• Energy Research

Our previous findings have shown that l-cysteine, a non essential amino acid, prevented ethanol (EtOH) induced conditioned place preference. The aim of the present study was to examine the effect of l-cysteine on the acquisition and maintenance of oral EtOH self-administration and on the reinstatement of EtOH-drinking behavior in Wistar rats. Rats were pretreated intraperitoneally with saline or l-cysteine (20 and 40 mg/kg) 30 min before each acquisition trial, in an operant nose-poking paradigm where they were given the opportunity to orally self-administer tap water or EtOH (5-10% v/v). Further, to evaluate if l-cysteine reduces the acquired oral EtOH self-administration, we carried out an independent experiment in which rats were trained to self-administer EtOH (10%); after all groups of rats developed similarly stable oral EtOH self-administration, the effect of l-cysteine (0, 40, 60, 80 and 100mg/kg) was tested. An additional group of rats was pretreated with saline or l-cysteine (80 mg/kg) and tested on reinstatement after EtOH extinction and, at the end of last reinstatement session, were utilized to measure blood and brain EtOH levels. The animals that had access to EtOH solution discriminated between the active and inactive nose-pokes and showed rates of active nose-pokes significantly higher than the tap water group. Furthermore, rats self-administering EtOH (10%) also demonstrated extinction behavior and gradually reinstated active nose-poke responding when EtOH was reintroduced. l-cysteine reduced both the acquisition and maintenance of oral EtOH self-administration. The reduced reinstatement of EtOH-drinking behavior was paralleled by a significant reduction of EtOH intake and correlated with blood and brain EtOH levels. The efficacy of l-cysteine on the various phases of alcohol drinking in rats, could represent an interesting pharmacological approach and could open a new line of research for the development of therapies to reduce EtOH intake in alcoholic patients.

Rats maintained for 10 days on a 10% ethanol solution as the sole source of fluid developed marked tolerance to ethanol-induced loss of righting reflex, but no tolerance to the stimulatory effects on mesolimbic dopaminergic system. An ethanol challenge stimulated both the electrical activity of A10 dopaminergic cells and dopamine output in the ventral striatum of behaviourally tolerant animals and of controls to the same extent. These results are compatible with the hypothesis that an increased dopamine neurotransmission in the limbic system participates in the reinforcing effect of ethanol.

AbstractEthanol (EtOH), the main psychoactive ingredient of alcoholic drinks, is widely considered to be responsible for alcohol abuse and alcoholism through its positive motivational properties, which depend, at least partially, on the activation of the mesolimbic dopaminergic system. However, acetaldehyde (ACD), the first metabolite of EtOH, has been classically considered to be aversive and useful in the pharmacological therapy of alcoholics. Here we show that EtOH‐derived ACD is necessary for EtOH‐induced place preference, a pre‐clinical test with high predictive validity for reward liability. We also found that ACD is essential for EtOH‐increased microdialysate dopamine (DA) levels in the rat nucleus accumbens and that this effect is mimicked by intra‐ventral tegmental area (VTA) ACD administration. Furthermore, in vitro, ACD enhances VTA DA neuronal firing through action on two ionic currents: reduction of the A‐type K+ current and activation of the hyperpolarization‐activated inward current. EtOH‐stimulating properties on DA neurons are prevented by pharmacological blockade of local catalase, the main metabolic step for biotransformation of EtOH into ACD in the central nervous system. These results provide in‐vivo and in‐vitro evidence for a key role of ACD in the motivational properties of EtOH and its activation of the mesolimbic DA system. Additionally, these observations suggest that ACD, by increasing VTA DA neuronal activity, would oppose its well‐known peripherally originating aversive properties. Careful consideration of these findings could help in devising new effective pharmacological therapies aimed at reducing EtOH intake in alcoholics.

Ethyl alcohol (EtOH), the main psychoactive ingredient of alcoholic drinks, is widely considered responsible for alcohol abuse and alcoholism through its positive motivational properties, which depend, at least partially, on the activation of the mesolimbic dopaminergic system. On the other hand, acetaldehyde (ACD), EtOH's first metabolite, has been classically considered aversive and useful in the pharmacologic therapy of alcoholics. Here we show that EtOH‐derived ACD is necessary for EtOH‐induced place preference, a preclinical test with high predictive validity for reward liability. We also found that ACD is essential for EtOH‐increased microdialysate dopamine (DA) levels in the nucleus accumbens (NAcc), and that this effect is mimicked by ACD administration to the intraventral tegmental area (VTA). Furthermore,in vitro, ACD enhances VTA DA neuronal firing. Coherently, EtOH‐stimulating properties on DA neurons are prevented by pharmacologic blockade of local catalase: the main metabolic step for biotransformation of EtOH into ACD in the central nervous system. These results providein vivoandin vitroevidence for a key role of ACD in EtOH motivational properties and its activation of the mesolimbic DA system. Additionally, these observations suggest that ACD, by increasing VTA DA neuronal activity, would oppose its well‐known peripherally originating aversive properties. These findings could help in devising new effective pharmacologic therapies in alcoholism.

The effect of ethanol challenge on the extracellular concentrations of dopamine, 3,4-dihydroxy-phenylacetic acid and homovanillic acid was studied in the ventral striatum of rats repeatedly treated with ethanol. Ethanol-treated animals (1 g/kg i.p. twice a day for 12 days) developed marked tolerance to the behavioral signs of ethanol intoxication when challenged with ethanol (2 g/kg i.p.). However, in ethanol-treated animals the increased output of dopamine and metabolites after ethanol challenge (1 or 2 g/kg i.p.) was not statistically different from that observed in saline-treated rats. These results indicate that tolerance does not develop to the ethanol-induced stimulation of dopamine release and support the hypothesis that activation of the mesolimbic dopamine system contributes to the reinforcing properties of ethanol.

Background:  Acetaldehyde (ACD), the first metabolite of ethanol (EtOH), is produced peripherally by gastric and hepatic alcohol dehydrogenase (ADH) and centrally by brain catalase. In spite of the aversive properties classically ascribed to ACD, it has recently been suggested that ACD might mediate some of the motivational effects of EtOH. Accordingly, the relative role of ACD in the positive motivational properties of EtOH ingested is increasingly becoming the matter of debate. Thus, we studied the ability of intragastrically administered EtOH, ACD and EtOH‐derived ACD to induce conditioned place preference (cpp) in rats.Methods:  Wistar rats were pretreated intraperitoneally with saline, the peripheral competitive inhibitor of ADH, 4‐methylpyrazole (4‐MP, 22.5, 45 or 67.5 mg/kg) or with the selective ACD‐sequestrating agent, d‐penicillamine (DP, 25 or 50 mg/kg), before the intragastric administration of saline, EtOH (0.5, 1 or 2 g/kg) or ACD (10, 20, or 40 mg/kg). The specificity of 4‐MP and DP effects was addressed using morphine‐induced cpp (2.5 mg/kg).Results:  Both, EtOH and ACD dose‐dependently induced cpp; further, while EtOH‐induced cpp was prevented by the administration of 4‐MP and by DP, ACD‐induced cpp was unaltered by 4‐MP administration and prevented by DP. Both pretreatments did not interfere with morphine‐induced cpp indicating that 4‐MP and DP specifically modulate the motivational properties of EtOH and ACD.Conclusion:  The ability of 4‐MP and DP to decrease EtOH‐induced cpp suggests that a reduction of ACD levels is crucial in depriving EtOH from its motivational properties as indexed by the cpp procedure. In addition, this conclusion is supported by the inefficacy of 4‐MP in preventing ACD‐induced cpp, and by its blockade observed after administration of the selective ACD sequestrating agent DP. The present results underscore the role of EtOH‐derived ACD in EtOH‐induced motivational properties as well as its abuse liability.

Rats chronically administered with ethanol every six hours for six consecutive days show, upon suspension of treatment, a marked somatic withdrawal syndrome characterized by classical neurological signs. The emergence of the behavioral syndrome coincides with a profound decline of dopaminergic mesolimbic neuronal activity which corresponds to a reduction of dopamine outflow in the nucleus accumbens [Diana et al. (1993) Proc. natn. Acad. Sci. U.S.A. 90, 7966-7969]. However, while the behavioral manifestation of the ethanol withdrawal syndrome recedes in about 48 h, electrophysiological indices of mesolimbic dopaminergic function are still reduced 72 h after ethanol discontinuation, thus outlasting the physical signs of ethanol withdrawal syndrome. Dopaminergic neuronal activity is reintegrated by anti-craving drugs such as ethanol itself and gamma-hydroxybutyric acid. It is postulated that the reduced spontaneous activity of mesolimbic dopaminergic neurons may form the neural basis of the dysphoric state which accompanies abrupt interruption of chronic ethanol administration. Pharmacological manipulations of dopaminergic activity targeted at restoring "normal" dopaminergic function after ethanol withdrawal may lead to way to the experimental basis of new therapeutic strategies of alcoholism.

Acetaldehyde (ACD), the first metabolite of ethanol, has been implicated in several behavioural actions of alcohol, including its reinforcing effects. Recently, we reported that l-cysteine, a sequestrating agent of ACD, reduced oral ethanol self-administration and that ACD was orally self-administered. This study examined the effects of l-cysteine pre-treatment during the acquisition and maintenance phases of ACD (0.2%) self-administration as well as on the deprivation effect after ACD extinction and on a progressive ratio (PR) schedule of reinforcement. In a separate PR schedule of reinforcement, the effect of l-cysteine was assessed on the break-point produced by ethanol (10%). Furthermore, we tested the effect of l-cysteine on saccharin (0.2%) reinforcement. Wistar rats were trained to self-administer ACD by nose poking on a fixed ratio (FR1) schedule in 30-min daily sessions. Responses on an active nose-poke caused delivery of ACD solution, whereas responses on an inactive nose-poke had no consequences. l-cysteine reduced the acquisition (40 mg/kg), the maintenance and the deprivation effect (100 mg/kg) of ACD self-administration. Furthermore, at the same dose, l-cysteine (120 mg/kg) decreased both ACD and ethanol break point. In addition, l-cysteine was unable to suppress the different responses for saccharin, suggesting that its effect did not relate to an unspecific decrease in a general motivational state. Compared to saline, l-cysteine did not modify responses on inactive nose-pokes, suggesting an absence of a non-specific behavioural activation. Taken together, these results could support the hypotheses that ACD possesses reinforcing properties and l-cysteine reduces motivation to self-administer ACD.