Polychlorinated biphenyls and methylmercury act synergistically to reduce rat brain dopamine content in vitro.

Consumption of contaminated Great Lakes fish by pregnant women is associated with decreased birth weight and deficits in cognitive function in their infants and children. These fish contain many known and suspected anthropogenic neurotoxicants, making it difficult to determine which contaminant(s) are responsible for the observed deficits. We have undertaken a series of experiments to determine the relevant toxicants by comparing the neurotoxic effects of two of these contaminants--polychlorinated biphenyls (PCBs) and methylmercury (MeHg)--both of which are recognized neurotoxicants. Striatal punches obtained from adult rat brain were exposed to PCBs only, MeHg only, or the two in combination, and tissue and media concentrations of dopamine (DA) and its metabolites were determined by high performance liquid chromatography. Exposure to PCBs only reduced tissue DA and elevated media DA in a dose-dependent fashion. Exposure to MeHg only did not significantly affect either measure. However, when striatal punches were simultaneously exposed to PCBs and MeHg, there were significantly greater decreases in tissue DA concentrations and elevations in media DA than those caused by PCBs only, in the absence of changes in media lactate dehydrogenase concentrations. Elevations in both tissue and media 3, 4-dihydroxyphenylacetic acid concentrations were also observed. We suggest that the significant interactions between these two toxicants may be due to a common site of action (i.e., toxicant-induced increases in intracellular calcium and changes in second messenger systems) that influences DA function. The synergism between these contaminants suggests that future revisions of fish-consumption guidelines should consider contaminant interactions.     

Iron deficiency alters dopamine transporter functioning in rat striatum

Iron deficiency anemia in early life produces profound changes in both in vivo and in vitro evaluations of dopamine (DA) functioning. This study employed both behavioral and biochemical approaches to examine the biological bases of alterations in striatal DA metabolism seen in iron-deficient rats. The purpose was to determine whether the DA transporter (DAT) was functionally altered in postweaning iron deficiency. Male and female 21-d-old Sprague-Dawley rats (n = 40) were fed either an iron-deficient (ID) diet (3 mg Fe/kg diet) or a control (CN) diet (35 mg Fe/kg diet) for 4 wk before behavioral testing. Motor activity responses to graded doses (3.75-30 mg/kg body) of the DA uptake inhibitor, cocaine, were significantly blunted in iron-deficient rats with a 50% higher half-maximal effective dose (ED(50)) in both males and females (CN-female, 7.1 +/- 0.9 mg/kg; ID-female, 11.2 +/-1.3 mg/kg; CN-male, 12.0 +/- 0.7 mg/kg; and ID-male, 17.0 +/- 1.8 mg/kg). Radioligand binding assays with (3)H-1-(2-(diphenylmethoxy)-ethyl)-4-(3-phenylpropyl) piperazine ((3)H-GBR12935) demonstrated that iron deficiency did not alter the affinity of the ligand for the DAT but did significantly decrease the density of the transporter by 30% in caudate putamen and 20% in nucleus accumbens. Iron deficiency also significantly decreased (3)H-DA uptake into striatal synaptosomes, but did not affect release of DA with potassium chloride stimulation. These experiments provide supporting evidence that elevated levels of extracellular DA in the striatum of iron-deficient rats is likely to be the result of decreased DAT functioning and not increased rates of release.     

Simultaneous changes in striatal dopamine,serotonin, and metabolites after withdrawal seizures in rats from dependence on alcohol

Ethanol dependence was achieved in male, Long-Evans rats after 8 days on a balanced liquid diet that supplied 4.5% ethanol. After 1-h access to a solution of 10% ethanol (95%)/5% sucrose, the rats were deprived of food, water, and ethanol for 9 h. Following 30-s key jingling, about 80% of the animals exposed to ethanol experienced tonic-clonic seizures. Neurochemical analyses of striatal tissues revealed a significant (p < 0.05) increase in dopamine (DA) and a significant decrease in serotonin (5-HT) in the ethanol-exposed rats that had seizures compared to control rats. Homovaniilic acid concentrations of the ethanol-treated rats with seizures were significantly higher than the levels found in ethanol-treated animals that had experienced no seizures. Daily average ethanol intake of the rats that had seizures vs. those that did not was almost the same at 16 g/kg/day. The findings indicate that rats experiencing ethanol withdrawal-induced seizures manifest opposite alterations in dopaminergic and serotoninergic activity compared to controls. The present results do not reveal if the striatal changes are caused by ethanol rather than by the seizures.     

In vivo formation of salsolinol induced by high acetaldehyde concentration in rat striatum employing microdialysis

AIMS: The in vivo formation of salsolinol (1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquionoline), an endogeneous condensation product of dopamine (DA) with acetaldehyde (AcH), was examined following the administration of cyanamide (CY) plus ethanol (EtOH) using microdialysis-high-performance liquid chromatography with electrochemical detection. METHODS: After the insertion of a microdialysis probe into the striatum, rats were treated with CY (a potent inhibitor of aldehyde dehydrogenase, 50 mg/kg), 4-methylpyrazole (4-MP, a strong inhibitor of alcohol dehydrogenase, 82 mg/kg), and CY + 4-MP, followed 1 h later by EtOH (1 g/kg), CY and 4-MP only by intraperitoneal administration. RESULTS: In the CY + EtOH group, salsolinol was detected in striatal dialysates and high AcH concentrations were found in the blood. The time course of changes in salsolinol concentrations correlated with blood AcH concentrations. In the other experimental groups, salsolinol in the dialysates and high AcH concentrations in the blood were not detected. CONCLUSIONS: These observations indicate that: (1) high AcH concentrations induce the formation of salsolinol in the rat striatum; (2) there is no effect of EtOH or AcH on striatal dialysate concentrations of DA and 5-hydroxytryptamine.     

Time course reversibility of liver and blood changes induced in weanling rats after polychlorinated biphenyl exposure during lactation

Phenoclor^® DP5 (DP5), a mixture of polychlorinated biphenyls (PCBs), was administered to lactating rat dams, every two days from postnatal days 2 to 20, at 50 mg/kg of body weight. Resulting effects on tissue concentrations of PCBs and on liver lipids, nucleic acids (DNA and RNA), proteins and blood lipids of offspring, at different times after weaning, until postnatal day 100, were studied. On postnatal day 21, DP5 contents found in liver, brain and fat of young rats indicated that a great part of the dam's body stores of PCBs was eliminated into the milk. These accumulations were related to an increase of relative liver weight, a decrease in DNA concentration and a rise in liver RNA, protein and phospholipid levels, indicating an inducing effect of DP5 in weanling rat. Changes in lipid metabolism were associated with increases of liver triacylglycerol and cholesterol and decrease of blood triacylglycerol. Both sexes exhibited an identical responsiveness to PCBs.The biochemical alterations observed in liver and blood at weaning, disappeared with time and were no longer detectable after postnatal day 40 in both sexes. Relative liver weight increase persisted until postnatal day 60 in males and postnatal day 100 in females. Residual aspects of alterations induced before weaning, by exposure via milk, could persist until postnatal day 100. Permanent effects of PCBs, after perinatal exposure could be assessed in later life of rats, when PCBs were almost completely cleared from body tissues.     

The alterations in neurotransmitters and their metabolites in discrete brain regions in the rats after inhalation of disinfectant, glutaraldehyde or ortho-phthalaldehyde for 4 weeks

Glutaraldehyde (GA) and ortho-phtalaldehyde (OPA) have been widely used as major components of disinfectants in hospitals. We evaluated the alterations in GA or OPA in rats after subacute inhalation exposure by determining levels of neurotransmitters (norepinephrine [NE], dopamine [DA], DA metabolites, dihydroxyphenylacetic acid [DOPAC] and homovanillic acid [HVA], indoleamine serotonin [5-HT] and 5-HT metabolite, 5-hydroxyindoleacetic acid [5-HIAA]) in discrete brain regions using high performance liquid chromatography (HPLC) equipped with an electrochemical detector. Female Wistar rats were exposed to 0, 50, 100, or 200 ppb gaseous GA or OPA by inhalation for 1 h per day, 5 d per week for 4 wk. Following the exposure, the brain of each rat was removed and dissected into cerebrum, cerebellum, medulla oblongata, midbrain, corpus striatum and hypothalamus. The neurotransmitters and their metabolites were extracted from each brain region, and determined by HPLC. Regarding GA, the daily water intake of the 50 or the 200 ppb exposed groups was significantly lower than that of the control. DA and 5-HIAA levels in the medulla oblongata among the GA exposed groups were significantly lower than those of the control. For OPA, the mean final body weight and daily food intake of the 100 or 200 ppb exposed groups were significantly lower than those of the control. The mean DA concentrations in the cerebrum in the groups exposed to OPA were significantly lower than those of the control. OPA may modulate DA metabolism in the cerebrum of female rats. The levels GA or OPA that induced alienations in neurotransmitters were comparable to those levels usually found in hospitals, further studies are warranted to evaluate the of safety of disinfectants containing GA or OPA.     

Cardiac catecholamine metabolism in copper-deficient rats

After parturition, Sprague-Dawley dams were fed diets containing either 0.6 (-Cu) or 6 (+Cu) mg of copper/kg of diet. Pups were weaned either to the diet of their dam or to the diet fed to dams in the other treatment group in a crossover design. At 7 wk of age, Cu-deficient rats were characterized by low tissue Cu and an enlarged heart with increased levels of dopamine (DA) and decreased levels of norepinephrine (NE). These changes resulting from Cu deficiency were independent of gender. In vivo synthesis of cardiac NE from DA in Cu-adequate rats was significantly greater than in Cu-deficient rats. Turnover of cardiac NE was estimated from the temporal change in the specific activity of [3H]NE. Fractional turnover rates (percentage per hour) of cardiac NE were similar in both dietary groups, although the cardiac turnover of NE (nanograms per hour) was 1.4-fold higher in Cu-adequate rats than in Cu-deficient rats. Repletion of Cu-deficient rats with dietary Cu increased the total amount of cardiac NE to 78 and 93% of control values after 1 and 2 d, respectively. Significant changes in the quantities of NE and DA in the heart of rats whose intake of Cu was restricted from birth were first detected at 4 wk of age, but cardiac hypertrophy was not observed until 5 wk of age. The data support the proposal that the altered levels of DA and NE in the heart of Cu-deficient rats are primarily the result of decreased activity of DA-B-monooxygenase and demonstrate that Cu supplementation rapidly repletes cardiac NE in Cu-deficient rats. Moreover, changes in the concentrations of NE and DA in the heart precede and may contribute to the development of cardiac hypertrophy.     

In vivo effects of inorganic mercury (HgCl(2)) on striatal dopaminergic system

In the present study, the effects of intrastriatal administration of different concentrations (40 microM, 400 microM, and 4 mM) of inorganic mercury (HgCl(2)) on the dopaminergic system of rat striatum were evaluated, using a microdialysis technique coupled to liquid chromatography-electrochemical detection. In previous studies, we discussed the effects of organic mercury (MeHg) administration on the striatal dopaminergic system on the basis of changes in the release and metabolism of striatal dopamine (DA). In the present study it is demonstrated that intrastriatal administration of all concentrations of HgCl(2) produced significant increases in the output of DA (1240, 2500, and 2658% for the concentrations of 40 microM, 400 microM, and 4 mM HgCl(2), respectively) from rat striatal tissue, associated with significant decreases in striatal levels of its metabolites dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) with the concentrations of 400 microM and 4 mM HgCl(2) (74.4 and 3.4% for DOPAC and 71.0 and 50.6% for HVA, respectively), whereas no changes in metabolite levels were observed with the concentration of 40 microM HgCl(2). These effects are explained as a result of stimulated DA release and/or changed DA metabolism. The effects of intrastriatal administration of HgCl(2) were compared with those of MeHg on DA extracellular levels.     

Urinary catecholamines in iron-deficient rats at rest and following surgical stress

The purpose of this study was to determine catecholamine concentrations both at rest and in response to a surgical stress in iron-deficient and control rats. Twenty-one-day-old rats were randomized to one of two groups which received a diet containing either 6 or 50 mg iron/kg. Three to five days later, when anemia was first detectable, urinary norepinephrine (NE) concentrations were already significantly elevated in the iron-deficient compared to control rats. In contrast, urinary dopamine (DA) became depressed after 10 days of the iron-deficient regimen. At 38 days of age, both groups were subjected to a surgical stress. NE and DA became elevated over baseline values in both diet groups during the 24-h period following surgery; NE remained significantly higher and DA significantly lower in the iron-deficient than in the control group. We conclude that changes in urine catecholamine concentration occur early in the development of iron deficiency and that they are characteristic of both baseline and stress conditions.     

Effects of occupational exposure to polychlorinated biphenyls on urinary metabolites of neurotransmitters: A cross-sectional and longitudinal perspective

BackgroundPolychlorinated biphenyls (PCBs) are chemicals which were used for industrial purposes and are known to induce various adverse health effects. They are also known to be neurotoxic and numerous targets within the central nervous system have been identified in previous studies. Specifically, the neurotransmitters dopamine (DA) and norepinephrine (NE) are influenced by PCBs as indicated in studies involving animals. However, limited evidence has been published documenting PCB induced changes in the neurotransmitter system in humans.ObjectiveIn the present study, we examined the association between a higher PCB body burden following occupational exposure and possible changes in human neurotransmitter metabolites.MethodsWithin a medical surveillance programme called HELPcB (Health Effects in High-Level Exposure to PCB) that monitors adverse health effects of occupational PCB exposure, urine samples were obtained (n_T1 = 166; n_T2 = 177 and n_T3 = 141). The urinary concentrations of the metabolites homovanillic acid (HVA; for DA) and vanillylmandelic acid (VMA; for NE) were analyzed. Blood samples were obtained by vena puncture in order to determine the internal exposure to PCBs with human biomonitoring.ResultsA cross-sectional analysis indicated a significant negative effect of PCB exposure on HVA and VMA. Longitudinally, an initially higher exposure to higher chlorinated PCBs was followed by constant reduced HVA level over three consecutive years. Exploratory analyses show different long-term effects for different PCBs according to their chlorination degree. A higher exposure with lower chlorinated PCBs leads to an increase of VMA and HVA. Conversely, a higher exposure to all PCBs results in a reduction of HVA.ConclusionThis study, to our knowledge, is the first to document changes in neurotransmitter metabolites after occupational PCB exposure in humans. This finding advances evidence obtained from past research, and identifies one potential pathomechanism in the central dopaminergic system of humans.     

Effects of polychlorinated biphenyls on the nervous system.

The neurological effects of polychlorinated biphenyls (PCBs) have been extensively investigated in humans and in animals. The main focus in human studies has been on the effects in neonates and young children, although studies of adults have also been conducted. A great deal of concern exists that even low levels of PCBs transferred to the fetus across the placenta may induce long-lasting neurological damage. Because PCBs are lipophilic substances, there is also concem that significant amounts might be transferred to nursing infants via breast milk. Studies in humans who consumed large amounts of Great Lakes fish contaminated with environmentally persistent chemicals, including PCBs. have provided evidence that PCBs are important contributors to subtle neurobehavioral alterations observed in newborn children and that some of these alterations persist during childhood. Some consistent observations at birth have been motor immaturity and hyporeflexia and lower psychomotor scores between 6 months and 2 years old. There is preliminary evidence that highly chlorinated PCB congeners, which accumulate in certain fish, are associated with neurobehavioral alterations seen in some newbom children. Subtle neurobehavioral alterations have also been observed in children bom to mothers in the general population with the highest PCB body burdens. Because of the limitations of epidemiological studies, these effects cannot be attributed entirely to PCB exposure. In one general population study, there was strong evidence that dioxins, as well as PCBs, were contributors to the neurobehavioral effects seen in exposed children. Children born to women who accidentally consumed rice oil contaminated with relatively high amounts of PCBs and chlorinated dibenzofurans (CDFs) during pregnancy also had neurodevelopmental changes. Studies in animals support the human data. Neurobehavioral alterations have been also observed in rats and monkeys following prenatal and/or postnatal exposure to commercial Aroclor mixtures, defined experimental congener mixtures, single PCB congeners, and Great Lakes contaminated fish. In addition, monkeys exposed postnatally to PCB mixtures of congeneric composition and concentration similar to that found in human breast milk showed learning deficits long after exposure had ceased. A few other generalizations can be made from the data in animals. It appears that ortho-substituted PCB congeners are more active than coplanar PCBs in modifying cognitive processes. In addition, one effect observed in both rats and monkeys--deficits on delayed spatial alternation--has been known to be induced by exposure to ortho-substituted PCBs, defined experimental mixtures, and commercial Aroclors. Both dioxin-like and non-dioxin-like PCB congeners have been shown to induce neurobehavioral alterations in animals. Changes in levels of neurotransmitters in various brain areas have also been observed in monkeys, rats, and mice. Of all the observed changes, the most consistent has been a decrease in dopamine content in basal ganglia and prefrontal cortex, but further research is needed before specific neurobehavioral deficits can be correlated with PCB-induced changes in specific neurotransmitters in specific brain areas.     

乐果对大鼠大脑分区多巴胺神经递质的影响

目的观察乐果染毒对大鼠大脑分区单胺类神经递质多巴胺(DA)及其代谢物(DOPAC)浓度变化的影响。方法104只雄性SD大鼠随机分为对照组(生理盐水)和乐果染毒低(38.9mg/kg)、中(83.7mg/kg)、高(180.0mg/kg)3个剂量组,腹腔一次注射染毒,给药后0.5、2、8和24h断头处死并分离脑组织(大脑皮层、纹状体、海马、脑干和小脑5部分)。用正丁醇、正庚烷、高氯酸、三氯甲烷处理脑组织样品,用高效液相色谱电化学检测法检测。结果不同时程的染毒组(低、中、高剂量组)大脑DA浓度及其代谢物DOPAC与对照组相比差异均有显著性(P<0.05)。DA及其代谢物DOPAC在纹状体分别增加了28%~122%、89%~538%;DA在其它分区变化差异没有显著性。DOPAC在大脑皮层有的剂量和时程增加了50%~72%、有的剂量和时程减少了31%~51%;在海马差异没有显著性;在脑干有的剂量和时程增加了4%~102%、有的剂量和时程减少了10%~16%;在小脑有的剂量和时程增加了6%~39%、有的剂量和时程减少了6%~23%。DA和DOPAC浓度在不同时程的低、中、高剂量组间差异均有显著性(P<0.05)。结论DA和DOPAC浓度有随染毒剂量和时程的增加而增加的趋势,存在剂量-效应和时程-效应关系。因此乐果的中毒存在非胆碱能机制,多巴胺类能机制。     

Immunologic biomarkers in relation to exposure markers of PCBs and dioxins in Flemish adolescents (Belgium)

In this study, we investigated 17- to 18-year-old boys and girls to determine whether changes in humoral or cellular immunity or respiratory complaints were related to blood serum levels of polychlorinated biphenyls (PCBs) and dioxin-like compounds after lifetime exposure in Flanders (Belgium). We obtained blood samples from and administered questionnaires to 200 adolescents recruited from a rural area and two urban suburbs. Physicians recorded medical history and respiratory diseases. We measured immunologic biomarkers such as differential blood cell counts, lymphocyte phenotypes, and serum immunoglobulins. As biomarkers of exposure, we determined the serum concentrations of PCBs (PCB 138, PCB 153, and PCB 180) and dioxin-like compounds [chemical-activated luciferase expression (CALUX) bioassay]. The percentages of eosinophils and natural killer cells in blood were negatively correlated with CALUX toxic equivalents (TEQs) in serum (p = 0.009 and p = 0.05, respectively). Increased serum CALUX TEQs resulted in an increase in serum IgA levels (p = 0.05). Furthermore, levels of specific IgEs (measured by radioallergosorbent tests) of cat dander, house dust mite, and grass pollen were also significantly and negatively associated with the CALUX TEQ, with odds ratios (ORs) equal to 0.63 [95% confidence interval (CI), 0.42-0.96], 0.68 (0.5-0.93), and 0.70 (0.52-0.95), respectively. In addition, reported allergies of the upper airways and past use of antiallergic drugs were negatively associated with CALUX TEQs, with ORs equal to 0.66 (0.47-0.93) and 0.58 (0.39-0.85), respectively. We found a negative association between IgGs and marker PCBs in serum (p = 0.009). This study shows that immunologic measurements and respiratory complaints in adolescents were associated with environmental exposure to polyhalogenated aromatic hydrocarbons (PHAHs). The negative correlation between PHAHs and allergic responses in adolescents suggested that exposure may entail alterations in the immune status.     

妊娠期甲基汞暴露对大鼠的母体毒性

目的　探讨妊娠期甲基汞暴露对大鼠的母体毒性效应。方法　用不同剂量甲基汞〔0 .0 0、0 .0 1、0 .0 5、2 .0 0mg/ (kg·d)〕于 Wistar大鼠孕 6～ 9天时连续 4天灌胃染毒 ,观察母鼠妊娠期、哺乳期的各种生理、生育过程 ,脑组织形态学和单胺类神经递质 (去甲肾上腺素、多巴胺、5 -羟色胺 )等指标。结果　与对照组相比 ,暴露组母鼠脑组织中单胺类神经递质的含量明显增高 (P<0 .0 1) ,呈现出剂量 -效应关系 (等级相关系数 rs分别为 0 .712 ,0 .90 3,0 .92 6 ) ,P均 <0 .0 1;其余指标未见明显异常。结论　本实验剂量下 ,妊娠期甲基汞暴露对大鼠的母体毒性尽管十分微弱 ,但母体脑组织的化学损伤已经发生。     

Age related differences in dopamine-stimulated adenylate cyclase sensitivity to "in vivo" chronic ethanol treatment

The effect of chronic ethanol consumption on adenylate cyclase activity was measured in striatal membranes derived from aged male rats. The results indicate that the cyclic AMP generating system of old rats has a different sensitivity to ethanol effect compared to the adult animals. In young animals the basal adenylate cyclase activity was enhanced by alcohol consumption while the DA stimulated cyclic AMP production was reduced. In contrast, in 24 months old rats ethanol reduced the basal adenylyl cyclase and enhanced the response to DA indicating a supersensitivity of adenylate cyclase linked DA receptors. This observation was further supported by 3H-Spiperone binding studies. In fact, a higher Bmax was measured in striatal membranes of aged ethanol-dependent rats in comparison to control.     

Alterations in dopamine metabolism by intraperitoneal ethanol in rats selected for high and low ethanol preference: A 3-methoxytyramine study

Effects of an ethanol dose (1 g/kg, IP) on the metabolism of dopamine (DA) in the nucleus accumbens, striatum and hypothalamus of ethanol-naive alcohol-preferring (AA) and alcohol-avoiding (ANA) rats were studied. Rats were sacrificed by focused-beam microwave irradiation of the brain 20 minutes after ethanol administration, and the concentrations of 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), assumed to reflect DA metabolism, and of 3-methoxytyramine (3-MT), assumed to reflect DA release, were measured using gas chromatography-mass spectrometry. Basal striatal DOPAC and HVA concentrations were higher in the AA rats in comparison with ANA rats. Ethanol increased HVA, but not DOPAC, concentration in the nucleus accumbens and striatum, but not in the hypothalamus. There was a significant rat line × ethanol treatment interaction with respect to HVA concentration in the nucleus accumbens. The increase in HVA was higher in the AA than ANA rats. Basal 3-MT concentration was not changed by ethanol, except in the nucleus accumbens, where a significant rat line × ethanol treatment interaction was found. A decrease in 3-MT concentration was only detected in the ANA rats. After inhibition of monoamine oxidase with pargyline hydrochloride (75 mg/kg, IP, 10 min before sacrifice), 3-MT accumulation was decreased by ethanol, especially in the nucleus accumbens of both AA and ANA rat lines as well as in that of nonselected Wistar rats. The results suggest that 1) DA metabolism to DOPAC and HVA is dissociable from DA release as reflected by 3-MT production, 2) ethanol, if anything, reduces DA release, and that 3) the AA and ANA rats differ in their basal DA metabolism and in the ethanol effects thereupon, but not in ethanol-induced changes in DA release.     

Effects of adolescent ethanol exposure on sleep in adult rats

Although adolescent ethanol (EtOH) exposure has been associated with long-lasting changes in brain function, little is known as to whether EtOH exposure during adolescence alters sleep and cortical arousal. This study examined protracted alterations in sleep in adult rats exposed to EtOH during adolescence. Adolescent male Wistar rats were exposed to EtOH vapor for 12 h/day for 5 weeks. Cortical electroencephalograms were obtained during 4-h recording sessions after 5 weeks of withdrawal from EtOH. Adolescent EtOH exposure significantly reduced the mean duration of slow-wave sleep (SWS) episodes and the total amount of time spent in SWS in EtOH-exposed rats, compared to controls. Spectral analysis revealed that adolescent EtOH exposure significantly increased cortical peak frequencies during SWS in the 2-4, 4-6, and 6-8 Hz bands. Taken together, our findings suggest that chronic EtOH exposure in adolescent rats reduces measures of SWS, an effect also seen as part of normal aging. Although the cellular and molecular mechanisms mediating the consequences of EtOH exposure on the aging process are not known, the similarities between adolescent EtOH exposure and aging merits further investigation.     

Second generation effects of maternal ethanol consumption on immunity to Trichinella spiralis in female rats.

The deleterious effects of maternal ethanol consumption on neonatal immune development and early immune responses has been well documented. However, the effects of such neonatal exposure to maternally consumed ethanol on the neonates' immune responses in their adult life, especially in combination with additional ethanol exposure, has received little attention. For these experiments, female rats were fed on either 6% ethanol or pair-fed isocaloric control Lieber-DeCarli liquid diets for 30 days prior to, and during, pregnancy and lactation. One day after weaning their pups, the mothers were infected with 1000 Trichinella spiralis larvae, and maintained on diets for an additional 20 days. At this time, they were challenged with 2000 T. spiralis larvae, killed 3 days later, and their immune status determined. These animals served as the first generation alcohol animals. Their female offspring served as the experimental second generation animals. These animals received maternal ethanol during pregnancy and lactation and control diet during their juvenile period (from weaning to 90 days of age). They were then subjected to a schedule of ethanol or pairfeeding, identical to the first generation dams. Two groups of second generation animals were established: Group 1 was exposed to ethanol during their dam's pregnancy and lactation periods only, with no subsequent ethanol treatment; Group 2 received ethanol during their dam's pregnancy and lactation periods and then again throughout their adult experimental period. Our previous studies showed only minimal changes following a secondary challenge in T. spiralis-immunized rats; however, neonates born to alcohol-consuming mothers did show some depressed secondary immune responses when challenged soon after weaning. We chose to use a secondary immune challenge to assess further immune alterations in second generation adult animals. No differences between any of the ethanol and pair-fed groups were observed in intestinal worm burdens, which is similar to data previously reported for adult alcohol-consuming animals. However, second generation group 2 animals demonstrated significantly reduced proliferation responses to T. spiralis antigen and Concanavalin A (Con A) stimulation relative to the ethanol first generation and to the second generation Group 1 animals. This group also demonstrated significantly lower absorbencies in the ELISA assay for specific IgM and IgG anti-T. spiralis antibodies than the pair-fed, ethanol first and second generation Group 1 animals. The proportion of total T cells and cytotoxic T cells was significantly lower and the proportion of natural killer cells was elevated in both second generation ethanol Groups 1 and 2 relative to the ethanol first generation and pair-fed groups. In addition, Group 2 second generation animals showed significantly lower proportions of total leukocytes and T cells than Group 1 second generation animals. Although secondary immune responses to T. spiralis infection were not altered in rats exposed to ethanol only as adults, exposure to maternal ethanol does affect some specific immune responses in second generation adult life and maternal exposure may exert cumulative immune effects in concert with later consumption of ethanol by offspring born to alcoholic mothers.