Alcohol drinking in rats injected ICV with 6-OHDA: Effect of 8-OHDPAT and tropisetron (ICS 205930)

6-Hydroxydopamine (6-OHDA) was administered ICV to Wistar male rats. Lesioned animals displayed lower preference for ethanol (ETOH) than sham-operated rats. Among 6-OHDA lesioned rats only 9% became high-preferring whereas 20% of sham-operated animals became high-preferring ones. Both tropisetron (the antagonist of 5-HT₃ receptors) and 8-OHDPAT (the 5-HT_1A receptor agonist) reduced ETOH drinking in high-preferring rats. However, in 6-OHDA lesioned rats the effect of tropisetron was reduced although 8-OHDA retained its effect on ETOH consumption. These results suggest that brain DA neurons are involved in tropisetron action but are not responsible for antipreference effect of 8-OHDPAT.     

Adaptive changes of dopamine-D2 receptors in rat brain following ethanol withdrawal: a quantitative autoradiographic investigation.

The effect of subchronic treatment with two doses of ethanol (5 and 10 vol% drinking fluid) on the density of dopamine-D2 receptors was investigated at two different phases of withdrawal, namely 24 h and 5 days after the cessation of the ethanol application. The number of dopamine-D2 receptors was affected in regions receiving projections from both the substantia nigra as well as the ventral tegmentum. Twenty-four hours after the replacement of the ethanol solution by water, a dose-dependent decrease of D2 receptors was found in all regions (N. caudatus dorsalis, medialis and ventralis, N. accumbens lateralis and medialis, tuberculum olfactorium) and most of the analyzed planes [interaural 7.7-10.2 according to the atlas of Paxinos and Watson (35)]. At day 5 of withdrawal, the number of dopamine-D2 receptors of the animals treated with 5 vol% ethanol reached the level of water controls in most planes. In contrast, two- to three-fold higher numbers were detected in animals treated with the higher dose. Only in the most caudal parts of the investigated regions, was the number of receptors decreased with the higher dose. The mesocorticolimbic system seems to be less sensitive to the effects of ethanol than the nigrostriatal neurones. The findings of the present study suggest an increased activity of dopaminergic neurons with an adaptive reduction of dopamine-D2 receptors during the subchronic treatment with ethanol during the first day(s) of withdrawal. This phase is followed by a reduced turnover rate for up to 7 days (21). The reduced activity of dopaminergic neurones induces a compensatory increase of the number of receptors.(ABSTRACT TRUNCATED AT 250 WORDS)     

Female Syrian golden hamster: drinking of high concentrations of ethanol aversive to other mammals

The present experiments were designed to determine: 1) the pattern of preference for different concentrations of ethanol in the female Syrian golden hamster (Mesocricetus auratus), and 2) the influence of drinking ethanol on their intakes of food and total calories. A standard three-bottle preference test was undertaken in six female hamsters over an 11-day period in which water was offered together with ethanol, which was increased in concentration over 11 clays from 3% to 50% as follows: 3%, 5%, 7%, 9%, 12%, 15%, 20%, 25%, 30%, 40%, 50%. Then, each hamster was offered its individually preferred concentration for a period of 8 days until the intake of ethanol had stabilized. During the preference testing for 3–25% solutions, the proportional intakes ranged between 0.6 and 0.8 whereas the mean absolute amount consumed per day increased from 2.3 to 16.1 g/kg at the 25% concentration. However, at the 50% concentration, ethanol drinking declined substantially to 8.7 g/kg per day. The overall mean percent concentration of ethanol preferred by the hamsters was 24.2 ± 1.5%. During the following 8-day period when the maximally preferred concentration of ethanol of each hamster was offered with water, the mean intake of ethanol was 17.9 ± 1.1 g/kg per day. Throughout the test sequence, the caloric intake of the animals was maintained in that calories obtained from food declined at the same rate as the calories obtained from ethanol in rising concentrations. These results reveal that the preference for ethanol of the female hamster is well above that reported in earlier studies in which preference was defined in terms of the proportion of ethanol to total fluid intake. Differences in procedure could account for these findings because the present experiments utilized single-animal housing, a triple bottle method, and measurement of g/kg per day obtained at 11 different concentrations of ethanol.     

Sex differences in pattern of drinking.

Drinking patterns of male and female Long-Evans rats were compared during a 15-day drinking period. All animals were tested for preference for alcohol for 24 h during which food, water, and beer containing 5% ethanol were freely available. Animals drinking 50 ml or more of beer were chosen for the experiments. On days 1-5, animals were offered food, water, and beer containing 5% ethanol (v/v). On days 6-15, the concentration of ethanol in the beer was doubled to 10% (v/v). Preference ratios (beer/total fluid) were higher for females than males, and females consumed more grams of alcohol per unit of body weight. When alcohol concentration was doubled, females increased alcohol intake (g/kg), while males tended to titrate alcohol intake to levels consumed at 5% concentration. Female patterns of drinking differed from male patterns of drinking.     

Short-term supplementation with plant extracts rich in flavonoids protect nigrostriatal dopaminergic neurons in a rat model of Parkinson's disease

OBJECTIVE: Antioxidants from plants were known to reduce the oxidative stress by scavenging free radicals, chelating metal ions and reducing inflammation. As increased oxidative stress was implicated in the nigrostriatal dopaminergic neuronal loss in Parkinson's disease (PD), we have assessed whether the plant extracts protects the nigrostriatal dopaminergic neurons in the animal model of PD. METHODS: Male adult Sprague-Dawley rats were treated orally between 10 am-11 am each day with the extracts from tangerine peel, grape seeds, cocoa and red clover for four days. One hour after the final dosing, the left medial forebrain bundle was lesioned by infusing the dopaminergic neurotoxin 6-hydroxydopamine (6-OHDA; 12 microg) under anaesthesia. Seven days post-lesion, the number of dopaminergic cells in the substantia nigra pars compacta and the levels of dopamine and its metabolites 3, 4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the striata were quantified and compared with the vehicle-treated groups. RESULTS: Compared to the unlesioned side, 6-OHDA lesions significantly reduced the number of dopaminergic cells and the levels of dopamine and its metabolites DOPAC and HVA in the vehicle-treated animals. Pretreatment of animals with extracts of tangerine peel (rich in polymethoxylated flavones; 35 mg/kg/day), cocoa-2 (rich in procyanidins; 100 mg/kg/day) and red clover (rich in isoflavones; 200 mg/kg/day) significantly attenuated the 6-OHDA-induced dopaminergic loss. However, no significant protection was seen in animals supplemented with red and white grape seeds (rich in catechins; 100 mg/kg/day), and cocoa-1 (rich in catechins; 100 mg/kg/day). CONCLUSIONS: Pre-treatment of plant extracts rich in polymethoxylated flavones, procyanidins and isoflavones but not catechins protected the nigrostriatal dopaminergic neurons in the rat model of PD.     

Age dependent development of ethanol drinking in rats after inhibition of aldehyde dehydrogenase.

The purpose of this experiment was to determine the temporal characteristics associated with the age-related development of volitional consumption of ethanol induced by the pharmacological inhibition of aldehyde dehydrogenase (AlDH). To induce preference for ethanol, the AlDH inhibitor, cyanamide, was administered to male Sprague-Dawley rats which were 30 days of age. Cyanamide (n = 8) was injected subcutaneously twice daily in a dose of 10 mg/kg over a period of 3 days while the control group (n = 6) received the saline vehicle solution according to the same schedule. Then at 50, 70, 90, and 110 days of age, both groups of rats were given a standard 11-day test of preference for water versus ethanol offered in concentrations ranging from 3% through 30%. The results showed that at 70 days of age the preference for ethanol increased above the level of the 50-day test in terms of absolute g/kg intakes and proportion of ethanol to water consumed over the lower range of 3% through 15% concentrations. During the tests at 90 and 110 days of age, the cyanamide-treated rats further increased their preference for ethanol significantly over the levels at the 70-day test in terms of both g/kg and proportional intakes. The pattern of drinking of ethanol offered in the higher concentrations of 25% and 30% was unrelated to the age of the rats and the overall intakes were significantly higher than those of the lower concentrations. These findings demonstrate that the enzymatic inhibition of AlDH systematically acts in a delayed fashion to shift the pattern of preference for ethanol which is contingent on the maturation of the animal. In this instance, the volitional intake of ethanol in the cyanamide-treated rats reached its maximal level by 90-110 days of age. It is proposed that an endocrine mechanism involved in gonadal maturation may function in the intense shift in alcohol drinking.     

Characterization of the ethanol-deprivation effect in substrains of C57BL/6 mice.

Ethanol craving plays a major role in relapse drinking behavior. Relapse and ethanol craving are an important focus for the treatment of alcoholism. The ethanol-deprivation effect (EDE) is a widely used animal model of alcohol craving. While the EDE is widely studied in rats, the molecular mechanisms underlying EDE are not clearly understood. The C57BL/6 inbred mouse strain is widely used for behavioral and molecular analyses of ethanol drinking but studies on the EDE have not been reported in this strain. In the present study, we characterized a simple behavioral protocol that rapidly and reliably induced EDE in C57BL/6 mice. Briefly, single-housed adult male C57BL/6NCrl and C57BL/6J mice were presented at the beginning of dark phase with two-bottle choice drinking containing either 10% wt/vol ethanol or tap water for 18 h/day, as well as food ad libitum. Following ethanol drinking for 4 days or 14 days, mice were deprived of ethanol for a period of 4 days. To study EDE, mice were reinstated with two bottles containing either ethanol (10% wt/vol) or water. Mice were exposed to single or multiple ethanol-deprivation cycles. Ethanol consumption (g/kg/18 h) and percent ethanol preference (% preference/18 hrs) was recorded for individual mice. C57BL/6NCrl mice consumed moderate amounts (4.78+/-0.63 g/kg) of ethanol but showed robust EDE after ethanol-drinking episodes (4 days or 14 days) as evidenced by increased ethanol consumption and ethanol preference following reinstatement of ethanol. While repeated ethanol deprivation in C57BL/6NCrl mice transiently increased ethanol consumption and ethanol preference, the magnitude of these behaviors was reduced as compared to the first deprivation cycle. In contrast, the C57BL/6J substrain consumed substantially higher levels (9.65+/-0.90 g/kg) of ethanol but did not show a clear EDE after single or multiple ethanol-deprivation cycles. In conclusion, we established a simple and reliable behavioral model to study EDE in C57BL/6NCrl mice. A reliable behavioral model to study EDE in inbred C57BL/6NCrl mice could greatly facilitate further studies on molecular mechanisms of ethanol craving behavior.     

Drinking of high concentrations of ethanol versus palatable fluids in alcohol-preferring (P) rats: valid animal model of alcoholism

A genetically based animal model of alcoholism has been characterized in Wistar-derived rats in terms of their preference (P rats) or lack of preference (NP rats) for 10% ethanol over water. The present experiments were designed to determine: 1) whether a 10% solution of ethanol is the optimal concentration for differentiation of these lines; 2) what concentrations of ethanol are maximally preferred by P and NP rats; and 3) whether highly palatable fluids presented simultaneously with each rat's preferred solution of ethanol would alter the patterns of drinking by either the P or NP or both lines of rats. A three-bottle procedure was used to establish preference for ethanol in the presence of water as well as highly palatable solutions. The results showed that, when concentrations ranging from 3-30% were presented over a 12-day test interval, the mean absolute intake of ethanol of the P rats was 6.7 g/kg per day, with a maximum intake of 10.9 g/kg per day at the 25% concentration. These levels of intake were significantly higher than the 4.3 g/kg per day consumed during the presentation of the commonly used constant concentration of 10%. Similarly, the mean absolute intake of ethanol by the NP rats was also elevated significantly at concentrations of 15-30% (2.0 g/kg per day) above that consumed at the 10% concentration (0.4 g/kg).(ABSTRACT TRUNCATED AT 250 WORDS)     

Irreversible suppression of alcohol drinking in cyanamide-treated rats after sustained delivery of the 5-HT2 antagonist amperozide

The purpose of this study was to evaluate the long-term effect of sustained treatment with amperozide, which has been shown to attenuate the volitional drinking of ethyl alcohol in the rat without side effects. Preference for alcohol first was induced pharmacologically in Sprague-Dawley rats by the inhibitor of aldehyde dehydrogenase, cyanamide, administered in a dose of 10 mg/kg twice daily for 3 days. Then following a standard preference test, each rat was offered water and its maximally preferred concentration of alcohol which ranged from 7% to 15%. Following a 4-day pre-drug test, saline control vehicle or amperozide was administered for 7 days by an osmotic minipump implanted in the intrascapular space. A single dose of 208 μg/kg/h (i.e., 5.0 mg/kg/day) was selected on the basis of a prior dose response study of amperozide. During the interval of sustained release of amperozide, the consumption of alcohol declined significantly in terms of both absolute g/kg intake and proportion of alcohol to water. When the preference of the rats was retested at 4, 30, 70, 110, and 140 day intervals after the pump had exhausted amperozide, the absolute g/kg consumption of alcohol continued to decline significantly. Unlike other drugs, amperozide did not produce any side effects, particularly on the intake of food or water or on body weight, which suggests a pharmacological specificity of its action. Because amperozide acts centrally on 5-HT₂ receptors as well as on dopaminergic synapses in the limbic system, it is envisaged that the drug exerts a unique effect on reward systems in the brain by affecting their receptor reuptake mechanisms, release of the respective transmitters, or other processes potentially involved in the abnormal imbibition of alcohol. Finally, because the effect of amperozide on alcohol drinking is progressive and irreversible, it thus may serve as a pharmacological adjunct to current therapy used in the clinical treatment of the disease of alcoholism.     

Alcohol preference drinking in a mouse line selectively bred for high drinking in the dark

We have selectively bred mice that reach very high blood ethanol concentrations (BECs) after drinking from a single bottle of 20% ethanol. High Drinking in the Dark (HDID-1) mice drink nearly 6 g/kg ethanol in 4 h and reach average BECs of more than 1.0 mg/mL. Previous studies suggest that DID and two-bottle preference for 10% ethanol with continuous access are influenced by many of the same genes. We therefore asked whether HDID-1 mice would differ from the HS/Npt control stock on two-bottle preference drinking. We serially offered mice access to 3-40% ethanol in tap water versus tap water. For ethanol concentrations between 3 and 20%, HDID-1 and HS/Npt controls did not differ in two-bottle preference drinking. At the highest concentrations, the HS/Npt mice drank more than the HDID-1 mice. We also tested the same mice for preference for two concentrations each of quinine, sucrose, and saccharin. Curiously, the mice showed preference ratios (volume of tastant/total fluid drunk) of about 50% for all tastants and concentrations. Thus, neither genotype showed either preference or avoidance for any tastant after high ethanol concentrations. Therefore, we compared naive groups of HDID-1 and HS/Npt mice for tastant preference. Results from this test showed that ethanol-naive mice preferred sweet fluids and avoided quinine but the genotypes did not differ. Finally, we tested HDID-1 and HS mice for an extended period for preference for 15% ethanol versus water during a 2-h access period in the dark. After several weeks, HDID-1 mice consumed significantly more than HS. We conclude that drinking in the dark shows some genetic overlap with other tests of preference drinking, but that the degree of genetic commonality depends on the model used.     

Chronic ethanol exposure leads to divergent control of dopaminergic synapses in distinct target regions

Neuroadaptations following chronic exposure to alcohol are hypothesized to play important roles in alcohol-induced alterations in behavior, in particular increased alcohol drinking and anxiety like behavior. Dopaminergic signaling plays a key role in reward-related behavior, with evidence suggesting it undergoes modification following exposure to drugs of abuse. A large literature indicates an involvement of dopaminergic signaling in response to alcohol. Using a chronic inhalation model of ethanol exposure in mice, we have begun to investigate the effects of alcohol intake on dopaminergic signaling by examining protein levels of tyrosine hydroxylase and the dopamine transporter, as well as monoamine metabolites in three different target fields of three different dopaminergic nuclei. We have focused on the dorsal lateral bed nucleus of the stria terminalis because of the reported involvement of dorsal lateral bed nucleus of the stria terminalis dopamine in ethanol intake, and the nucleus accumbens and dorsal striatum because of their dense dopaminergic innervation. After either a chronic intermittent exposure or continuous exposure regimen, mice were killed, and tissue punches collected from the dorsal lateral bed nucleus of the stria terminalis, nucleus accumbens, and striatum for Western analysis. Strikingly, we found divergent regulation of tyrosine hydroxylase and dopamine transporter protein levels across these three regions that was dependent upon the means of exposure. These data thus suggest that distinct populations of catecholamine neurons may be differentially regulated by ethanol, and that ethanol and withdrawal interact to produce differential adaptations in these systems.     

Accumbal dopamine concentration during operant self-administration of a sucrose or a novel sucrose with ethanol solution.

The goal of the current study was to determine the effect of operant self-administration of (1) 10% sucrose and (2) a first-time solution of 10% sucrose with 5% or 10% ethanol, on dopamine concentration in the nucleus accumbens. We used an operant procedure that distinguished lever pressing (an appetitive behavior) from drinking to better assess the effect of fluid consumption on accumbal dopamine activity. Male Long-Evans rats were trained to bar press by using 10% sucrose reinforcement, and they were required to emit an escalating number of bar presses across daily sessions. Completion of the response requirement resulted in 20 min of access to the solution. Microdialysis samples were collected before, during, and after bar pressing and drinking, and content of ethanol and dopamine was determined. Dopamine concentration in the dialysate was slightly, but significantly, increased in both groups during lever pressing. However, after consumption began, dopamine concentration increased in the sucrose, but not in the sucrose with ethanol, group, followed by a return to baseline values. Ethanol consumption was low (0.27 +/- 0.02 g/kg) and corresponded to low dialysate ethanol concentrations, which appeared within 5 min of drinking. These results demonstrate that operant self-administration of sucrose increases accumbal dopamine concentration during consummatory phases of behavior, but that a similar increase is not apparent when a novel, perhaps aversive, solution (sucrose with ethanol) is presented. This difference may be due to the sensory-related stimulus properties of each solution. In addition, oral self-administration of ethanol at 0.27 +/- 0.02 g/kg over 20 min is not sufficient for stimulation of dopamine activity in the nucleus accumbens.     

Extracellular levels of dopamine in the nucleus accumbens in AA and ANA rats after reverse microdialysis of ethanol into the nucleus accumbens or ventral tegmental area.

Ethanol is known to increase the release of dopamine in the nucleus accumbens. The question of whether this is a result of a direct or an indirect effect of ethanol on mesolimbic dopaminergic neurons was examined by investigating the extracellular levels of dopamine and its metabolites in the nucleus accumbens of alcohol-preferring AA (Alko Alcohol) and alcohol-avoiding ANA (Alko Non-Alcohol) rats after application of ethanol locally into either the nucleus accumbens or the ventral tegmental area with the use of reverse microdialysis. Application of ethanol (200, 400, or 800 mM in dialysate) into the nucleus accumbens, but not into the ventral tegmental area, temporarily increased the accumbal levels of dopamine in a dose-dependent manner. The ethanol-evoked increase in the level of extracellular dopamine was more prominent in AA rats than in ANA rats. Ethanol tended to suppress levels of 3,4-dihydroxyphenylacetic acid and homovanillic acid. Because the concentrations of ethanol found to elevate the extracellular level of dopamine can hardly be considered pharmacologically relevant, the increase in accumbal dopamine levels after application of ethanol may be due to nonspecific membrane effects of ethanol. The findings support the suggestion that the increase in the extracellular level of dopamine in the nucleus accumbens after systemic administration of ethanol may involve other sites on dopamine neurons or even different neurotransmitter systems, rather than the action of ethanol at the mesolimbic dopaminergic terminals.     

Drinking Patterns in Genetic Low-Alcohol-Drinking (LAD) Rats After Systemic Cyanamide and Cerebral Injections of THP or 6-OHDA

A key question related to the role of acetaldehyde and aldehyde adducts in alcoholism concerns their relationship to the genetic mechanisms underlying drinking. Experimentally, the low-alcohol-drinking (LAD) rat represents a standard rodent model having a strong aversion to alcohol. In these experiments, preferences for water vs. alcohol, offered in concentrations from 3% to 30%, were determined over 10 days in adult LAD rats (N = 6 per group). Then a saline vehicle or either 10 or 20 mg/kg of the aldehyde dehydrogenase (AlDH) inhibitor, cyanamide, was injected SC twice daily for 3 days. Secondly, either 0.5 or 1.0 μg of tetrahydropapaveroline (THP) was infused ICV twice daily for 3 days in LAD rats (N = 8) and, as a genetic control, THP also was infused identically in Sprague–Dawley (SD) rats (N = 8). The results showed that the lower and higher doses of cyanamide augmented alcohol intakes in 33% and 50% of the LAD rats, respectively, with the patterns of drinking resembling that of genetic high-alcohol-drinking HAD or P rats. Although ICV infusions of THP had little effect on alcohol preference of LAD rats, alcohol drinking was enhanced significantly in the SD rats. In a supplementary study, 200 μg of 6-hydroxydopamine (6-OHDA) also was infused ICV in LAD rats (N = 7) on two consecutive days; no change occurred in the characteristic aversion to alcohol. These findings suggest that in certain individuals, a perturbation in the synthesis of AlDH can modify the genetically based aversion to alcohol, thus precipitating the liability for alcoholism. In that neither THP nor 6-OHDA lesioning exerted any effect on the genetic nondrinking LAD animal suggests that an unknown endogenous factor in the brain must underlie the cyanamide-induced shift to alcohol preference. We conclude that the genetic elements that normally prevent the progression to addictive drinking in most individuals appear to be invariant and irreversible.     

Brain region specific modulation of ethanol-induced depression of GABAergic neurons in the brain reward system by the nicotine receptor antagonist mecamylamine

The mechanisms underlying ethanol-induced activation of the mesolimbic dopamine system are not fully understood, but increased extracellular dopamine in the nucleus accumbens (nAc) has been shown to involve nicotinic acetylcholine receptors (nAChRs). Basal activity of dopaminergic neurons in the ventral tegmental area (VTA) is under the influence of GABAergic neurotransmission, and the aim of this study was to characterize the involvement of nAChRs in mediating acute ethanol effects on GABAergic activity in subregions of the brain reward system. Multi-electrode in vivo recordings were made in the VTA and nAc of awake and behaving C57BL6/J mice receiving intraperitoneal injections of saline or ethanol (2.0 g/kg), combined with, or without, pre-injection of the non-competitive nAChR antagonist mecamylamine (1.0 mg/kg). Ethanol significantly decreased the activity of quinpirole-insensitive slow-spiking and fast-spiking units in both the VTA and the nAc as compared to saline injection. Pre-treatment with mecamylamine inhibited the rate-inhibiting properties of ethanol in the VTA, but not in the nAc. The data presented here show that ethanol depresses the activity of quinpirole-insensitive, putative GABAergic neurons, in the mesolimbic dopamine system of mice, and that nAChRs contribute to this modulation. This finding, taken together with previous microdialysis studies, supports an involvement of GABAergic neurons and nAChRs in ethanol's interaction with the mesolimbic dopamine system.     

裙带菜多糖对大鼠黑质内多巴胺能神经元6-羟基多巴胺致损伤的保护作用

目的探讨裙带菜多糖(UPPS)对6-羟基多巴胺(6-OHDA)制备的大鼠帕金森(PD)模型动物的多巴胺(DA)能神经元的保护作用。方法将36只Wistar大鼠随机分为对照组、UPPS+6-OHDA组、6-OHDA组。对照组连续生理盐水灌胃30d;UPPS+6-OHDA组,裙带菜多糖(400mg/kg)灌胃3d后,内侧前脑束(MFB)注射6-OHDA(3.6mg/ml),随后连续裙带菜多糖(400mg/kg)灌胃26d;6-OHDA组,以生理盐水代替裙带菜多糖,其他处理同UPPS+6-OHDA组。检测各组大鼠纹状体(Str)内多巴胺、3,4-二羟苯乙酸(DOPAC)、高香草酸(HVA)的含量,6-OHDA损毁侧大鼠黑质(SN)酪氨酸氢化酶(TH)阳性细胞数和铁染色阳性细胞数,大鼠黑质SOD/β-actin灰度值。结果 UPPS+6-OHDA组DA、DOPAC的含量低于对照组(P相似文献   

Effect of ethanol on ascorbate release in the nucleus accumbens and striatum of freely moving rats.

An in vivo voltammetry technique was used to monitor the extracellular ascorbate (AA) concentration in the nucleus accumbens and striatum of unanesthetized, freely moving rats. A single injection of ethanol, 1.0 g/kg intraperitoneally (IP), induced a significant increase in extracellular AA concentration in both the nucleus accumbens and striatum. This effect was dose dependent within a dose range from 0.5-2.0 g/kg. 4-Methylpyrazole (50 mg/kg, IP), which inhibits alcoholdehydrogenase, could not prevent the increase in AA concentration, evoked by ethanol. Furthermore, systemic administration of acetaldehyde (20 mg/kg, IP), the main metabolite of ethanol, did not have any effect on the level of AA in the nucleus accumbens or striatum. These results show that ethanol can alter the brain extracellular AA levels and that this effect seems to be attributed to ethanol itself and not to acetaldehyde. Consequently, these results indicate that a role for AA in the action of ethanol in the brain should be considered.     

Anatomical "circuitry" in the brain mediating alcohol drinking revealed by THP-reactive sites in the limbic system

The involvement of aldehyde adducts in the etiology of alcoholism continues to be supported by a number of experimental findings. These metabolites are synthesized endogenously from a condensation reaction of a biogenic aldehyde with a catechol- or indole-amine and act in the brain to augment or suppress the drinking of ethyl alcohol. When given by the intracerebroventricular route in an animal which does not prefer alcohol, certain tetrahydro-isoquinolines and beta-carbolines can augment significantly the voluntary intake of alcohol even in aversive concentrations. This paper describes the historical background and current status of the "Multiple Metabolite" theory of alcoholism. The recent identification of anatomical structures in the limbic-midbrain, limbic-forebrain of the Sprague-Dawley rat, which mediate changes in the intake of alcohol induced by tetrahydropapaveroline (THP) is also described. When injected in a low dose of 25 ng in a specific site, over a 3-day period, THP induces persistent increases in the intake of alcohol even in aversive concentrations. These THP-reactive sites comprise the substantia nigra, reticular formation, medial lemniscus, zona incerta, medial forebrain bundle, nucleus accumbens, olfactory tubercle, lateral septal nucleus, preoptic area, stria terminalis, and rostral hippocampus. A higher dose of 250 ng THP microinjected at homologous loci tends to inhibit the rat's self-selection of alcohol or exert no effect on drinking. Morphological mapping of histologically identified sites sensitive to THP revealed a distinct "circuitry" of neuronal structures overlapping both dopaminergic and enkephalinergic pathways. This "circuit" extends from the tegmental-nigral area of the midbrain rostrally to structures within the limbic-forebrain. When a THP-reactive structure, the N. accumbens, was lesioned by either of two neurotoxins, 6-hydroxydopamine or 5.7-dihydroxytryptamine, the rats' preference for alcohol increased sharply. This suggests that impairment of transmitter release, denervation supersensitivity or other perturbation of receptor function within this and other structures play a part in the aberrant drinking of alcohol. It is envisaged that a dopamine-enkephalin link underlies the mechanism for the onset, maintenance and permanency of alcohol preference generated by an aldehyde adduct. Finally, the "Two-Channel, Brain Metabolite" theory of alcoholism proposes that the transitory presence of an endogenously formed aldehyde adduct within cells of the brain causes a permanent perturbation of normal receptor processes and transmitter activity within synapses of specific structures of the limbic system. This theory thus explains the nature of the rewarding properties of alcohol as well as its complex addictive liability which is physiologically irreversible.     

Ethanol intake and ethanol-conditioned place preference are reduced in mice treated with the bioflavonoid agent naringin

Recently, PPAR-γ activation has emerged as a potential treatment for alcoholism. However, the adverse effects of synthetic PPAR-γ activators, despite being effective drugs, prompted the need for novel PPAR-γ agonists that retain efficacy and potency with a lower potential of side effects. Hence, naringin, a bioflavonoid isolated from citrus fruits and recently identified as a natural ligand of PPAR-γ, has begun to be evaluated for treatment of alcoholism. It is well known to possess several therapeutic benefits in addition to its anti-anxiety and antidepressant properties. In the present study, we assessed whether naringin treatment possesses anti-ethanol reward properties in C57BL/6 mice. We used the two-bottle choice drinking paradigm and ethanol-induced conditioned place preference (CPP) to examine the effect of naringin treatment on ethanol drinking. Results have shown that, compared with vehicle, naringin (10–100 mg/kg) significantly and dose-dependently decreased voluntary ethanol intake and preference in a two-bottle choice drinking paradigm [3–15% (v/v) escalating over 2 weeks], with no significant effect observed on saccharin [0.02–0.08% (w/v)] or on quinine [15–60 μM (w/v)] intake. In addition, there was no significant difference in blood ethanol concentration (BEC) between groups following naringin administration of 3 g of ethanol/kg body weight. Interestingly, when mice were treated with vehicle or naringin (30 mg/kg) before injection of ethanol (1.5 g/kg) during conditioning days, naringin inhibited the acquisition of ethanol-CPP. More importantly, these effects were significantly attenuated when mice were pre-injected with the peroxisome proliferator-activated receptor-γ (PPAR-γ) antagonist, GW9662. Taken together, the present findings are the first to implicate naringin and PPAR-γ receptors in the behavioral and reward-related effects of ethanol and raise the question of whether specific drugs that target PPAR-γ receptors could potentially reduce excessive ethanol consumption and preference.     

Buspirone alters alcohol drinking induced in rats by tetrahydropapaveroline injected into brain monoaminergic pathways

The effect of the novel anxiolytic, buspirone, administered systemically was determined in Sprague-Dawley rats induced to drink ethyl alcohol chronically by repeated microinjections of 25 ng/microliter tetrahydropapaveroline HBr (THP) into brain-stem monoaminergic pathways. Self-selection of alcohol in concentrations from 3% to 30% was determined for each rat in a free-choice drinking situation with water available as the alternative fluid. After stereotaxic implantation of guide tubes, THP was microinjected repeatedly into striatal lemniscal and preoptic sites which were found to mediate significant increases in alcohol preference. After the baseline level of intake of a single, maximally preferred alcohol concentration was established, each rat was treated with either saline vehicle or buspirone given intramuscularly b.i.d. in doses of 5.0 mg/kg or 20 mg/kg. Overall, the repeated administration of either dose of buspirone produced a significant decrease in the voluntary alcohol intake of the rats as measured by the proportion measure and absolute g of alcohol ingested. However, the alteration in drinking varied from animal to animal with respect to both magnitude and duration of the anxiolytic's effect. The response to buspirone seemed to be dependent in part on the individual site in each animal at which THP had been infused to evoke alcohol intake. Post-mortem histological analysis revealed that buspirone-treated rats reduced alcohol consumption by 83% if THP had been microinjected into substantia nigra; by 60% if given in the nucleus accumbens-preoptic area; and by 34% when injected into the medial lemniscus-zona incerta. These results suggest, therefore, that buspirone can exert a specific effect in attenuating the consumption of alcohol of the rat in a free-choice situation. In relation to the differential actions of the anxiolytic, it is envisaged that on an anatomical basis the antagonism of alcohol drinking may be mediated by a pharmacological alteration of circumscribed pathways associated with dopaminergic or serotonergic neurons.