Paternal and Maternal Alcohol Consumption: Effects on Offspring in Two Strains of Rats

Long-Evans and Sprague-Dawley male rats were given liquid alcohol diets containing 35%, 17.5%, or 0% ethanol-derived calories (EDC). The latter two groups were pair fed to the higher alcohol diet group. A fourth group received lab chow and water ad libitum to assess the role of paternal undernutrition associated with alcohol consumption. After three or four weeks of diet consumption, these males were bred to females of the same strain. Pregnant females were divided into similarly treated alcohol groups and were fed these diets beginning on gestation Day 8, thus creating a factorial study with strain, paternal, and maternal alcohol consumption as main factors. Paternal alcohol consumption was associated with decreased litter size, decreased testosterone levels, and a strain-related effect on offspring activity. Offspring activity decreased for those sired by 35% and 17.5% EDC Long-Evans fathers. Activity also decreased for offspring sired by 17.5% EDC Sprague-Dawley fathers but increased for those sired by 35% EDC fathers. Paternal alcohol consumption did not affect postnatal mortality or passive avoidance learning of offspring. Maternal alcohol consumption was associated with lower birth weights, lower offspring weights at weaning, increased postnatal mortality, and poorer passive avoidance learning. However, offspring activity was not affected. In a separate study, levels of alcohol in the testes were found to be somewhat, but not significantly, lower than blood alcohol levels. DNA taken from sperm of Long-Evans males consuming alcohol, migrated farther under pulsed field electrophoresis than DNA from control fathers, suggestive of an alcohol-related effect on sperm DNA.     

Paternal Alcohol Consumption: Effects on Ocular Response and Serum Antibody Response to Pseudomonas aeruginosa Infection in Offspring

Male Swiss-Webster mice that consumed liquid alcohol diets containing 25, 20, 15, 10, 5, or 0% ethanol-derived calories (EDC) for 7 weeks were bred to untreated females that were not exposed to alcohol diets. Males receiving the 20-0% EDC diets were pair-fed to those consuming the 25% EDC diet. At approximately 60 days of age, male offspring were challenged with 10(8) colony forming units of Pseudomonas aeruginosa onto the scarified cornea of one eye. The ocular response was then evaluated macroscopically for 4 weeks. Male offspring sired by alcohol-consuming fathers exhibited more severe ocular infection at 24 hr postinfection compared with mice sired by control fathers and, after 8 days postinfection, more corneas of the 25% EDC-sired progeny perforated than did the other groups. At 30 days after infection, serum immunoglobulin titers (IgM, IgG, IgA) specific to P. aeruginosa were determined by ELISA. Although the majority of the mice were unable to restore corneal clarity within 30 days postinfection, a strong serum IgG response was detected in pooled sera from those animals tested. Offspring were reinfected in the contralateral control eye at 30 days postprimary infection. Most control animals were able to restore corneal clarity in the contralateral control eye within 30 days postinfection, but less than a third of the alcohol-sired offspring did so. Again, the pooled sera that was tested indicated a strong humoral response, despite differences in corneal clarity. These studies indicate an increased susceptibility to infection and an impairment in restoration of corneal clarity in offspring sired by alcohol-consuming males, which does not appear to be mediated by serum antibody mechanisms.     

Voluntary Alcohol Consumption in BXD Recombinant Inbred Mice: Relationship to Alcohol Metabolism

Studies were initiated to characterize behaviorally and biochemically C57BL/6J and DBA/2J inbred mice, as well as BXD Recombinant Inbred (RI) strains derived from them. The C57BL/6J, DBA/2J, and 7 BXD RI strains were tested for voluntary alcohol consumption (VAC) by receiving 4 days of forced exposure to a 10% (w/v) solution of alcohol, followed by 3 weeks of free choice between water and 10% alcohol. Measures of VAC included the absolute intake of alcohol (g/kg), as well as alcohol preference. A wide range of VAC was displayed by the various BXD RI strains with a continuous (rather than bimodal) distribution, indicating that there is likely to be additive effects of several genes involved in regulating alcohol-related behaviors. Kinetic characteristics of aldehyde dehydrogenase and catalase in liver and brain of the C57BL/6J, DBA/2J, and BXD strains of mice were determined to test the hypothesis that the genetic regulation of the levels of alcohol-metabolizing enzymes mediate differences in VAC. Aldehyde dehydrogenase activity was determined spectrophotometrically by observing the change in absorption at 340 nm. Catalase activity was determined by measuring oxygen production with a Yellow Springs Biological Oxygen monitor and oxygen electrode. There was a strong negative relationship between VAC and brain catalase activity in the BXD RI and parental strains. These data suggest that RI strains are likely to be useful genetic models in the examination of quantitative trait loci controlling VAC and other responses to alcohol.     

Paternal Alcohol Exposure Affects Offspring Behavior but not Body or Organ Weights in Mice

Male mice consumed liquid alcohol diets containing 20, 10, or 0% ethanol-derived calories (EDC) for 56-61 days. Animals fed the 10 and 0% EDC diets were pair-fed to 20% EDC animals. A nontreated ad libitum group was included to assess the effects of pair-feeding. After treatment, males were bred to nontreated females. Litter size, birthweight, bodyweight at 21 or 55 days of age, and organ weights except for thymus were not affected by paternal alcohol ingestion. There was a dose-related decrease in activity at 20 and 24 days of age in an activity chamber and a dose-related decrease in serum testosterone levels at 55 days of age in male offspring sired by alcohol-consuming males. Offspring sired by males consuming the 20% EDC diet also required fewer trials to learn a passive avoidance task and had longer latencies to reach the choice point in a T maze.     

Beverage Effects on Patterns of Alcohol Consumption

In this paper an analysis is made of beverage preferences and their effect on alcohol consumption patterns. For this purpose we have used the 1993 Spanish National Household Health Survey conducted on members of the population aged 16 or over. Beer and spirits are consumed more frequently by young people and wine by older people. The most consumed daily drink is wine and beer on a weekly basis. Men always drank more frequently and consumed a greater number of drinks per occasion than women for the three types of drink analyzed. The study shows that beverage preference is an important factor in the characterization of alcohol use patterns.     

Piracetam Reduces Alcohol Withdrawal in Mice without Potentiating Alcohol Sedative Effects

Piracetam reduced the intensity of withdrawal behavior in mice that had been exposed to ethanol for 4 days. However, in contrast to other antiwithdrawal agents, piracetam did not potentiate the sedative effects of acute ethanol administration. While the mechanism of action of piracetam is unknown, these findings suggest that piracetam elicits its antiwithdrawal effects by a mechanism differing from that of other agents.     

Two Generations of Maternal Alcohol Consumption in Mice: Effect on Pregnancy Outcome

This study investigated whether female offspring of alcohol-treated mothers are, themselves, more or less susceptible than control offspring to the deleterious effects of alcohol on the outcome of their own pregnancy. One group of pregnant C57BL mice was fed a liquid diet containing 25% ethanol-derived calories (EDC) and another group was pair-fed an isocaloric (0% EDC) control diet. A third group was fed lab chow ad libitum (LC). The female offspring resulting from those matings were subsequently mated upon reaching 90 days of age. These pregnant mice were then separated into three prenatal treatments (25% EDC, 0% EDC, and LC). On gestation-day 19, second generation fetuses were removed by cesarean section, weighed, and sexed. Results indicated that number of implants, live births, and percent prenatal mortality did not differ between groups. However, fetal weight was lower in groups prenatally exposed to ethanol than in controls, regardless of the prenatal history of the mothers, themselves. More importantly, the data suggest that offspring of alcohol-treated mothers who do not consume alcohol themselves during their own pregnancy may still have a tendency to have offspring of lower birth weight. On the other hand, if mothers prenatally exposed to alcohol do consume alcohol during their own pregnancy, the impact of fetal weight suppression is even greater than expected for in utero alcohol exposure alone. These effects may be due to the fact that mothers who were prenatally exposed to alcohol weighed less than controls at the time of becoming pregnant.(ABSTRACT TRUNCATED AT 250 WORDS)     

Long-term Effects of Alcohol Consumption in Male and Female Rats

Discrepancies exist regarding potential sex differences in the effects of ethanol on the myocardium. Therefore, the aims of this study were to determine if long-term ethanol consumption was associated with the development of a dilated cardiomyopathy (CM) in female rats and, second, to determine if the absence of ovarian hormones modulated this effect. Adult male and female (n = 6–8/group) sham-operated and ovariectomized (OVX) Sprague–Dawley rats received the Lieber DeCarli ethanol-containing (8% vol/vol) or control liquid diet for 8 months. All ethanol groups showed echocardiographic evidence of a cardiomyopathy; however, more significant ethanol-elicited differences were found in the male ethanol group than in either the female or female OVX groups. In addition, the male ethanol group had significant reductions in in vivo measures of contractility, such as the maximum derivative of change in systolic pressure and preload recruitable stroke work. Sex differences were also apparent in the pattern and degree of posterior and septal wall thickness changes, in that the male ethanol group had more posterior and septal wall thinning. In conclusion, similar to male rats long-term ethanol consumption in gonad-intact and OVX female rats is associated with the development of a dilated cardiomyopathy.     

Inhibition of Prostaglandin Synthesis by Indomethacin Does Not Affect Alcohol Consumption in Inbred Mice

The prostaglandin system has been implicated in mediating both the acute and chronic pharmacologic effects of alcohol. The effect of blockade of prostaglandin synthesis by indomethacin on genetically based alcohol preference in C57BL/6, C3H/He, and BALB/c mice was examined. Although strain typical alcohol preference patterns were observed, there was no effect of indomethacin on either naive or preestablished alcohol preference and consumption. Genetically transmitted alcohol preference may be a complex system in which prostaglandin synthesis does not play a part.     

Short-Term Effects of Maternal Alcohol Consumption on Lactational Performance

Previous research demonstrated that breast-feeding infants consumed significantly less milk during the immediate hours after their mothers consumed an acute dose of alcohol when compared with a nonalcoholic beverage. The present study tested the hypothesis that maternal alcohol consumption decreases the amount of milk available to the infant and alters milk composition in the short term. To this aim, 22 lactating women were tested on 2 days separated by 1 week; the women reported that they drank very little during pregnancy, but significantly increased alcohol intake during lactation. Each woman drank a 0.3 g/kg dose of alcohol in orange juice on one testing day and orange juice alone on the other; the order was counterbalanced. Immediately before drinking the beverage (baseline) and 2 hr after (postconsumption), women expressed their milk by using an electric breast pump until no milk had been secreted from either breast for 5 min. Although there was no difference in the energy content of the milk, maternal alcohol consumption slightly, but significantly, reduced the amount of milk produced by the lactating mother. These findings underscore the importance of determining whether and when infants compensate for the reductions in intake experienced at the breast following maternal alcohol consumption and how such changes impact on mother-infant interaction.     

Tension Reduction and the Effects of Prolonged Alcohol Consumption

It has been argued that the phenomenon of alcoholics experiencing mood deterioration during a heavy drinking bout discredits the Tension Reduction Hypothesis of alcohol consumption [6]. An experiment is described which sought to illustrate two ways in which this phenomenon and the hypothesis are compatible. Nine male alcoholics consumed 6gm of alcohol per kg bodyweight in 16 equal doses over two days. Ratings of mood and related physiological indices were used. In the long-term the physiological measures were elevated on the second drinking day and yet individual doses of alcohol lowered these throughout. Severely alcohol dependent subjects showed corresponding effects for subjective ratings ie long-term mood deterioration with short-term improvement. All subjects expected continuing to drink to be reinforcing relative to stopping. Thus alcohol may retain tension-reducing properties, and alcoholics may expect these, even if affective state was worsened overall during a drinking bout.     

Identification and Confirmation of Quantitative Trait Loci Regulating Alcohol Consumption in Congenic Strains of Mice

BACKGROUND: C57BL/6 inbred mice prefer alcohol whereas DBA/2 mice avoid it. We describe the construction of congenic strains of mice in which DBA/2 alleles for alcohol avoidance were placed on a C57BL/6 background using phenotypic selection. METHODS: Mice were primed to drink 10% v/v ethanol in water for 2 days before a two-bottle choice paradigm. N2 males who demonstrated an alcohol-avoidance phenotype were backcrossed to B6 females to construct 15 independent lines. RESULTS: Eight of these lines were lost due to failure to breed or absence of males with an alcohol-avoidance phenotype. The remaining sublines were split to form a total of 21 sublines. In the N7 and N9 generations, a genome scan located provisional quantitative trait loci (QTLs) on chromosomes 1, 2, 3, 6, and 9. Progeny testing confirmed QTLs on chromosomes 1 and 2. CONCLUSIONS: The QTL on chromosome 2 overlaps the 95% confidence interval of Alcp1 whereas that on chromosome 1 is new and has been called Alcp5. Marker-assisted selection was used in the N9 and subsequent generations to maintain the congenic lines and produce congenic strains.     

孕期酒精暴露致DNA甲基化异常对子代小鼠心脏发育相关基因表达的影响

目的:探讨DNA甲基化修饰失衡在孕期酒精暴露致子代小鼠心脏发育相关基因表达异常中的作用,为防治孕期饮酒所致的心脏发育畸形提供新思路。方法:选取24只健康昆明孕鼠按照随机数字表法等分为四组:正常组、对照组、酒精组、干预组。从孕期0.5 d^16.5 d,酒精组每日给予56%酒精(5 ml/kg)灌胃1次,干预组在上述酒精灌胃基础上给予每日1次腹腔注射DNA甲基转移酶(DNMT)抑制剂5-氮杂胞苷(2.5 mg/kg),对照组给予等量生理盐水灌胃和等量二甲基亚砜(DMSO)腹腔注射,正常组未予任何处理。于胎龄16.5 d收集各组子代小鼠心脏进行以下检测:(1)比色法检测DNMT活性;(2)甲基化测序检测心脏核心转录因子心肌细胞增强因子2A(MEF2A)启动子区CpG岛DNA甲基化水平,实时荧光定量PCR检测MEF2A转录水平;(3)染色质免疫共沉淀(ChIP)检测MEF2A对心脏结构基因心房利钠肽(ANP)、β肌球蛋白重链(β-MHC)和心肌肌钙蛋白T(cTnT)的调控作用;(4)蛋白免疫印迹法(Western blot)检测心脏结构基因ANP、β-MHC及cTnT的蛋白表达水平。结果:(1)比色法结果显示,酒精组及干预组胎鼠心肌组织中DNMT活性较对照组和正常组显著降低,差异有统计学意义(P<0.05);(2)心脏核心转录因子MEF2A启动子区CpG岛DNA甲基化水平在酒精组及干预组较对照组和正常组均显著降低(P<0.05);(3)实时荧光定量PCR结果显示,心脏核心转录因子MEF2A转录水平在酒精组及干预组较对照组和正常组显著升高(P<0.05);(4)ChIP结果表明,心脏核心转录因子MEF2A可结合心脏结构基因ANP、β-MHC及cTnT启动子区域直接参与上述基因的表达调控;(5)Western blot结果表明,酒精组和干预组小鼠心肌组织中ANP、β-MHC及cTnT的蛋白表达水平较对照组和正常组显著升高,差异有统计学意义(P均<0.05)。结论:酒精介导的DNA低甲基化可能参与了孕期酒精暴露所致的子代心脏发育相关基因表达异常。     

Effects of Chronic Alcohol Consumption on Hepatic Poly-ADP-Ribosylation in the Rat

Background: Poly-adenosine diphosphate (ADP)-ribosylation is involved in a variety of biological processes, which include DNA repair, malignant transformation, and apoptosis. It is of interest how this reaction is altered after long-term alcohol intake. Therefore, we determined long-term alcohol effects on hepatic poly-ADP-ribosylation in the rat.
Methods: Male Sprague Dawley® rats (four pairs) were pair-fed a nutritionally adequate liquid diet that contained ethanol as 36% of total energy and an isocaloric control diets for 4 weeks. Liver tissue homogenates and nuclear fractions were subjected to ADP-ribosylation with [³²P]nicotinamide adenine dinucleotide. The ADP-ribosylated proteins were separated by SDS-PAGE, followed by autoradiography. Expression of poly-ADP-ribose polymerase (PARP) also was evaluated by Western blotting.
Results: Incubation of rat liver homogenates in ADP-ribosylation reaction mixture resulted in a radiolabeling of a 116 kDa protein, most likely auto-ribosylation of PARP. This poly-ADP-ribosylation was increased significantly ( p < 0.025) after long-term alcohol intake. This alcohol effect was reproducible in nuclear fractions as well. Expression levels of PARP, however, were comparable between alcohol-fed rats and their pair-fed controls.
Conclusion: Poly-ADP-ribosylation, an important posttranslational modification of nuclear proteins, was increased significantly after chronic alcohol consumption in the rat.