the mammary tissue to reach concentrations higher than in blood (e.g. 5.1+/-1.2 nmol g(-1) versus 0.2+/-0.1 nmol ml(-1), for 6.3 g kg(-1) dose, 6 h after intoxication). The presence in rat mammary tissue of low activities of additional enzymes able to generate acetaldehyde was established (alcohol dehydrogenase: 0.97+/-0.84 mU mg(-1) protein; CYP2E1: 1.30+/-0.12 x 10(-2) pmol 4-nitrocatechol min(-1) mg(-1) protein) and a low activity of aldehyde dehydrogenase was observed in the cytosolic, mitochondrial and microsomal fractions (0.02+/-0.04; 0.35+/-0.09 and 0.72+/-0.19 mU mg(-1) protein, respectively). After a single high dose of ethanol, an increased susceptibility to oxidative stress was observed, as evidenced by changes in the shape of t-butylhydroperoxide induced emission of chemiluminescence in mammary tissue (6.3 g kg(-1) dose; at 3 and 6 h). In summary, the results show that even after single doses of ethanol, acetaldehyde, either formed in situ or arriving via blood, tends to accumulate in mammary tissue and that this condition might decrease cell defenses against injury.

Effects of alcohol on psychomotor performance and perceived impairment in heavy binge social drinkers

Alcohol intoxication often results in negative consequences; however, specific behavioral and subjective effects vary as a function of individual differences. The present study utilized an alcohol challenge paradigm to examine whether heavy binge social drinkers (HD; n = 77), compared to light social drinkers (LD; n = 55), exhibit: (1) greater tolerance in psychomotor task performance under the influence of alcohol, and (2) differential perceptions of the impairing effects of alcohol. The study included three test sessions in which participants consumed either a low (0.4 g/kg) or a high (0.8 g/kg) dose of ethanol or a placebo beverage administered in random order and counterbalanced within group. Participants completed the Digit-Symbol Substitution Task (DSST) and the Grooved Pegboard at pre-drink baseline and at multiple time points after beverage consumption. They also completed a scale of perceived impairment at several intervals after beverage consumption. Ethanol impaired performance at the high dose, but not at the low dose ( ps < .001). The groups exhibited similar alcohol-induced impairment. However, HD reported lower self-perceived impairment compared to LD, particularly during the early portion of the blood alcohol curve when actual impairment was most pronounced ( p < .001). Thus, this study extends prior research in that habitual binge social drinking does not appear to be associated with tolerance to alcohol's impairing effects on select psychomotor skills. Further, results may have implications for alcohol-related harm as binge social drinkers regularly consume intoxicating doses of alcohol but may not be aware of the physical and cognitive impairments produced by alcohol.

Acute Ethanol Administration Rapidly Increases Phosphorylation of Conventional Protein Kinase C in Specific Mammalian Brain Regions in Vivo

Protein kinase C (PKC) is a family of isoenzymes that regulate a variety of functions in the central nervous system including neurotransmitter release, ion channel activity, and cell differentiation. Growing evidence suggests that specific isoforms of PKC influence a variety of behavioral, biochemical, and physiological effects of ethanol in mammals. The purpose of this study was to determine whether acute ethanol exposure alters phosphorylation of conventional PKC isoforms at a threonine 674 (p-cPKC) site in the hydrophobic domain of the kinase, which is required for its catalytic activity. Male rats were administered a dose range of ethanol (0, 0.5, 1, or 2 g/kg, intragastric) and brain tissue was removed 10 minutes later for evaluation of changes in p-cPKC expression using immunohistochemistry and Western blot methods. Immunohistochemical data show that the highest dose of ethanol (2 g/kg) rapidly increases p-cPKC immunoreactivity specifically in the nucleus accumbens (core and shell), lateral septum, and hippocampus (CA3 and dentate gyrus). Western blot analysis further showed that ethanol (2 g/kg) increased p-cPKC expression in the P2 membrane fraction of tissue from the nucleus accumbens and hippocampus. Although p-cPKC was expressed in numerous other brain regions, including the caudate nucleus, amygdala, and cortex, no changes were observed in response to acute ethanol. Total PKCgamma immunoreactivity was surveyed throughout the brain and showed no change following acute ethanol injection. These results suggest that ethanol rapidly promotes phosphorylation of cPKC in limbic brain regions, which may underlie effects of acute ethanol on the nervous system and behavior.

Metabolic interaction between ethanol, high-dose alprazolam and its two main metabolites using human liver microsomes in vitro

Alprazolam is widely used as a short-acting antidepressant and anxiolytic agent and its effect appears at very low doses while ethanol is used as a social drug worldwide. Sometimes, toxic interactions occur following combined administration of these two drugs. In this study we have investigated the interaction between ethanol and high-dose alprazolam using human liver microsomes in vitro. The interaction effects between ethanol and alprazolam were examined by a mixed-function oxidation reaction using a human liver microsomal preparation. Alprazolam and its two main metabolites (α-hydroxyalprazolam: α-OH alprazolam, 4-hydroxyalprazolam: 4-OH alprazolam) were measured by HPLC/UV. The production of 4-OH alprazolam, one main metabolite of alprazolam, was weakly inhibited by higher dose of ethanol, but not α-OH alprazolam. These results using a human liver microsomal preparation show that the production of 4-OH alprazolam is weakly inhibited by ethanol but not α-OH alprazolam. Toxic levels may be reached by simultaneous administration of ethanol and high-dose alprazolam.

Chronic swim stress alters sensitivity to acute behavioral effects of ethanol in mice

Epidemiological data support a strong link between stress, stress-related disorders and risk for alcoholism. However, precisely how stress might impact sensitivity to the intoxicating effects of ethanol or the willingness to voluntary consume ethanol remains unclear. The present study assessed the effects of daily exposure to forced swim stress on subsequent sensitivity to the sedative/hypnotic, hypothermic, ataxic (measured using accelerating rotarod), and anxiolytic-like (measured using elevated plus-maze) effects of ethanol, and ethanol consumption and preference in a two-bottle choice paradigm, in male C57BL/6J mice. Stress effects on the sedative/hypnotic effects of the barbiturate pentobarbital were also tested. Results showed that chronic (fourteen days) but not acute (one or three days) swim stress significantly potentiated the sedative/hypnotic and hypothermic effects of 4 g/kg, but not 3 g/kg, ethanol. The sedative/hypnotic effects of pentobarbital were attenuated by chronic swim stress. Irrespective of chronicity, swim stress did not alter the ataxic or anxiolytic-like effects of ethanol, or alter ethanol self-administration either during or after stress. These data provide further evidence that stress alters the intoxicating effects of high doses of ethanol in a behaviorally selective manner.

Ethanol Inhibits the Sensory Responses of Cerebellar Granule Cells in Anesthetized Cats

Granule cells occupy a strategic position in the transmission of afferent information to the cerebellar cortex. They are also the most abundant type of neurons in the cerebellum. The functions of the cerebellum are thought to be sensitive to acute alcohol intoxication. The effects of acute alcohol intoxication on the in vivo physiology of cerebellar granule cells are, however, not completely known. We studied chloralose-anesthetized cats at ethanol doses relevant to human drinking (0.3-1.2 g/kg). We recorded the electrophysiological responses of granule cell clusters to auditory and visual stimulation, and simultaneously monitored the concentration of ethanol in the cerebrospinal fluid (CSF). At an intravenous ethanol dose of 0.3 g/kg, CSF ethanol concentration peaked in 10 minutes at 17 mM, equivalent to a blood alcohol concentration (BAC) of about 0.08 g/dL. Ethanol quickly and almost completely abolished both auditory and visual responses from granule cells. Complete or near-complete inhibition lasted 15 to 20 minutes; approximately 50% recovery required an additional 15 minutes, and a full recovery yet another 15 minutes. A higher ethanol dose at 1.2 g/kg resulted in a more severe inhibition and required longer time for recovery. The relationship between ethanol dose, CSF ethanol concentration, and granule cell responses was dynamic and nonlinear, critically depending upon the elapsed time. Cerebellar granule cell sensory responses are highly sensitive to ethanol inhibition. A rapid development of acute tolerance appears to be a major factor contributing to the dynamic and nonlinear relationship among ethanol dosage, CSF ethanol concentration, and granule cell responses. It is likely that a generalized de-afferentation of the cerebellum from its mossy fiber afferents, followed by the subsequent development of acute tolerance may play major roles by which alcohol intoxication affects cerebellar functions.

Transdermal absorption of the herbicide 2,4-dichlorophenoxyacetic acid is enhanced by both ethanol consumption and sunscreen application

Xenobiotics absorption is a health concern and skin is a major exposure site for many of these chemicals. Both alcohol consumption and topical sunscreen application act as transdermal penetration enhancers for model xenobiotics. The effect of combining these two treatments on transdermal absorption of the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) was therefore examined. Skin from rats ingesting low (1.5 g/kg) medium (4.3 g/kg) or high (6 g/kg) ethanol doses or saline control was treated with a commercially available sunscreen containing titanium dioxide and octyl methoxycinnimate and transdermal absorption of 2,4-D was monitored. Ethanol increased penetration by a factor of 1.9, 2.0 and 2.5 for animals treated with 1.5, 4.3 and 6 g/kg respectively, demonstrating an ethanol-induced dose response. Sunscreen application to skin from ethanol gavaged rats caused 2,4-D absorption above that induced by ethanol alone by an additional factor of 1.3, 2.1 and 2.9 for 1.5, 4.3 and 6 g/kg respectively. Comparing 2,4-D transdermal absorption after exposure to both ethanol and sunscreen with a theoretical value (sum of penetration after ethanol or sunscreen treatment) demonstrates that these two treatments enhance additively at the higher doses tested. Results of this study emphasize the importance of limiting excessive alcohol consumption in individuals with potential herbicide exposure rather than discouraging the use of sunscreens, since the consequences of UV-induced skin cancer are far more series than the risks that would be associated with observed increases in chemical exposure.

Effects of acute and chronic ethanol exposure on the behavior of adult zebrafish ( Danio rerio)

The zebrafish has been a popular subject of embryology and genetic research for the past three decades. Recently, however, the interest in its neurobiology and behavior has also increased. Nevertheless, compared to other model organisms, e.g., rodents, zebrafish behavior is understudied and very few behavioral paradigms exist for mutation or drug screening purposes. Alcoholism is one of the biggest and costliest diseases whose mechanisms are not well understood. Model organisms such as the zebrafish may be utilized in this line of research. Previously, we investigated the effects of acute ethanol exposure on adult zebrafish using four behavioral paradigms and employing manual quantification methods. Here, we study the effects of chronic ethanol exposure and analyze how it modifies the effects of acute ethanol treatment. We employ a videotracking-based automated quantification method in a predator model paradigm and show that this method is capable of detecting an avoidance reaction that is ameliorated by higher doses of ethanol, a potential anxiolytic effect. Importantly, we also demonstrate that chronic, two week long, exposure to ethanol results in significant adaptation to this substance in adult zebrafish. Overall, our results suggest that zebrafish will be an appropriate subject for high throughput screening applications aimed at the analysis of the mechanisms and pharmacology of acute and chronic ethanol induced changes in the vertebrate brain.

The influence of ethanol intake and its withdrawal on the anticonvulsant effect of α-tocopherol in the penicillin-induced epileptiform activity in rats

Previous studies proposed the existence of a relationship between epilepsy and ethanol. Ethanol may have either proconvulsive or anticonvulsive effects on epileptic activity in different experimental epilepsy models. The influence of high dose ethanol intake and its withdrawal on the anticonvulsant effect of α-tocopherol was examined after intracortical injection of penicillin (500 units) to induce epileptiform activity. Thirty minutes after penicillin injection, the most effective dose of α-tocopherol (500 mg/kg) was administrated intramuscularly (i.m.). Ethanol-treated rats received a daily dose of 9.0 g/kg of 30% ethanol solution via an oesophageal probe for 15 days. All rats in the withdrawal group were anesthetized for induction of penicillin-induced epileptiform activity 28 h after the last ethanol administration. The epileptiform activity was verified by electrocorticographic (ECoG) recordings. Ethanol, in a dose of 9 g/kg, significantly decreased the mean frequency of penicillin-induced epileptiform ECoG activity without changing the amplitude. The mean frequency of ECoG activity was decreased in the 60 and 70 min period from penicillin injection in the ethanol-treated + α-tocopherol and ethanol withdrawal + α-tocopherol groups compared with the penicillin-injected (500 units, i.c.) group, respectively. α-Tocopherol was more effective in decreasing the mean frequency of epileptiform activity in the ethanol + α-tocopherol group than in other α-tocopherol administrated groups. Ethanol withdrawal caused an increase in frequency of epileptiform activity in the withdrawal + α-tocopherol group compared with other α-tocopherol administrated groups. α-Tocopherol did not affect the amplitude of epileptiform activity in any group. Possible mechanisms of ethanol influence on the neuroprotective actions of α-tocopherol are still a crucial issue associated with epilepsy.

Differential Effects Between Maotai and Ethanol on Hepatic Gene Expression in Mice: Possible Role of Metallothionein and Heme Oxygenase-1 Induction by Maotai

Alcohol is a risk factor for liver fibrosis and hepatocellular carcinoma. On the other hand, light alcoholic beverage consumption is believed to be beneficial because of the effects of both alcohol and nonalcoholic components of the beverage. Maotai is a commonly consumed beverage in China containing 53% alcohol. Epidemiological and experimental studies show that Maotai is less toxic to the liver than ethanol alone. To examine the differential effects of Maotai and ethanol, a low dose of Maotai or an equal amount of ethanol (53%, v/v in water, 5 ml/kg) were given to male mice daily for 1 week, and hepatic RNA was extracted for microarray analysis. Approximately 10% of genes on the liver-selective custom array (588 genes) were altered following Maotai or ethanol administration, but Maotai treated livers had fewer alterations compared with ethanol alone. Real-time reverse transcription-polymerase chain reaction confirmed and extended microarray results on selected genes. An induction of metallothionein and heme oxygenase-1 occurred with Maotai, which could not be explained by alcohol consumption alone, whereas the attenuation of ethanol responsive genes such as quinone dehydrogenase, DNA-ligase 1, IGFBP1, and IL-1beta suggests less liver injury occurred with Maotai. The expression of genes related to liver fibrosis, such as cytokeratin-18, was slightly increased by the high dose of ethanol, but was unchanged in the Maotai group. In summary, gene expression analysis indicates that Maotai induces a different response than ethanol alone. The dramatic induction of metallothionein and heme oxygenase-1 with Maotai could be important adaptive responses to reduce alcoholic liver injury.

Ethanol exposure differentially alters pro-enkephalin mRNA expression in regions of the mesocorticolimbic system

Opioid peptides have been suggested to play a major role in ethanol reinforcement mechanisms and alcohol drinking behaviour. However, in non-selected strains of rodents, it is not known whether opioid biosynthesis is a critical event in these processes. The aim of this work was to study the effects of a high dose of ethanol (2.5 g/kg body weight) on pro-enkephalin (pro-enk) mRNA expression in brain regions of the mesocorticolimbic system for up to 24 h after drug administration. Male Wistar rats were administered with ethanol (2.5 g/kg body weight) or distilled water and were killed 30 min, 1, 2, 4, 8 or 24 h after treatment. Coronal brain sections (20 mu) were obtained and pro-enk mRNA expression was studied by in situ hybridization and densitometry. Acute ethanol administration induced a transient decrease and increase in pro-enk mRNA expression in the ventral tegmental area (33.2%) and prefrontal cortex (26.5%) 2 and 4 h after treatment, respectively. In contrast, ethanol induced prolonged increases in pro-enk mRNA expression in the core and shell regions of the nucleus accumbens, with different kinetics. Maximal effects were observed 2 h after ethanol exposure (core, 70.0%; shell, 60.0%). Our results indicate that enkephalin expression in regions of the rat mesocorticolimbic system is differentially altered by acute ethanol treatment and suggest that enkephalins may play a key role in ethanol reinforcement mechanisms.

Gender specific effects of ethanol in mice, lacking CCK 2 receptors

Neuropeptide cholecystokinin (CCK) has been reported to suppress ethanol intake, but there is contradictory evidence about the role of CCK 2 receptors. In the present study anxiolytic, hypolocomotor and sedative effects of acute ethanol administration, but also voluntary ethanol consumption were studied in male and female mice, lacking CCK 2 receptors (−/−). Ethanol (1.0 and 2.0 g/kg) induced a significant reduction of anxiety-related behaviours in the elevated plus-maze, but this effect was statistically significant only in female homozygous mice (−/−). In male mice, lacking CCK 2 receptors (−/−), but not in their wild-type littermates (+/+), the suppression of vertical locomotor activity was caused by ethanol at a dose 0.5 g/kg. The highest dose of ethanol (2.0 g/kg) produced statistically significant reduction of horizontal locomotor activity only in female wild-type (+/+) mice, but this effect was related to increased basal activity when compared to female mutant (−/−) mice. Duration of the loss of righting reflex was not significantly affected by genotype or gender, but blood ethanol levels at regain of righting reflex were significantly lower in female homozygous mice (−/−) compared to their wild-type (+/+) littermates, indicating increased sensitivity to the sedative effect of ethanol. Ethanol intake, but not preference, at concentration 10% was significantly increased in female mice, lacking CCK 2 receptors (−/−). The present study revealed an altered response to the acute effects of ethanol in CCK 2 receptor deficient mice (−/−). These changes are gender-specific and could be attributed to the altered activity of dopaminergic system in male mice and increased activity of GABA-ergic system in female mice as established in our previous studies.

Ethanol sensitization in a neurodevelopmental lesion model of Schizophrenia in rats

Substance use disorder comorbidity in schizophrenia may reflect dysfunctional cortical-striatal-limbic circuitry commonly involved in the addiction process and the pathogenesis of schizophrenia. Rats with neonatal ventral hippocampal lesions (NVHL) demonstrate post-adolescent onset of schizophrenia-like symptoms and increased addiction vulnerability in paradigms using cocaine in adulthood. Here, we investigated response profiles of young adult NVHL vs. SHAM rats to ethanol, an addictive drug with many psychopharmacological effects divergent from those of cocaine, in a locomotor sensitization paradigm. Over 15 days of daily injections of saline, low (0.15 g/kg) or high (1.0 g/kg) doses of ethanol, NVHL rats showed stimulatory effects at the low dose compared to saline and high-dose conditions, while SHAM rats showed expected patterns of dose-dependent suppression of locomotor activity. In a challenge session 2 weeks later in which a moderate dose (0.25 g/kg) of ethanol was given to all subjects, NVHL rats with history of prior ethanol exposure showed greater locomotor activity consistent with installment of alcohol-induced sensitization not present in SHAMs. These findings provide further evidence of enhanced short- and long-term responsivity to abused drugs in a neurodevelopmental model of schizophrenia, and may reflect potentiation of common mechanisms of addiction shared between pharmacologically diverse addictive drugs.

Infantile Sensitivity to Ethanol's Motivational Effects: Ethanol Reinforcement During the Third Postnatal Week

Although tests specific to newborn rats have frequently verified their susceptibility to the reinforcing properties of ethanol, demonstration of comparable reinforcing effects in older infants has been elusive. Using a second-order conditioning procedure, the present study assessed in preweanling rats whether pairing with early postabsorptive effects of ethanol would render intraorally delivered gustatory stimuli capable of positive reinforcement for association with a salient texture. Direct reinforcing effects of ethanol were also evaluated through intake tests of gustatory stimuli previously paired with the drug. Blood ethanol levels (BELs) were determined for each of the ethanol doses used. Pups (14 days old) were stimulated with intraoral infusion of sucrose (10% v/v), water, or quinine (0.0045% w/v) 5 minutes after being intragastrically (i.g.) administered 0.00, 0.25, 0.50, or 2.00 g/kg ethanol (Experiments 1 and 2). These stimuli were then briefly presented while pups experienced a rough texture (sandpaper). Rats were subsequently evaluated in a 2-way texture location test (sandpaper vs smooth surface). In Experiment 3, sucrose, water, or quinine was paired with early postabsorptive effects of ethanol (0.00, 0.50, or 2.0 g/kg). Consumption of these stimuli was later assessed. Motor activity patterns during the intake test were also evaluated. In Experiment 4, BELs corresponding to 0.25, 0.50, or 2.0 g/kg ethanol were determined 5 and 20 minutes after i.g. administration (time periods were in accord with the onset and offset of intraoral stimulation used in the previous experiments). Intraoral infusion of sucrose, water, or quinine, while under a state of sobriety and paired with sandpaper, resulted in roughly 50% preference for this texture. Sandpaper preferences were significantly elevated in pups that had experienced sucrose or water in a nonsober state-while under the effects of ethanol (Experiments 1 and 2). This indicated reinforcing effects of the ethanol intoxication. Pairing ethanol intoxication directly with consumption of sucrose, water, or quinine did not affect their later consumption. Yet, there were clear indications that this pairing resulted in conditioned behavioral activity patterns. Blood ethanol levels corresponding to the ethanol doses used here ranged between 10 and 150 mg%. Infants appear sensitive to pharmacological reinforcing properties of low and relatively high ethanol doses. This sensitivity was revealed indirectly, by pairing gustatory stimuli with ethanol intoxication and then allowing these stimuli to act as second-order reinforcement for a quite different (tactile) stimulus. Behavioral activation elicited by the gustatory stimuli previously paired with a state of intoxication seems to compete with the expression of ethanol's motivational properties as assessed through intake tests.

Interactions between prenatal ethanol exposure and postnatal learning about ethanol in rat pups

Exposure to ethanol during the last days of gestation (1 or 2/kg) has been shown to induce greater ethanol intake as well as enhanced ethanol palatability in infant rats compared to pups without previous experience with the drug. This higher acceptance of ethanol seems to result from the prenatal association between the chemosensory aspects of ethanol and its reinforcing properties; the latter mediated at least in part by the opioid system. A series of experiments analyzed how this prenatal experience with ethanol affected postnatal conditioning of the flavor of ethanol. In the first experiment, pups exposed prenatally to ethanol (2 g/kg during gestational days 17–20) were tested for acquisition and extinction of a conditioned aversion to the taste of ethanol (conditioned stimulus), employing lithium chloride as the aversive unconditioned stimulus. In the next two experiments, pups with the same prenatal exposure to ethanol were tested for the extinction of an aversion, employing three (Experiment 2) or one (Experiment 3) conditioning trials, using a high dose of ethanol (3 g/kg) as the aversive agent. The first experiment showed that prenatal ethanol exposure delayed the postnatal acquisition of a conditioned aversion to the taste of ethanol. In the next two experiments, the same effect was observed during the extinction trials. These results provide additional support to the hypothesis of an appetitive associative memory established prenatally as a consequence of fetal ethanol exposure.

Secretion rate and amylase concentration of whole saliva after consumption of beer

Saliva is of paramount importance for the maintenance of oral health (1). Only a few studies have investigated the acute effects of alcohol consumption on salivary function. In rats, acute administration of a high dose of ethanol caused a significant reduction in stimulated parotid secretion (2) and reduced the protein synthesis in all major salivary glands (3). Ingestion of a high dose of ethanol by humans also caused a decrease in parotid salivary flow rate (4) and stimulated whole saliva flow rate (5), with a concomitant reduction in salivary amylase activity and output (5). In this study, we investigated the possible effects of a relatively small volume of beer on salivary function. Ten healthy volunteers (seven men and three women; mean age 32.1 ± 17.9 years) participated in this study. They were instructed to abstain from alcohol at least 12 h prior to the experiment and to abstain from smoking, eating, drinking caffeine containing beverages and toothbrushing at least 1 h prior to the experiment. All experiments took place between 09:30 and 12:30 hours. In a crossover design, the volunteers consumed either 300 ml top-fermented beer (Palm Breweries, Steenhuffel, Belgium, 5.2% alcohol) within 5 min. As a control, 300 ml non-alcoholic beer was used (Amstel Malt, Zoeterwoude, The Netherlands, <0.1% alcohol). The time interval between both experimental conditions was 1 week. Immediately before the consumption of beer and 15, 30, 45, 60, 90 and 120 min later, mechanically stimulated whole saliva was collected in preweighed tubes for 5 min by parafilm-chewing (6). The salivary flow rates were determined gravimetrically. SHORT COMMUNICATION

Effect of a single dose of ethanol on developing peripheral nerve of chick embryos

Fetal Alcohol syndrome (FAS) is a condition occurring in some children of mothers who have consumed alcohol during pregnancy. It is characterized by physical and mental growth retardation, craniofacial anomalies and derangements of numerous body systems. A particular distressing feature of FAS is affection of the nervous system, which manifest as motor and sensory derangements and neurobehavioral deficits. However, no specific changes in the brain and spinal cord that could conclusively explain these neuronal defects have been reported. The aim of this study was to investigate the effects of a single high dose of ethanol exposure during early gestation on the peripheral nerve, with a view to understanding the neuronal disorders appearing in FAS. A chick model of FAS was used and embryos were exposed to 5%, 10% and 15% ethanol immediately after hatching. The effects on general growth and development and on the peripheral nerve of ethanol-exposed embryos were examined following the full period of gestation. There was a significant reduction in crown-rump length, head circumference and body weight in ethanol-exposed chicks as compared with appropriate control groups. The degree of growth retardation among the different alcohol exposed groups was dose dependent and significant prenatal embryo mortality was seen in those exposed to 15% ethanol. In embryos that survived the exposure to 15% ethanol, features of myelin degeneration were observed in the peripheral nerve in the majority of cases. No features of myelin degeneration were observed in embryos exposed to 5% and 10% ethanol or in the control groups. This study demonstrates the direct toxic effects of a single high dose of ethanol on developing embryos in general and the peripheral nerves in particular. Accordingly the pathological changes observed in peripheral nerves could account for some of the neurobehavioral deficits observed in FAS.

Effects of Genetic and Procedural Variation on Measurement of Alcohol Sensitivity in Mouse Inbred Strains

Mice from eight inbred strains were studied for their acute sensitivity to ethanol as indexed by the degree of hypothermia (HT), indexed as the reduction from pre-injection baseline of their body temperature. Two weeks later, mice were tested for their loss of righting reflex (LRR) after a higher dose of ethanol. The LRR was tested using the "classical" method of watching for recovery in animals placed on their backs in a V-shaped trough and recording duration of LRR. In a separate test, naive animals of the same strains were tested for HT repeatedly to assess the development of rapid (RTOL) and chronic tolerance (CTOL). We have recently developed a new method for testing LRR that leads to a substantial increase in the sensitivity of the test. Strains also have been found to differ in the new LRR test, as well as in the development of acute functional tolerance (AFT) to this response. In addition, our laboratory has periodically published strain difference data on the older versions of the HT and LRR responses. The earlier tests used some of the exact substrains tested currently, while for some strains, different substrains (usually, Nih versus Jax) were tested. We examined correlations of strain means to see whether patterns of strain differences were stable across time and across different test variants assessing the same behavioral construct. HT strain sensitivity scores were generally highly correlated across a 10-23 years period and test variants. The CTOL to HT was well-correlated across studies, and was also genetically similar to RTOL. The AFT, however, was related to neither RTOL nor CTOL, although this may be because different phenotypic end points were compared. The LRR data, which included a variant of the classical test, were not as stable. Measures of LRR onset were reasonably well correlated, as were those taken at recovery (e.g., duration). However, the two types of measures of LRR sensitivity to ethanol appear to be tapping traits that differ genetically. Also, the pattern of genetic correlation between HT and LRR initially reported in 1983 was not seen in current and contemporaneous studies. In certain instances, substrain seems to matter little, while in others, substrains differed a great deal. These data are generally encouraging about the stability of genetic differences.

CYP2E1 and catalase influence ethanol sensitivity in the central nervous system

Genetic factors are known to influence the sensitivity and tolerance to ethanol in humans and laboratory animals. Ethanol is metabolized to acetaldehyde mainly by the alcohol dehydrogenase pathway (ADHs) and, to a lesser extent, by microsomal oxidization (CYP2E1) and the catalase-H2O2 system. In this study, we examined the role of CYP2E1 and catalase in ethanol metabolism and sensitivity, using transgenic knockout Cyp2e1(-/-) mice, acatalasemic (Cs/Cs) mice, double mutant Cyp2e1(-/-)/Cs/Cs mice and their respective wild-type counterparts 129/sv, C3H/HeJ, 129/sv X C3H/HeJ mice. Ethanol was administered to the mouse lines and ethanol pharmacokinetics and sleep times were evaluated. Although the rates of whole blood ethanol elimination following i.p. administration were found to be similar regardless of dose or genetic stock, Cs/Cs, Cyp2e1(-/-) and Cyp2e1(-/-)/Cs/Cs mice exhibited longer ethanol-induced sleep times, especially at higher ethanol doses. This infers that there is less acetaldehyde produced in the brains of these animals and is in opposition to the idea that increased acetaldehyde increases the actions of ethanol. The Cyp2e1(-/-) animals produced lower whole blood levels of acetaldehyde than wild-type controls; however, this difference was seen only at higher doses of ethanol. The amount of acetaldehyde produced following the incubation of ethanol with liver and brain microsomes was greater in tissues derived from 129/sv than in those from Cyp2e1(-/-) mice. Although the contribution of CYP2E1 and catalase in ethanol oxidation may be of little significance, these enzymes appear to play a significant role in ethanol sensitivity in the brain.

Ethanol-induced preferences or aversions as a function of age in preweanling rats.

The reinforcing value of a relatively high ethanol dose (3 g/kg) administered intragastrically was investigated in preweanling rats. Ethanol intoxication on Postnatal Days 7-8 increased ethanol intake and enhanced the drug's palatability when tested 3 days later. In contrast, intoxication on Days 10-11 decreased ethanol intake and increased aversive responses to the drug. When reducing the delay between intoxication and test to 24 hr, the taste aversion was enhanced, whereas the preference was unaffected. Results suggest an ontogenetic change in pups' perception of the reinforcing value of ethanol that coincides with the end of a sensitive period for learning preferences in rats younger than 9 days old (T. L. Roth & R. M. Sullivan, 2003; R. M. Sullivan, M. Landers, B. Yeaman, & D. A. Wilson, 2000).