Haughey, Heather M.; Kaiser, Alan L.; Johnson, Thomas E.; Bennett, Beth; Sikela, James M.; and Zahniser, Nancy R.  Norepinephrine transporter: A candidate gene for initial ethanol sensitivity in inbred Long-Sleep and Short-Sleep mice.  Alcoholism: Clinical & Experimental Research 29(10):1759-1768, October 2005.

Address correspondence to Heather M. Haughey, Department of Pharmacology, University of Colorado Health Sciences Center, Aurora, Colorado, USA. E-mail: heather.haughey@colorado.edu.

Summary: Molecular biological, neurochemical, and behavioral approaches were used to test the hypothesis that the norepinephrine transporter (NET) contributes to the differences in ethanol-induced loss of righting reflex (LORR) in inbred Long-Sleep (ILS) and inbred Short-Sleep (ISS) mice. The aim was to investigate the NET as a candidate gene contributing to this phenotype. The ILS and ISS mice carry different deoxyribonucleic acid haplotypes for NET, showing eight silent differences between allelic coding regions. Only the ILS haplotype is found in other mouse strains thus far sequenced. Brain regional analyses revealed that ILS mice have 30% to 50% lower [3H]NE uptake, NET binding, and NET messenger ribonucleic acid levels than ISS mice. Maximal [3H]NE uptake and NET number were reduced, with no change in affinity, in the ILS mice. These neurobiological changes were associated with significant influences on the behavioral phenotype of these mice, as demonstrated by (1) a differential response in the duration of ethanol-induced LORR in ILS and ISS mice pretreated with a NET inhibitor and (2) increased ethanol-induced LORR in LXS recombinant inbred (RI) strains, homozygous for ILS in the NET chromosomal region (44-47 cM), compared with ISS homozygous strains. This is the first report to suggest that the NET gene is one of many possible genetic factors influencing ethanol sensitivity in ILS, ISS, and LXS RI mouse strains. NIAAA Glossary Terms:  norepinephrine, transport proteins, ethanol, righting reflex, phenotype, genotype, gene expression, chromosome, selective breeding, animal strains, laboratory mice, DNA, haplotype, mRNA, chromosome, animal model

Alam, Imranul; Robling, Alexander G.; Weissing, Sarah; Carr, Lucinda G.; Lumeng, Lawrence; and Turner, Charles H.  Bone mass and strength: Phenotypic and genetic relationship to alcohol preference in P/NP and HAD/LAD rats.  Alcoholism: Clinical & Experimental Research 29(10):1769-1776, October 2005.

Address correspondence to Department of Orthopaedic Surgery, Indiana University School of Medicine, Indianapolis, Indiana 46202, USA.

Summary: The purpose was to determine whether there is a relationship between alcohol preference and high bone mass or strength and whether bone mass-regulating genes segregate during selective breeding of alcohol preferring rats. Six different lines of male rats with high or low preference for alcohol consumption were studied. The high alcohol preference lines were alcohol-preferring (P), high-alcohol-drinking 1 (HAD1), and high-alcohol-drinking 2 (HAD2). Their corresponding low alcohol preference lines were alcohol-nonpreferring (NP), low-alcohol-drinking 1 (LAD1), and low-alcohol-drinking 2 (LAD2). Bone mass phenotypes were determined using dual energy x-ray absorptiometry, peripheral quantitative computed tomography, and biomechanics in long bones and lumbar vertebrae from rats at 3 and 6 months of age. Bone mass and strength were significantly higher in P rats than in NP rats, mainly due to higher cortical bone in long bones and lumbar vertebrae. HAD2 rats also had significantly higher bone mass compared with LAD2 rats, but mostly due to increased trabecular bone leading to increased strength only in lumbar vertebra. Conversely, HAD1 rats had significantly lower bone mass and strength compared with LAD1 rats in long bones. Vertebral bone mass and strength did not differ between HAD1 and LAD1 rats. Thus there was no consistent relationship between preference for alcohol consumption and high bone mass or strength, as each alcohol-preferring rat line had its unique bone mass phenotypes. However, genes regulating bone mass and strength appear to segregate with alcohol preference genes in P and HAD rat lines, suggesting that alcohol preferring rat lines may be useful for identifying genes that regulate bone mass and structure. NIAAA Glossary Terms:  animal selectively bred for alcohol preference, animal strains, laboratory rat, bone mass density, phenotype, computed x-ray tomography, spine, animal model

Cowmeadow, R B.; Krishnan, H R.; and Atkinson, N S.  The slowpoke gene is necessary for rapid ethanol tolerance in Drosophila.  Alcoholism: Clinical & Experimental Research 29(10):1777-1786, October 2005.

Address correspondence to Section of Neurobiology and The Waggoner Center for Alcohol and Addiction Research, University of Texas at Austin, Austin, Texas 78712-0248, USA.

Summary: The slowpoke BK-type calcium-activated potassium channel gene has recently been shown to be involved in ethanol sensitivity in Caenorhabditis elegans and in rapid tolerance to the anesthetic benzyl alcohol in Drosophila. Drosophila mutants were used in this study to investigate the role of the slowpoke gene in rapid tolerance to sedation with ethanol vapor. Rapid tolerance was defined as a reduction in the sedative phase caused by a single previous sedation. The ethanol and water contents of flies were measured to determine if pharmacodynamic changes could account for tolerance. A saturated ethanol air stream caused sedation in <20 minutes and resulted in rapid tolerance that was apparent 4 hours after sedation. Two independently isolated null mutations in the slowpoke gene eliminated the capacity for tolerance. In addition, a third mutation that blocked expression specifically in the nervous system also blocked rapid tolerance. Water measurements showed that both ethanol and mock sedation caused equivalent dehydration. Furthermore, a single prior exposure to ethanol did not cause a change in the ethanol clearance rate. It was concluded that rapid tolerance, measured as a reduction in the duration of sedation, is a pharmacokinetic response to ethanol that does not occur without slowpoke expression in the nervous system in Drosophila. The slowpoke channel must be involved in triggering or producing a homeostatic mechanism that opposes the sedative effects of ethanol. NIAAA Glossary Terms:  gene, potassium channel, AOD sensitivity, ethanol, Drosophila melanogaster, AOD tolerance, mutation, gene expression, pharmacokinetics, homeostasis, sedative-hypnotics, animal study

Tsai, Chien-Sung; Loh, Shih-Hurng; Jin, Jong-Shiaw; Hong, Guo-Jieng; Lin, Hei-Ting; Chiung, Cheng-Shian; and Chang, Chung-Yi.  Effects of alcohol on intracellular pH regulators and electromechanical parameters in human myocardium.  Alcoholism: Clinical & Experimental Research 29(10):1787-1795, October 2005.

Address correspondence to Division of Cardiovascular Surgery, Tri-Service General Hospital, National Defense Medical Center, Neihu, Taipei, Taiwan, Republic of China. E-mail: sung1500@ndmctsgh.edu.tw.

Summary: Ethanol affects blood pressure and heart contractility and sometimes causes cardiac arrhythmia. This study assessed the effects of ethanol on intracellular pH (pHi) regulators and electromechanical parameters by superfusing various concentrations of ethanol into human myocardium obtained from hearts of patients undergoing corrective cardiac surgery. pHi was measured by a microspectrofluorimetry technique, while electrophysiological experiments were performed by traditional micropipette. Na+/H+ exchange (NHE) and Na+/HCO3– symporter (NHS) activities were measured after pHi recovery from intracellular acidosis induced by NH4Cl prepulse, while monocarboxylic acid transporter (MCT) activity was measured by a lactate adding/removing technique. It was shown in pHi experiments that ethanol could induce a biphasic, concentration-dependent (30–1000 mM) pHi change (i.e., alkalosis after acidosis) in human atrium in HEPES-buffered Tyrode solution. To a smaller extent, similar results were found when the superfusate was replaced by HCO3– buffered Tyrode solution. NHE activity was increased by a moderate concentration of ethanol (30 mM), but was inhibited in a concentration-dependent manner by higher concentrations of ethanol (>100 mM). In contrast, 30–1000 mM alcohol increased the activity of NHS in a concentration-dependent manner. Surprisingly, MCT activity was not affected by ethanol. In electromechanical experiments, ethanol (30-1000 mM) had a notable concentration-dependent inhibitory effect on the contractile force, while higher concentrations (>100 mM) decreased the action potential amplitude, upstroke velocity, duration of repolarization, and force of contractions in a concentration-dependent way. All these ethanol-induced pHi changes and electromechanical inhibitions were reversible. The authors believe that this study provides the first evidence that ethanol can affect pHi in human myocardial tissue by changing the activity of acid extruders (i.e., NHE and NHS). NIAAA Glossary Terms:  ethanol, pH, myocardium, transport proteins, acidosis, lactate, disorder of fluid or electrolyte or acid-base balance, heart function, muscle contraction, human study

Doremus, Tamara L.; Brunell, Steven C.; Rajendran, Pottayil; and Spear, Linda P.  Factors influencing elevated ethanol consumption in adolescent relative to adult rats.  Alcoholism: Clinical & Experimental Research 29(10):1796-1808, October 2005.

Address correspondence to Center for Developmental Psychology, Department of Psychology, Binghamton University, Binghamton, New York 13902-6000, USA.

Summary: Using a 24-hour, free-access, two-bottle-choice procedure between water and a sweetened solution with or without ethanol in nondeprived rats, this study examined the contribution of a variety of contextual and experimental variables (i.e., isolate-housing versus pair-housing, type of sipper tube, caloric value of solution, prior experimental perturbations) on alcohol consumption in both adolescent and adult Sprague-Dawley rats. Ethanol consumption was particularly magnified among adolescent rats using ball bearing-containing ball-point (BP) sipper tubes, with this exacerbated intake not due to caloric content of the ethanol solution. Isolation housing for 12 days did not alter ethanol consumption of adolescents relative to their socially housed counterparts, but did suppress consumption of isolated adults. An examination of differences in the relative magnitude of adolescent ethanol consumption across experiments in this series revealed that ethanol intake among adolescents was elevated not only by the inclusion of BP sipper tubes but also by staggering the timing of isolate housing relative to the presentation of the novel ethanol solution. These experiments demonstrate that adolescent animals consume significantly more ethanol than adult animals under a variety of home cage continuous-access circumstances, with the relatively greater intake of adolescents further magnified by a number of test conditions. Subtle experimental details often thought to be innocuous can have a substantial impact on overall amount of voluntary ethanol consumption observed in both adolescent and adult animals. NIAAA Glossary Terms:  ethanol, AOD consumption, adolescent, adult, laboratory rat, age differences, animal behavior, research issue, variable, characteristic, factor, animal study

Ristuccia, Robert C. and Spear, Linda P.  Sensitivity and tolerance to autonomic effects of ethanol in adolescent and adult rats during repeated vapor inhalation sessions.  Alcoholism: Clinical & Experimental Research 29(10):1809-1820, October 2005.

Address correspondence to Center for Developmental Psychobiology, Department of Psychology, Binghamton University, Binghamton, New York 13902-6000, USA.

Summary: A possible contributor to the increased ethanol consumption often seen during adolescence in humans and in various animal models is age differences in ethanol sensitivity and tolerance. This study examined the impact of age on ethanol-related alterations in the autonomic nervous system. Sensitivity to the initial ethanol challenge and chronic tolerance as well as acute and protracted withdrawal-like phenomena were assessed in male adolescent and adult Sprague-Dawley rats, using implanted telemetry probes with ethanol delivered by vapor inhalation. Adolescents and adults showed similar ethanol-induced tachycardia and activity suppression, but adolescents were more sensitive to the hypothermic effect of ethanol, contrary to other results from the authors' laboratory and elsewhere using intragastric or intraperitoneal ethanol administration. Although little tolerance to ethanol's tachycardic or activity suppressant effects was seen after repeated ethanol inhalation sessions, chronic tolerance to ethanol's hypothermic effect developed faster in adults than in adolescents. A withdrawal-like syndrome, characterized by bradycardia and hypoactivity, typically emerged during the dark phase of the diurnal cycle after ethanol vapor exposure sessions. These effects were observed in rats of both ages, with the bradycardic effect more pronounced in adolescents. In contrast to results indicating that adolescents may be less sensitive than adults to ethanol's hypothermic effect when ethanol is administered via bolus injection/intubation, adolescents appear more sensitive and develop tolerance to ethanol's hypothermic effects more slowly than adults when ethanol is administered at a more moderate rate by vapor inhalation. NIAAA Glossary Terms:  ethanol, inhalation, AOD consumption, AOD sensitivity, AOD tolerance, age differences, adolescent, adult, laboratory rat, tachycardia, bradycardia, hypothermia, chronic AODE, intragastric administration, intraperitoneal administration, physical activity, AOD withdrawal syndrome, animal study

Marinelli, Peter W.; Bai, Li; Quirion, Remi; and Gianoulakis, Christina.  A microdialysis profile of met-enkephalin release in the rat nucleus accumbens following alcohol administration.  Alcoholism: Clinical & Experimental Research 29(10):1821-1828, October 2005.

Address correspondence to Biobehavioural Pharmacology Section, Centre for Addiction and Mental Health, Toronto, Ontario, Canada.

Summary: The majority of studies of the role of the endogenous opioid system in mediating alcohol intake have concentrated on endorphinergic systems. Other opioid systems have received comparably less attention despite some compelling evidence that implicates enkephalinergic peptide systems, particularly met-enkephalin, in mediating alcohol preference. This study investigated the effect of alcohol administration on extracellular levels of met-enkephalin in the rat nucleus accumbens (NA), a brain region that plays a significant role in the processes underlying reinforcement and stress. Male Sprague-Dawley rats were implanted with a microdialysis probe aimed at the shell region of the NA. Artificial cerebrospinal fluid was pumped at a rate of 1.75 μl/min in awake and freely moving rats, and dialysates were collected at 30-minute intervals. After several baseline collections, rats were injected intraperitoneally with either physiological saline or one of four doses of ethanol: 0.8, 1.6, 2.4, or 3.2 g/kg body weight. Met-enkephalin levels in the dialysates were analyzed with solid-phase radioimmunoassay. Within the first 30 minutes of administration, an ethanol dose of 1.6 g/kg caused a significant and prolonged elevation in the extracellular levels of met-enkephalin. Ethanol did not have a major effect on the release of met-enkephalin at any other dose. Enkephalins may modulate local neurotransmitter release by binding to presynaptic δ-opioid receptors, or, they may inhibit effector cells by binding to postsynaptic δ- or μ-opioid receptors. This may be one of multiple neurological mechanisms that modulate alcohol-drinking behavior. NIAAA Glossary Terms:   endogenous opioids, ethanol, methionine enkephalin, neurotransmitters, nucleus accumbens, microdialysis, intraperitoneal administration, delta-opioid receptors, mu-opioid receptors, reinforcement, stress, AOD use behavior, animal behavior, controlled study, laboratory rat, animal study

Slawecki, Craig J. and Ehlers, Cindy L.  Enhanced prepulse inhibition following adolescent ethanol exposure in Sprague-Dawley rats.  Alcoholism: Clinical & Experimental Research 29(10):1829-1836, October 2005.

Address correspondence to Craig J. Slawecki, Scripps Research Institute, Department of Neuropharmacology, La Jolla, California 92037, USA. E-mail: cslawecki@scripps.edu.

Summary: Recent studies have demonstrated that ethanol differentially affects adolescents and adults. The objective of this study was to compare the effects of 2-week exposure to ethanol during adolescence or adulthood on the acoustic startle response (ASR) and prepulse inhibition (PPI). Male Sprague-Dawley rats were exposed to ethanol vapor 12 hours a day (from 6 pm to 6 am) for 14 days during adolescence or adulthood. The ASR and PPI were assessed 6 days after the cessation of ethanol vapor exposure. During ethanol treatment, overall blood alcohol levels averaged 230 to 250 mg/dl in the adolescent and adult treatment groups. Assessment of the ASR revealed that latency to startle was shorter in adolescents than in adults, but ASR latency was not altered by ethanol exposure. In addition, ASR magnitude was lower in adolescents and was decreased in ethanol-exposed rats on startle trials. Ethanol exposure significantly enhanced PPI, but only in adolescents. These results further demonstrate differential sensitivity of adolescents and adults to the effects of ethanol exposure. Specifically, a 2-week period of ethanol exposure during adolescence selectively enhanced PPI, a neurobehavioral index of sensorimotor gating. However, ASR magnitude was decreased by ethanol exposure regardless of age. On the basis of previous studies, the effects of ethanol exposure on PPI data could indicate that adolescent rats exposed to ethanol are more likely to exhibit behavioral inflexibility and that ethanol acts as a more potent physical stressor in adolescent rats. NIAAA Glossary Terms:  adolescent, adult, age differences, ethanol, inhalation, laboratory rat, animal behavior, comparative study, stressor, animal study

Rewal, Mridula; Wen, Yi; Wilson, Andrew; Simpkins, James W.; and Jung, Marianna E.  Role of parvalbumin in estrogen protection from ethanol withdrawal syndrome.  Alcoholism: Clinical & Experimental Research 29(10):1837-1844, October 2005.

Address correspondence to Mridula Rewal, Department of Pharmacology and Neuroscience, University of North Texas Health Science Center at Fort Worth, Fort Worth, Texas 76107-2699, USA. E-mail: mrewal@hsc.unt.edu.

Summary: Parvalbumin is a calcium-binding protein that has been implicated in protecting neurons from hyperexcitability by sequestering intracellular calcium. This study examined whether ethanol exposure or ethanol withdrawal (EW) alter the levels of parvalbumin in a manner that is protected by 17β-estradiol (E2). Ovariectomized rats implanted with E2 (EW/E2) or oil pellets (EW/Oil) received chronic ethanol (7.5% w/v, 5 weeks) or control dextrin (Dex/Oil and Dex/E2) diets. At 0 hours, 24 hours, and 2 weeks of ethanol withdrawal, three brain areas (cerebellum, hippocampus, and cortex) were prepared for immunoblotting and immunohistological assessment of parvalbumin. At 24 hours of EW, the EW/Oil group showed reduced levels of parvalbumin protein and parvalbumin-positive neurons in the cerebellum and hippocampus compared with the dextrin control and the EW/E2 groups. At 2 weeks of ethanol withdrawal, the reduced levels of parvalbumin persisted in the cerebellum but recovered toward the control levels in the hippocampus. The cortex showed no change in parvalbumin levels in any of the treatment groups. When tested at 24 hours of ethanol withdrawal, the magnitude of withdrawal signs inversely correlated with the levels of parvalbumin in the cerebellum and hippocampus. Ethanol exposure itself did not affect parvalbumin levels. These results suggest that ethanol withdrawal, rather than ethanol exposure, reduces parvalbumin levels in a manner that is brain region specific and is protected by estrogen. Disturbed parvalbumin homeostasis is hypothesized to play a role in the hyperexcitability of ethanol withdrawal signs. NIAAA Glossary Terms:   calcium binding protein, ethanol, AOD withdrawal syndrome, estradiol, estrogens, cerebellum, hippocampus, cerebral cortex, neuron, immunoblotting, histologic study, controlled study, laboratory rat, homeostasis, hyperexcitability, animal study

Allen, Gregg C.; West, James R.; Chen, Wei-Jung A.; and Earnest, David J.  Neonatal alcohol exposure permanently disrupts the circadian properties and photic entrainment of the activity rhythm in adult rats.  Alcoholism: Clinical & Experimental Research 29(10):1845-1852, October 2005.

Address correspondence to Texas A and M University System Health Science Center, College of Medicine, Department of Human Anatomy and Medical Neurobiology, College Station, Texas 77843-1114, USA.

Summary: The long-term effects of neonatal alcohol exposure on circadian behavioral activity were examined in adult rats. Artificially reared Sprague-Dawley rat pups were exposed to ethanol (4.5 g/kg/day) or isocaloric milk formula (gastrostomy control; GC) on postnatal days 4-9. At 2 months of age, rats from the ethanol, GC, and suckle control (SC) groups were housed individually, and properties of the circadian rhythm in wheel-running behavior were continuously analyzed during exposure to a 12-hour light:12-hour dark photoperiod (LD 12:12) or constant darkness (DD). Neonatal ethanol exposure had distinctive effects on the rhythmic properties and quantitative parameters of adult wheel-running behavior. Ethanol-treated rats were distinguished by unstable and altered entrainment to LD 12:12 such that their daily onsets of activity were highly variable and occurred at earlier times relative to control animals. In DD, circadian regulation of wheel-running behavior was altered by neonatal ethanol exposure such that the free-running period of the activity rhythm was shorter in ethanol-exposed rats than in control animals. Total amount of daily wheel-running activity in ethanol-treated rats was greater than that observed in the SC group. In addition, the circadian activity patterns of ethanol-exposed rats were fragmented such that the duration of the active phase and the number of activity bouts per day were increased. These results indicate that neonatal alcohol exposure produces permanent changes in the circadian regulation of the rat activity rhythm and its entrainment to LD cycles. These long-term alterations in circadian behavior, along with the developmental ethanol-induced changes in suprachiasmatic nucleus endogenous rhythmicity, may have important implications in clinical sleep-wake disturbances observed in neonates, children, and adults exposed to alcohol in utero. NIAAA Glossary Terms:   postnatal alcohol exposure, neonate, adulthood, circadian rhythm, animal behavior, laboratory rat, controlled study, sleep disorder, animal study

Javors, Martin A.; Ginsburg, Brett C.; Friesenhahn, Greg; Delallo, Leo; and Lamb, Richard J.  Rat breathalyzer.  Alcoholism: Clinical & Experimental Research 29(10):1853-1857, October 2005.

Address correspondence to Martin A. Javors, Department of Psychiatry, University of Texas Health Science Center, San Antonio, Texas 78229, USA. E-mail: javors@uthscsa.edu.

Summary: This article describes the estimation of blood alcohol concentration (BAC) in laboratory rats by measuring ethanol concentration in breath (BrAC) using a specialized apparatus in combination with a gas chromatography system. The apparatus consisted of a body chamber, a plastic cylinder, from which the head of the rat protruded, a head chamber, and a water-jacketed cylinder, in which the rat's head was placed while the breath sample was collected. The breath sample was withdrawn from the head chamber through a sample loop by a Minipuls pump and then injected directly into the gas chromatography system, which was equipped with a flame ionization detector for the quantification of ethanol. For these experiments, Lewis rats were catheterized 1 week before the commencement of the experiments so blood samples could be collected at exactly the same time as the breath samples. The results showed that Lewis rats can be trained to enter and be secured in the body chamber and that they appear to be comfortable for periods as long as 150 minutes. The pharmacokinetic curves for BrAC and BAC had essentially identical profiles. Cmax for BrAC and BAC at 8 minutes after the intraperitoneal injection of ethanol was directly proportional to the ethanol doses. The ratio of BrAC expressed as peak area to BAC (expressed as mM) was calculated to be 3282. This conversion factor can be used to directly estimate the BAC from the BrAC. It was concluded that the rat breathalyzer is an accurate and convenient laboratory method for noninvasive estimation of BAC. The procedure is expected to be particularly useful for studies requiring repeated assessment of alcohol levels. NIAAA Glossary Terms:  ethanol, BAC, BAC method, breath alcohol analysis, laboratory measurement, laboratory rat, gas chromatography, pharmacokinetics, intraperitoneal administration, correlation analysis, animal study

Moos, Rudolf H. and Moos, Bernice S.  Paths of entry into Alcoholics Anonymous: Consequences for participation and remission.  Alcoholism: Clinical & Experimental Research 29(10):1858-1868, October 2005.

Address correspondence to Rudolf H. Moos, Center for Health Care Evaluation, Department of Veterans Affairs and Stanford University, Palo Alto, California, USA. E-mail: rmoos@stanford.edu.

Summary: Three groups of individuals with alcohol use disorders who, in the first year after initiating help-seeking were compared: those who entered Alcoholics Anonymous (AA) only, those who entered professional treatment and AA together, and those who entered professional treatment only. A sample of initially untreated individuals (N = 362) was surveyed at baseline and 1 year, 3 years, 8 years, and 16 years later. At each contact point, participants described their participation in AA and treatment and their current alcohol-related functioning. They also described their reasons for entering AA and/or treatment and the perceived benefits of these sources of help. Compared with individuals who initially participated only in treatment but later entered AA, those who entered treatment and AA together participated in AA longer and more frequently and were more likely to achieve remission. Among individuals who initially participated only in AA, those who later entered treatment had poorer remission outcomes than those who did not enter treatment. Longer duration of participation in AA was associated with a higher likelihood of remission at all four follow-ups; individuals who dropped out of AA were more likely to relapse or remain nonremitted. In conclusion, compared with individuals who participated only in professional treatment in the first year after they initiated help-seeking, individuals who participated in both treatment and AA were more likely to achieve remission. Individuals who entered treatment but delayed participation in AA did not appear to obtain any additional benefit from AA. NIAAA Glossary Terms:  treatment method, AOD dependence, Alcoholics Anonymous, professional, combined modality therapy, treatment factors, treatment outcome, treatment research, comparative study, human study

Kuperman, Samuel; Chan, Grace; Kramer, John R; Bierut, Laura; Bucholz, Kathleen K.; Fox, Louis; Hesselbrock, Victor; Numberger, John I. Jr; Reich, Theodore; Reich, Wendy; and Schuckit, Marc A.  Relationship of age of first drink to child behavioral problems and family psychopathology.  Alcoholism: Clinical & Experimental Research 29(10):1869-1876, October 2005.

Address correspondence to Samuel Kuperman, Department of Psychiatry, University of Iowa Hospitals and Clinics, Iowa City, IA 52242-1057, USA. E-mail: samuel-kuperman@uiowa.edu.

Summary: The objective was to quantify the contributions of antecedent variables in four areas to prediction of early age of first drink (early AFD): child characteristics, family demographics, family psychopathology, and child behavior problems. Using data from a multicenter study on alcoholism, the authors first investigated the differences between two groups of children (ages 7–17 years), one from families heavily loaded for alcohol dependence and the other from population controls. Second, a multidomain, multistep regression model using child characteristics, family demographics, family psychopathology, and child behavior problems was performed to determine significant contributors to predicted AFD. Five variables initially contributed to the prediction of AFD. These included gender, age at interview, the number of adult sibs with alcohol dependence, being held back a year in school, and conduct scale score. However, the number of conduct symptoms appeared to contain the contributions of gender and being held back a grade in school, so these two variables were subsequently removed from the model. The remaining three variables explained 45% of the model variance; age at interview accounted for 38.3%, conduct scale score accounted for 6.2%, and the number of alcohol-dependent adult sibs accounted for 0.5%. No family history measures of alcohol dependence or antisocial personality disorder were contributory to the prediction model for AFD. Both the "number of conduct symptoms" and the "number of adult sibs with alcohol dependence" are inversely associated with predicted AFD. The latter variable appears marginally predictive of AFD and suggests a condition in which the child's household, regardless of strength of family history of alcoholism (or antisocial personality disorder), appears conducive to early drinking. Thus child and environmental factors are stronger predictors of AFD than family history. NIAAA Glossary Terms:   early AODU onset, underage drinking, predictive factor, risk factors, demographic characteristics, individual differences, family dysfunction, psychopathology, childhood behavioral problem, familial alcoholism, regression analysis, gender differences, age differences, family AODU history, conduct disorder, antisocial personality disorder, environmental factors, human study

Yokoyama, Akira; Yokoyama, Tetsuji; Kumagai, Yoshiya; Kato, Hoichi; Igaki, Hiroyasu; Tsujinaka, Toshimasa; Muto, Manabu; Omori, Tai; Yokoyama, Masako; and Watanabe, Hiroshi.  Mean corpuscular volume, alcohol flushing, and the predicted risk of squamous cell carcinoma of the esophagus in cancer-free Japanese men.  Alcoholism: Clinical & Experimental Research 29(10):1877-1883, October 2005.

Address correspondence to Akira Yokoyama, National Hospital Organization Kurihama Alcoholism Center, 5-3-1 Nobi, Yokosuka, Kanagawa, Japan. E-mail: a_yokoyama@kurihama1.hosp.go.jp.

Summary: Because some of the causes of increased mean corpuscular volume (MCV) and esophageal squamous cell carcinoma (ESCC), including alcoholism, acetaldehyde exposure, smoking, and poor nutrition are common to both, macrocytosis has been used as a predictor of early ESCC in Japanese alcoholics. This study examined whether this was also true in the Japanese general population by comparing the MCV of cancer-free Japanese men (N = 522) at high risk of ESCC as defined using drinking, smoking, dietary habits and aldehyde dehydrogenase-2 (ALDH2) genotype in a previous case-control study of ESCC involving them as control subjects. MCV was significantly correlated with ESCC risk predicted by drinking combined with ALDH2 genotype, smoking, or fruit intake. Men at higher risk of ESCC were more frequent in the groups with higher MCV (p < 0.0001 for trend). The replies to a questionnaire about facial flushing in response to alcohol showed that the trend was more prominent in men with current/former flushing, a surrogate marker for inactive ALDH2, than in men with no flushing (p < 0.0001). In comparison with the mean risk of men with MCV ≤ 93 fl (lowest quartile), that of current/former flushing men with MCV ≥ 99 fl (highest quartile) was 6.35-fold higher, whereas that of never-flushing men with MCV ≥ 99 fl was 2.50-fold higher. The sensitivity and specificity of the combination of moderate-to-heavy drinking and either MCV ≥ 99 fl or current/former flushing, either 30+ pack-years or MCV ≥ 99 fl or either 30+ pack-years or current/former flushing for detection of high-risk persons ranking in the top 10%, was 85% and 84%, 94% and 76%, or 98% and 77%, respectively. MCV and alcohol flushing may be useful for selecting candidates to screen for this high-mortality cancer in alcoholic and nonalcoholic Japanese men. NIAAA Glossary Terms:  cancer, esophageal disorder, squamous cell carcinoma, mean corpuscular volume, risk factors, risk analysis, AOD dependence, acetaldehyde, aldehyde dehydrogenases, genotype, smoking, diet, AOD consumption, alcohol flush reaction, biological markers, Japan, human study

McKee, Martin; Suzcs, Sandor; Sarvary, Attila; Adany, Roza; Kiryanov, Nikolay; Saburova, Ludmila; Tomkins, Susannah; Andreev, Evgeny; and Leon, David A.  The composition of surrogate alcohols consumed in Russia.  Alcoholism: Clinical & Experimental Research 29(10):1884-1888, October 2005.

Address correspondence to Martin McKee, European Centre on Health of Societies in Transition, London School of Hygiene and Tropical Medicine, London, United Kingdom. E-mail: martin.mckee@lshtm.ac.uk.

Summary: During a case-control study examining determinants of premature death among working age men in Russia, it became clear that a significant percentage of the population (7.3%) were drinking a variety of surrogate alcohol products (products not legally sold for consumption). In this population, where there is a high death rate from alcohol-related causes, including acute alcohol poisoning, it was important to know what these products contained. The identity of the products was identified from the survey of controls, and representative samples were obtained for composition analysis using gas chromatography and mass spectrometry. Three broad groups of products were identified: samogon (home-produced spirits); medicinal compounds; and other spirits (mainly sold as aftershave lotions). Commercially produced vodkas were used for comparison. Samogon contained lower quantities of ethanol than vodka (mean, 39 versus 44 volumetric percentage [v/v%]) but also contained certain toxic long-chain alcohols. Medicinal compounds contained only ethanol, at a higher concentration than vodka (mean, 66 v/v%), while the other spirits, which were also essentially pure ethanol, contained a mean of 94 v/v%. Thus a significant number of Russian men are drinking products that have either very high concentrations of ethanol or contaminants known to be toxic. These products are untaxed and thus much less expensive than vodka. The authors urge policy responses that target their production and consumption. NIAAA Glossary Terms:   alcohol product, nonbeverage alcohol, nonfood alcohol product, Russia, AODR mortality, AOD poisoning, acute AODE, survey, gas chromatography, spectrometry, distilled alcoholic beverage, product substitution, aliphatic alcohols, congeners, toxic substances, public policy on AOD, human study

Soardo, Giorgio; Donnini, Debora; Varutti, Rosanna; Moretti, Massimo; Milocco, Carla; Basan, Lorenza; Esposito, Walter; Casaccio, Daniele; Stel, Giuliana; Catena, Cristiana; Curcio, Francesco; and Sechi, Leonardo A.  Alcohol-induced endothelial changes are associated with oxidative stress and are rapidly reversed after withdrawal.  Alcoholism: Clinical & Experimental Research 29(10):1889-1898, October 2005.

Address correspondence to Giorgio Soardo, Department of Internal Medicine and Liver Unit, Institute of Clinical Pathology, University of Udine School of Medicine, Udine, Italy. E-mail: giorgio.soardo@med.uniud.it.

Summary: Although heavy alcohol drinkers are at increased risk of developing cardiovascular events, moderate alcohol intake is associated with reduced incidence of cardiovascular death. This paradox might reflect a dose-related effect of different alcohol intakes on endothelial function and this, in turn, might depend on changes in oxidative stress. This study tested the effects of alcohol withdrawal in heavy drinkers and compared the plasma levels of endothelin-1, nitric oxide, plasminogen activator inhibitor-1, von Willebrand factor, malondialdehyde, and intracellular glutathione with those of alcoholics who did not modify their alcohol intake and nondrinkers. The same parameters after withdrawal of ethanol exposure were assessed In human endothelial cells that had been cultured for 2 weeks in the presence of different concentrations of ethanol. Alcohol increased the levels of endothelin-1, nitric oxide, and plasminogen activator inhibitor-1 and decreased the levels of von Willebrand factor both in vivo and in vitro. These changes were dose dependent, rapidly reversed after withdrawal of exposure, and associated with the presence of increased oxidative stress as indicated by increased levels of both malondialdehyde and intracellular glutathione. Blockade of oxidative stress by incubation of endothelial cells in the presence of oxidant scavengers prevented the alcohol-induced functional modifications of the endothelium. In conclusion, alcohol affects endothelial function with an effect that is mediated by an activated oxidative stress and is rapidly reversed after withdrawal. Dose-related endothelial responses to different alcohol intakes might translate in either vascular protection or vascular damage. NIAAA Glossary Terms:  heavy AOD use, moderate AOD use, AOD dependence, cardiovascular disorder, risk factors, protective factors, protective drug effect, endothelium, endothelial cell, endothelin, nitric oxide, plasminogen activator, malondialdehyde, glutathione, oxidative stress, in vivo study, in vitro study, dose-response relationship, antioxidants, human study

Sumida, Ken D.; Cogger, Alma A.; Arimoto, Steven M.; and Matveyenko, Aleksey V.  Opposing effects of chronic alcohol consumption on hepatic gluconeogenesis for female versus male rats.  Alcoholism: Clinical & Experimental Research 29(10):1899-1905, October 2005.

Address correspondence to Ken D. Sumida, Department of Biological Sciences, Chapman University, Orange, California 92866, USA. E-mail: sumida@chapman.edu.

Summary: The relative impact of chronic alcohol consumption on hepatic gluconeogenesis (HGN) males and females is unknown. To determine the effects of chronic alcohol consumption (8 weeks) on HGN, the isolated liver perfusion technique was used on male and female Wistar rats fasted for 24 hours. After surgical isolation, livers were perfused (single pass) for 30 minutes with Krebs-Henseleit bicarbonate buffer and fresh bovine erythrocytes with no added substrate (washout period). After the washout period, livers were perfused with lactate (10 mM) and [U-14C]lactate (15,000 dpm/ml) using the recirculation method. Males and females fed the control diet did not differ significantly in HGN. In contrast, the females chronically fed the ethanol diet (FE) had significantly lower HGN rates (2.73 ± 0.37 μmol/min x g liver protein-1), whereas males fed the ethanol diet (ME) had significantly higher HGN rates (4.99 ± 0.45 μmol/min x g liver protein-1) than controls (3.83 ± 0.34 μmol/min x g liver protein-1). Concomitant decreases were also observed for both 14C-lactate incorporation into 14C-glucose and rates of lactate uptake for FE, while corresponding increases were observed for 14C-lactate incorporation into 14C-glucose for ME. The livers from ME were able to convert a greater percentage of the lactate into glucose, resulting in the elevation in gluconeogenic capacity. In conclusion, chronic alcohol consumption lowers the hepatic gluconeogenic capacity from lactate in females and elevates HGN in males. NIAAA Glossary Terms:   gluconeogenesis, liver function, ethanol, chronic AODE, gender differences, laboratory rat, lactate, controlled study, comparative study, glucose, metabolism, animal study

Mukamal, Kenneth J.; Massaro, Joseph M.; Ault, Kenneth A.; Mittleman, Murray A.; Sutherland, Patrice A.; Lipinska, Izabella; Levy, Daniel; D'Agostino, Ralph B.; and Tofler, Geoffrey H.  Alcohol consumption and platelet activation and aggregation among women and men: The Framingham Offspring Study.  Alcoholism: Clinical & Experimental Research 29(10):1906-1912, October 2005.

Address correspondence to Kenneth J. Mukamal, Department of Medicine, Beth Israel Deaconess Medical Center, Boston, Massachusetts 02215, USA. E-mail: kmukamal@bidmc.harvard.edu.

Summary: Alcohol intake has been associated with lower platelet activity; however, few large-scale studies have included women, and it appears that the relationship of alcohol intake with measures of platelet activation has not been studied. This study was a cross-sectional analysis of adults free of cardiovascular disease enrolled in the Framingham Offspring Study. Alcohol consumption was assessed with a standardized questionnaire. Platelet activation was measured in response to 1 and 5 μmol of adenosine diphosphate (ADP) with a P-selectin assay among 1,037 participants and platelet aggregability in response to ADP, epinephrine, and collagen among 2,013 participants. Alcohol consumption was inversely associated with P-selectin expression in response to 1 μmol ADP (p = 0.007) and 5 μmol ADP (p = 0.02) among men but not among women. Alcohol consumption was also inversely associated with platelet aggregation induced by ADP among both women (p = 0.04) and men (p for trend = 0.008) and by epinephrine among men (p = 0.03). Thus alcohol consumption is inversely associated with both platelet activation and aggregation, particularly in men. Additional research is needed to determine whether these findings contribute to the contrasting associations of alcohol consumption with risk of thrombotic and hemorrhagic cardiovascular events. NIAAA Glossary Terms:   ethanol, platelet aggregation, platelet-activating factor, gender differences, ADP, AOD consumption, epinephrine, collagen, longitudinal study, comparative study, human study

Hayashino, Yasuaki.  Is herbal "root" effective for reducing alcohol drinking?  Alcoholism: Clinical & Experimental Research 29(10):1913, October 2005.  Scott E. Lukas, Scott E. and Lee, David Y-W. Reply.  Alcoholism: Clinical & Experimental Research 29(10):1914, October 2005.

(Letters to the editor. No abstracts available.)

Zhang, Ting; Guo, Chang-Jiang; Douglas, Steven D.; Metzger, David S.; O'Brien, Charles P.; Li, Yuan; Wang, Yan-Jian; Wang, Xu; and Ho, Wen-Zhe.  Alcohol suppresses IL-2-induced CC chemokine production by natural killer cells.  Alcoholism: Clinical & Experimental Research 29(9):1559-1567, September 2005.

Summary: The authors investigated whether ethanol impairs the functioning of natural killer (NK) cells, particularly production of CC chemokines induced by interleukin (IL)-2, the natural ligands for CCR5 receptor. Primary NK cells and NK cell line (YTS) were cultured for 3 hours with or without ethanol (10 to 80 mM). Culture supernatants were then harvested and used to treat human peripheral blood monocyte-derived macrophages and a HeLa cell line, which expresses CD4, CCR5, and CXCR4 receptors (MAGI cells). CC chemokine expression by YTS and primary NK cells treated with or without alcohol was analyzed with real-time reverse transcriptase-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). [Ca2+]i and Western blot assays were used to determine calcium-mediated intracellular signaling pathway and nuclear factor κB (NF-κB) p65 expression. Human immunodeficiency virus (HIV) strains (Bal and UG024) were used to infect macrophages and MAGI cells. In addition, ADA (macrophage-tropic strain) and murine leukemia virus (MLV) envelope-pseudotyped HIV infection was carried out in macrophages. HIV infectivity was determined by HIV reverse transcriptase (RT) and β-galactosidase activity assays. Ethanol inhibited IL-2-induced CC chemokine (CCL3 and CCL4) expression by NK cells, an inhibition that was shown by functional tests to be associated with diminished anti-HIV ability of NK cells. Ethanol also reduced the ability of NK cells to respond to CCL3-mediated chemotaxis. Ethanol inhibited IL-2-induced NF-κB p65 protein expression and calcium mobilization by NK cells. Ethanol, through inhibition of IL-2-induced NF-κB p65 protein expression and intracellular calcium mobilization, suppressed NK cell production of CC chemokines. This suppression of CC chemokine production was associated with diminished anti-HIV activity of NK cells. Thus, by inhibiting NK cell-mediated innate immunity against HIV, ethanol consumption may have a cofactor role in the immunopathogenesis of HIV disease. NIAAA Glossary Terms:   natural killer cell, ethanol, cytokines, interleukin-2, cultured cell line, cell culture study, ligand, macrophage, reverse transcriptase polymerase chain reaction, immunoassay, enzymes, Western blotting, cell signaling, human immunodeficiency virus, HIV infection, chemotaxis, proteins, immune system, immune response, laboratory mice, animal study

MacLaren, Erik J. and Sikela, James M.  Cerebellar gene expression profiling and eQTL analysis in inbred mouse strains selected for ethanol sensitivity.  Alcoholism: Clinical & Experimental Research 29(9):1568-1579, September 2005.

Summary: The expression levels of over 39,000 transcripts in the cerebellum of Inbred Long-Sleep (ILS) and Inbred Short-Sleep (ISS) mice were examined to find differentially expressed (DE) candidate genes for these phenotypes. The cerebellum is a major target of ethanol's actions in the central nervous system. DE genes between the strains were identified using oligonucleotide arrays and complimentary deoxyribonucleic acid (DNA) arrays. DE genes in the mouse genome assembly were located using sequence alignment. In silico expression quantitative trait loci (eQTL) mapping was used to identify chromosomal regions likely to control the transcription level of DE genes, and overrepresented functional themes were identified by the EASE program. The genomic region immediately upstream of the cyclase associated protein homolog 1 (Cap1) gene was directly sequenced from polymerase chain reaction products. Nearly 300 genes were identified as differentially expressed in the cerebellums of ILS and ISS mice. These genes and their corresponding eQTLs map to genomic regions linked to several phenotypes that differ between the ILS and ISS strains, including ethanol preference and cocaine-induced locomotor activation on chromosomes 4 and 7. Eight genes were cross-platform validated, four of which are more highly expressed in ILS cerebellum. Three single nucleotide polymorphisms, one of which disrupts a predicted Sp1 binding site, were found in the upstream region of Cap1, a strong candidate for influencing ethanol phenotypes. Many of these DE genes are candidates for influencing ethanol- and drug-regulated phenotypes because they either map to ethanol related QTLs in the genome or are linked to them through eQTL mapping. Genes involved in calcium ion binding and transcriptional regulation are overrepresented and  therefore may influence ethanol behaviors in mice and humans. NIAAA Glossary Terms:  animal strains, laboratory mice, selective breeding, sleep, ethanol, cerebellum, central nervous system, phenotype, gene, gene expression, oligonucleotide, cDNA, gene transcription, quantitative trait loci, chromosome, genome, polymerase chain reaction, single nucleotide polymorphism, cocaine, animal selectively bred for AOD preference, locomotion, calcium, animal behavior, animal model

Radcliffe, Richard A.; Floyd, Kirsten L.; Drahnak, Joseph A.; and Deitrich, Richard A.  Genetic dissociation between ethanol sensitivity and rapid tolerance in mouse and rat strains selectively bred for differential ethanol sensitivity.  Alcoholism: Clinical & Experimental Research 29(9):1580-1589, September 2005.

Summary: A study was undertaken to determine whether acute sensitivity in Inbred Long- and Short-Sleep mice (ILS and ISS) and High- and Low-Alcohol-Senstive rats (IHAS and ILAS)  is genetically correlated to a rapid tolerance to alcohol. Separate groups of animals were administered a single pretreatment dose of alcohol (0-6 g/kg for the mice and 0-4 g/kg for the rats). Alcohol sensitivity was tested 24 hours later with the loss -of-righting-reflex (LORR) test, and blood ethanol concentration was tested at regain of righting (BECRR). Ethanol-induced hypothermia also was determined in the mice. Independently derived replicate rat strains were used for all experiments (IHAS1, ILAS1; IHAS2, ILAS2) (such replicates do not exist for ILS and ISS mice).  Alcohol pretreatment dose-dependently decreased LORR duration and increased BECRR in the IHAS1 and ILS strain, suggesting the development of functional rapid tolerance. In contrast, LORR duration increased in the ILAS1, ILAS2, and ISS groups, but BECRR either increased (ILAS1, ILAS2) or did not change (ISS). These observations suggest that central nervous system sensitivity was decreased in the ILAS1 and ILAS2 groups (i.e., rapid functional tolerance) or unchanged in the ISS strain, but that some pharmacokinetic property also was altered in these strains. The results do not support a genetic relation between alcohol sensitivity and the development of rapid tolerance. NIAAA Glossary Terms:  animal strains, selective breeding, laboratory mice, laboratory rat, sleep, ethanol, AOD sensitivity, AOD tolerance, genetic correlation analysis, righting reflex, BAC, hypothermia, central nervous system, pharmacokinetics, animal model

De Bellis, Michael D.; Narasimhan, Anandhi; Thatcher, Dawn L.; Keshavan, Matcheri S.; Soloff, Paul; and Clark, Duncan B.  Prefrontal cortex, thalamus, and cerebellar volumes in adolescents and young adults with adolescent-onset alcohol use disorders and comorbid mental disorders.  Alcoholism: Clinical & Experimental Research 29(9):1590-1600, September 2005.

Summary: Prefrontal, thalamic, and cerebellar brain injury is associated with excessive ethanol intake in adults. This study tested the hypothesis that subjects with adolescent-onset alcohol use disorders (AUD) would have smaller volumes in these brain areas than sociodemographically similar control subjects. Magnetic resonance imaging was used to measure prefrontal cortex, thalamic, and cerebellar volumes in subjects with an AUD (n = 14; 8 males and 6 females; mean age, 17.0 ± 2.1 years) and 28 control subjects (n = 28; 16 males and 12 females; 16.9 ± 2.3 years). All AUD subjects were recruited from substance abuse treatment programs and had comorbid mental disorders. Subjects with an AUD had smaller prefrontal cortex and prefrontal cortex white matter volumes compared with controls. The groups did not differ in right, left, and total thalamic, pons/brainstem, right and left cerebellar hemispheric, total cerebellar, and cerebellar vermis volumes. There was a significant sex-by-group effect, indicating that males with an adolescent-onset AUD compared with control males had smaller cerebellar volumes, whereas the two female groups did not differ in cerebellar volumes. Prefrontal cortex volume variables correlated significantly with measures of alcohol consumption. These findings suggest that a smaller prefrontal cortex is associated with early-onset drinking in individuals with comorbid mental disorders. The authors recommend further studies to examine whether a smaller prefrontal cortex volume represents a vulnerability to early-onset drinking or is a consequence of it. NIAAA Glossary Terms:  prefrontal cortex, thalamus, cerebellum, central nervous system, brain damage, hazardous drinking, hypothesis testing, adolescence, early AODU onset, underage drinking, alcohol use disorder classification, brain atrophy, magnetic resonance imaging, brain imaging, treatment program, case-control study, comorbidity, mentally ill, pons, brainstem, gender differences, correlation analysis, human study

Sharpe, Amanda L.; Coste, Sarah C.; Burkhart-Kasch, Sue; Li, Na; Stenzel-Poore, Mary P.; and Phillips, Tamara J.  Mice deficient in corticotropin-releasing factor receptor type 2 exhibit normal ethanol-associated behaviors.  Alcoholism: Clinical & Experimental Research 29(9):1601-1609, September 2005.

Summary: Stress is believed to influence alcohol use and relapse in alcoholics. Animal studies suggest an interaction between corticotropin-releasing factor (CRF) and its receptors and the behavioral effects and consumption of alcohol. This study examined the effect of CRF receptor type 2 (CRF2) on ethanol consumption, conditioned taste aversion, sedation, and hypothermia. CRF2-null mutant or knock-out (KO), and wild-type (WT) mice were used to assess consumption of increasing concentrations of ethanol in a two-bottle, 24-hour test and during daily limited-access sessions. Ethanol-induced conditioned taste aversion (CTA), loss of righting reflex (LORR), hypothermia, and ethanol metabolism kinetics were also examined in the two groups. CRF2 KO mice did not differ from WT mice in sensitivity to ethanol-induced CTA, LORR, hypothermia, or ethanol metabolism kinetics. There was no genotypic difference in ethanol intake or preference in the 24-hour, two-bottle choice procedure, and only modestly reduced consumption of the 7.5 and 10% ethanol solutions in KO versus WT mice in the limited-access procedure. The observation that CRF2 deficiency had little effect on several ethanol-associated behaviors in CRF2 KO mice suggests that this receptor does not have a primary role in modulating these behaviors. Evidence of a role for this receptor in neural circuits subserving stress-coping behaviors suggest that future studies should focus on the role of endogenous CRF2 in ethanol-associated behaviors in mice that are stressed or withdrawing from dependence on ethanol. NIAAA Glossary Terms:  AOD use, AODD relapse, AOD dependence, corticotropin RH, hormone receptors, behavior, AOD consumption, ethanol, aversion conditioning, taste conditioning, sedative-hypnotics, hypothermia, knockout gene technology, righting reflex, ethanol metabolish, pharmacokinetics, AOD sensitivity, genotype, AOD preference, laboratory mice, animal study

Weitemier, Adam Z. and Ryabinin, Andrey E.  Brain region-specific regulation of urocortin 1 innervation and corticotropin-releasing factor receptor type 2 binding by ethanol exposure.  Alcoholism: Clinical & Experimental Research 29(9):1610-1620, September 2005.

Summary: Ethanol administration and consumption selectively activates the urocortin 1 (Ucn1)-expressing neurons of the Edinger-Westphal nucleus. The authors of this study investigated whether repeated ethanol exposure affects Ucn1 and Ucn1-responsive corticotropin-releasing factor type-2 receptors (CRF2). Male C57BL/6J and DBA/2J mice were injected intraperitoneally with ethanol (2 g/kg) once a day for 14, 7, or 0 days. Ucn1 immunoreactivity was measured in the lateral septum, dorsal raphe, and Edinger-Westphal nucleus. In a separate experiment, C57BL/6J mice were exposed to ethanol for 7, 1, or 0 days, and CRF2 receptor binding was measured in the lateral septum and dorsal raphe by receptor autoradiography. Ethanol administration induced parallel changes in Ucn1 immunoreactive terminal fibers in the lateral septum and dorsal raphe of both strains. Seven ethanol exposures but not one ethanol exposure significantly increased CRF2 receptor binding in the dorsal raphe and slightly increased CRF2 receptor binding in the lateral septum. The results provide evidence that the Ucn1/CRF2 receptor system can be modified by ethanol and suggest that this system may be involved in behavioral changes during alcoholism. NIAAA Glossary Terms:  ethanol, AOD consumption, neuron, brain, corticotropin RH, hormone receptors, intraperitoneal administration, laboratory mice, immune response, radiography, AOD use behavior, AOD dependence, animal study

Ginsburg, Brett C. and Lamb, R J.  Alphaxalone and epiallopregnanolone in rats trained to discriminate ethanol.  Alcoholism: Clinical & Experimental Research 29(9):1621-1629, September 2005.

Summary: Neurosteroids with a 3α-hydroxy orientation share pharmacological effects with ethanol, increase in brain after ethanol administration, and may mediate ethanol effects. 3β-hydroxy neurosteroids antagonize in vitro and some in vivo effects of ethanol and 3α-hydroxy neurosteroids. This study assessed the discriminative stimulus and rate-altering effects of alphaxalone, a 3α-hydroxy neurosteroid, and epiallopregnanolone, a 3β-hydroxy neurosteroid, in rats trained to discriminate either 0.8 g/kg or 1.2 g/kg of ethanol. The ability of epiallopregnanolone to antagonize the discriminative stimulus or rate-altering effects of ethanol or alphaxalone was also assessed. Ethanol had a similar discriminative ED50 (0.5 g/kg) in both groups, but rats trained with the lower ethanol dose were more sensitive to ethanol's rate-decreasing effects. Alphaxalone produced ethanol-appropriate responding in both training groups, although less effectively in rats trained on the lower ethanol dose (maximum 65% versus 80% ethanol-appropriate responding). The groups did not differ in sensitivity to the rate-decreasing effects of alphaxalone. Epiallopregnanolone did not reliably produce ethanol-appropriate responding in either training group, and rats trained on the lower ethanol dose were slightly more sensitive to epiallopregnanolone's rate-decreasing effects. Epiallopregnanolone did not alter any effects of ethanol or alphaxalone. The results agree with previous reports that 3α-hydroxy neurosteroids produce ethanol-appropriate responding, while 3β-hydroxy neurosteroids do not; as well as reports showing no antagonism of the discriminative stimulus or rate-suppressant effects of ethanol or 3α-hydroxy neurosteroids by 3β-hydroxy neurosteroids. Although the results demonstrate that ethanol and 3α-hydroxy neurosteroids share discriminative stimulus effects, they are inconsistent with the hypothesis that such neurosteroids mediate ethanol's discriminative stimulus. NIAAA Glossary Terms:   neurosteroids, steroid hormones, ethanol, drug discrimination, CNS stimulants, antagonists, AOD sensitivity, dose-response relationship, hypothesis testing, laboratory rat, animal study

Ford, Matthew M.; Nickel, Jeffrey D.; Phillips, Tamara J.; and Finn, Deborah A.  Neurosteroid modulators of GABA_A receptors differentially modulate ethanol intake patterns in male C57BL/6J mice.  Alcoholism: Clinical & Experimental Research 29(9):1630-1640, September 2005.

Summary: Allopregnanolone (ALLO) and structurally-related endogenous neurosteroids are potent modulators of gamma-aminobutyric acid A (GABAA) receptor function at physiologically relevant concentrations. Growing evidence implicates a modulatory role for ALLO in behavioral processes underlying ethanol self-administration, discrimination, and reinstatement. This study evaluated the impact of exogenous neurosteroid challenges with the agonist ALLO and the partial agonist/antagonist epipregnanolone (EPI) on ethanol drinking patterns. Male C57BL/6J mice were initiated to consume an unsweetened 10% v/v ethanol solution (10E) by a saccharin fading procedure during daily 2-hour limited-access sessions beginning 1-hour after dark-phase onset. Cumulative lick responses were recorded for 10E and water by lickometer circuits. After establishing 10E intake baselines, mice were habituated to intraperitoneal vehicle injection (VEH; 20% w/v β-cyclodextrin), then were treated with either VEH or neurosteroid immediately before the drinking session. Each mouse received a series of ALLO doses (3.2, 10, 17, and 24 mg/kg) alone and EPI doses (0.15, 1, 3, and 10 mg/kg) alone in a counterbalanced within-group design. The GABAA receptor-positive modulator ALLO dose-dependently modulated overall ethanol intake throughout the 2-hour session with the 3.2 mg/kg dose eliciting a significant increase, whereas the 24 mg/kg dose produced a significant suppression of ethanol intake versus VEH pretreatment. ALLO-evoked intake alterations corresponded to significant, dose-dependent alterations in bout frequency and interbout interval. ALLO also elicited robust, dose-dependent elevations in 10E licks during the initial 5 minutes of access but subsequently produced in a dose-dependent suppression of 10E licks during session minutes 20-80. In contrast, the partial agonist/antagonist neurosteroid EPI exhibited no influence on any consumption parameter evaluated. The results suggest that GABAA receptor-active neurosteroids may modulate the regulatory processes that govern the onset, maintenance, and termination of drinking episodes. The differential influence of ALLO and EPI on ethanol intake patterns may reflect an alteration in GABAergic inhibitory tone that is likely due to each neurosteroid's pharmacological profile at GABAA receptors. Manipulation of endogenous ALLO may prove useful for reducing excessive intake and protecting against the loss of regulatory control over drinking. NIAAA Glossary Terms:   GABA A receptor, GABAergic neuron, allopregnanolone, ethanol, neurosteroids, steroid hormones, animal behavior, laboratory mice, intraperitoneal administration, agonists, antagonists, AOD consumption, AOD intake per occasion, dose-response relationship, animal study

Brunell, Steven C. and Spear, Linda P.  Effect of stress on the voluntary intake of a sweetened ethanol solution in pair-housed adolescent and adult rats.  Alcoholism: Clinical & Experimental Research 29(9):1641-1653, September 2005.

Summary: Previous studies reporting greater voluntary ethanol intake in adolescent than adult rats have examined intake in isolate-housed animals. Because the stress of isolate housing may differ ontogenetically as well as confound interpretation of other stressor effects, this study examined stressor/ethanol interactions among pair-housed adolescent and adult rats. Sprague-Dawley male rats were implanted with identification tags to allow individual monitoring of homecage intake of water and either a 10% (v/v) ethanol solution containing 0.1% (w/v) saccharin or saccharin alone over a 14-day access period. Animals were given 0, 1, or 8 daily 15-minute footshock sessions, with shock-induced freezing and pre-, post-, and recovery corticosterone levels determined on the first and last footshock exposure days. After the access period, withdrawal was assessed with a plus maze, and tolerance to ethanol-induced loss of righting reflex was examined. Nonstressed adolescents drank considerably more sweetened ethanol than did adults, with chronic stress suppressing this adolescent consumption. Ethanol access in adolescents disrupted within-session adaptation to footshock in terms of freezing behavior, although no such disruption was evident at either age when indexed hormonally. Despite relatively high ethanol intakes (up to 6 g/kg/day in the adolescents), there was no evidence of withdrawal-associated anxiogenesis. Evidence of tolerance was mixed and when present was metabolic in nature. It was concluded that previous reports of heightened voluntary ethanol intake among adolescent rats are not a function of isolate stress. Adolescents were more sensitive than adults to ethanol/stress interactions, with the elevated ethanol intake of pair-housed adolescents selectively disrupted by chronic stress, a disruption not evident in adults. Likewise, ethanol disrupted behavioral adaptation to the footshock stressor among adolescents but not adults. NIAAA Glossary Terms:  laboratory rat, age differences, adolescent, adult, ethanol, stress, stressor, AOD intake per occasion, social isolation, confounding variable, saccharin, anxiety, animal study

Martin-Garcia, Elena and Pallares, Marc.  Effects of intrahippocampal nicotine and neurosteroid administration on withdrawal in voluntary and chronic alcohol-drinking rats.  Alcoholism: Clinical & Experimental Research 29(9):1654-1663, September 2005.

Summary: It has been shown in previous studies that 4.6 μg of nicotine administered to the hippocampus can deteriorate learning acquisition in alcohol-drinking rats. This study investigated whether this nicotine dose can alter the alcohol withdrawal syndrome and whether these effects can be modulated by the neurosteroids allopregnanolone (AlloP) and pregnenolone sulfate (PregS), at doses previously reported as anxiolytic and promnesic respectively. A free-choice drinking procedure was used that involved providing the rats with an alcoholic solution (10% ethanol) at an early age. Alcohol and control rats were assigned randomly to six groups that received two consecutive intrahippocampal (dorsal CA1) injections once a week during three consecutive weeks after 1 hour of ethanol drinking. The first injection was nicotine (4.6 μg, 20 mM) or saline and the second injection was PregS (5 ng, 24 μM), AlloP (0.2 μg, 1.26 μM) or saline. Blood alcohol concentrations were assessed one week before the withdrawal testing. Locomotor activity and audiogenic seizures were tested during withdrawal after 110 days of voluntary ethanol consumption. Rats were injected immediately before the withdrawal testing. AlloP induced a decrease in horizontal and vertical activities, suggesting that the dose tested has sedative effects. AlloP reversed the seizures induced by ethanol withdrawal and also the spontaneous audiogenic seizures induced by the acoustic stimulation in control rats. Moreover, AlloP decreased other alcohol withdrawal signs, such as tail stiffening and body rigidity. Intrahippocampal administration of nicotine or PregS, at the doses tested, did not effectively modify the expression of audiogenic seizures induced by alcohol withdrawal. The results show that hippocampal GABAergic activity and AlloP have an important role in preventing convulsive behavior. The results also highlight the therapeutic potential of AlloP for reducing the alcohol withdrawal syndrome. NIAAA Glossary Terms:  nicotine, hippocampus, AOD withdrawal syndrome, laboratory rat, neurosteroids, steroid hormones, allopregnanolone, pregnenolone, BAC, locomotion, AODR seizure, sound, sedative-hypnotics, convulsion, animal study

Ilgen, Mark A.; Tiet, Quyen; Finney, John W.; and Harris, Alex H. S.