Aapo Honkanen

Alcohol drinking is reduced by a μ1- but not by a δ-opioid receptor antagonist in alcohol-preferring rats

To assess the roles of opioid receptor subtypes in voluntary alcohol drinking, alcohol-preferring AA (Alko, Alcohol) rats, non-deprived of food or water, were used in a paradigm where access to 10% alcohol solution was limited to 1–4-h sessions on every 2nd working day. The δ-opioid receptor antagonist naltrindole (1–5 mg/kg i.p. 15 min before the session) had no effect on alcohol drinking, while it attenuated the δ-opioid receptor agonist [d-Pen2,d-Pen5]enkephalin-induced locomotor stimulation. The μ1-opioid receptor antagonist naloxonazine (1–15 mg/kg i.p. 20 h before the session), at the largest dose, decreased alcohol drinking. It also decreased food intake. When naltrindole (1 mg/kg) and naloxonazine (15 mg/kg) were given prior to 3 consecutive sessions, the former had no effects at any session. Naloxonazine decreased alcohol consumption only in the 1st session, although the reduction of daily water intake became stronger during repeated administration. 4 days after the last drug administration, naloxonazine-treated animals consumed alcohol nearly twice as much as in the control session before any drug treatment. These data suggest that δ-opioid receptors are not involved in the regulation of alcohol drinking in AA rats. μ1-Opioid receptors may be involved in alcohol drinking, although the data suggest that even their prolonged blockade alone is insufficient to induce a sustained decrease in alcohol drinking.

Benzodiazepine receptor ligands modulate ethanol drinking in alcohol-preferring rats

The effects of benzodiazepine receptor ligands with different intrinsic activity profiles were studied on voluntary ethanol consumption in the selectively bred alcohol-preferring AA (Alko, Alcohol) rat line, and compared to those of an opiate antagonist, naloxone, and a serotonin uptake inhibitor, citalopram. The rats were first allowed to develop a strong preference for 10% (v/v) ethanol solution in tap water over plain water until their ethanol consumption stabilized. Thereafter, the period when ethanol solution was available for the rats was gradually reduced to 4 h, 3 times a week, every second working day. The acute effects of positive allosteric modulators (agonists) of the gamma-aminobutyric acid type A (GABAA)/benzodiazepine receptor [midazolam, abecarnil, ethyl 5-benzyloxy-4-methoxymethyl-beta-carboline-3-carboxylate (ZK 91296), bretazenil, and 2,5-dihydro-2-(4-methylphenyl)-3H-pyrazolo[4,3-C]quinolin-3(5H)-on e (CGS 9895)] and of negative allosteric modulators [inverse agonists, ethyl 8-azido-5,6-dihydro-5-methyl-6-oxo-4H-imidazo[1,5- a][1,4]benzodiazepine-3-carboxylate (Ro 15-4513) and t-butyl 5,6-dihydro-5-methyl-6-oxo-4H-imidazo[1,5-a]thieno[2,3- f][1,4]diazepine-3-carboxylate (Ro 19-4603)] were tested after i.p. injections of three different drug doses using saline injections as a control treatment. The benzodiazepine agonists had rather modest effects on ethanol intake, measured 1 and 4 h after the injections, whereas the inverse agonists and naloxone strongly decreased ethanol consumption. Acute citalopram had no clear effect on ethanol drinking, but it slightly decreased the consumption of novel food during the 4-h session, as did all other benzodiazepine agonists except bretazenil. Neither the inverse agonists nor naloxone had any significant effect on food intake.(ABSTRACT TRUNCATED AT 250 WORDS)

[3H]Ethylketocyclazocine Binding to Brain Opioid Receptor Subtypes in Alcohol-Preferring AA and Alcohol-Avoiding ANA Rats

We measured brain regional patterns of [3H]ethylketocyclazocine binding to brain opioid receptors in ethanol-naive alcohol-preferring Alko, Alcohol (AA) and alcohol-avoiding Alko, Non-Alcohol (ANA) rats, by using quantitative autoradiography. This agonist ligand labels all opioid receptor subtypes. The proportions of mu- and delta-opioid receptor binding were evaluated by displacing the mu- and delta-opioid receptor components by the peptides Tyr-D-Ala-Gly-N(Me)Phe-Gly-ol (DAMGO, 100 nM) and Tyr-D-Pen-Gly-Phe-D-Pen (DPDPE, 100nM), respectively, the K-component being the naltrexone-sensitive binding left after removal of the above two components. The labeling patterns in the brains of the AA and ANA rats were consistent with the well-known distributions of the opioid receptor subtypes in nonselected rat strains and there was no major difference between the lines. The mu-opioid receptor binding was greater in the AA than ANA rats in several brain regions, most interestingly in the substantia nigra pars reticulata and striatal clusters with elevated shell/core ratios in the nucleus accumbens. The delta-opioid receptor binding did not differ between the lines, whereas the AA rats had more K-opioid receptors than the ANA rats in several brain regions, including limbic areas and basal ganglia. The observed results might indicate altered action of the opioidergic system on dopaminergic pathways in rats with differential alcohol preference.

Brain Opioid Receptor Binding of [3H]CTOP and [3H]Naltrindole in Alcohol-Preferring AA and Alcohol-Avoiding ANA Rats

We compared mu- and delta-opioid receptor distributions between the brains of alcohol-preferring Alko, Alcohol (AA) and alcohol-avoiding Alko, Non-Alcohol (ANA) rat lines, using autoradiography on brain sections with mu- and delta-opioid receptor antagonist ligands [3H]CTOP and [3H]naltrindole, respectively. The labeling patterns of the ligands were consistent with the known opioid receptor distributions in both rat lines and no major genetic differences were found between the lines. However, the binding density of mu- and delta-opioid receptors differed slightly in several brain areas: in the AA brain sections, limbic areas, such as hippocampus and amygdala, showed decreased mu- and delta-opioid receptor binding, whereas the striatal patches were larger and the substantia nigra showed higher binding density of the mu-receptors compared to the ANA sections. The small differences observed between the rat lines could be due to adaptations to altered endogenous opioid peptide levels or neural circuits, and associated with the differences in alcohol drinking or other behaviors.

Involvement of Opioid μ1-Receptors in Opioid-Induced Acceleration of Striatal and Limbic Dopaminergic Transmission

The role of mu1-opioid receptors in the acceleration of cerebral dopaminergic transmission induced by morphine and the putative mu1-opioid agonist, etonitazene, was studied in rats by measuring the tissue levels of dopamine (DA) and its metabolites 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) in the dorsal striatum and nucleus accumbens. The striatal extracellular concentrations of DA and its metabolites in freely moving rats were estimated as well. Morphine (3 mg/kg) and etonitazene (2.5 microg/kg) increased the striatal and accumbal dopamine metabolism as measured by the tissue ratios of DOPAC/DA and HVA/DA. The mu1-opioid receptor antagonist, naloxonazine (15 mg/kg), significantly antagonized these elevations except the morphine-induced elevation of striatal HVA/DA ratio. Both morphine (3 mg/kg) and etonitazene (1, 2.5, and 5 microg/kg) elevated the striatal extracellular DA, DOPAC, and HVA. Naloxonazine antagonized the effects of morphine and etonitazene on striatal extracellular DA concentration as well as etonitazenes effects on DOPAC and HVA, but not morphines effects on DOPAC and HVA. As we previously showed concerning morphine, the conditioned place preference induced by etonitazene was inhibited by naloxonazine. These findings emphasize the role of mu1-opioid receptors in opioid reward, in which the mesolimbic dopaminergic system is considered to be importantly involved. Our results clearly show that in addition to the mesolimbic dopaminergic system the mu1-opioid receptors are also involved in the control of nigrostriatal DA release and metabolism. However, the effects of etonitazene on the striatal DA differ from those of morphine, suggesting that the opioid mechanisms regulating these two DA systems differ.

Morphine-stimulated metabolism of striatal and limbic dopamine is dissimilarly sensitized in rats upon withdrawal from chronic morphine treatment

The effects of acute morphine on the release of dopamine (DA) in the striatum and limbic forebrain of rats upon 48 h withdrawal from 20-day morphine treatment were studied using 3-methoxytyramine (3-MT) in tissue as an index of DA release. Homovanillic acid (HVA) and 3,4-dihydroxyphenylacetic acid (DOPAC) were also measured. The chronic morphine treatment did not alter the concentrations of DA metabolites. Acute morphine (10 mg/kg) elevated all three DA metabolites in both brain areas. Morphine withdrawal potentiated the elevation of striatal and limbic 3-MT as well as that of striatal but not limbic HVA. These findings show that both striatal and limbic DA mechanisms are sensitized to morphine upon withdrawal but that sensitization of DA metabolism in these two brain areas occurs differently.

Effects of Morphine on Metabolism of Dopamine and Serotonin in Brains of Alcohol-Preferring AA and Alcohol-Avoiding ANA Rats

Morphine induces a larger locomotor stimulation in the alcohol-preferring AA rats than in the alcohol-avoiding ANA rats. We have now studied the acute effects of morphine (1 and 3 mg/kg) on metabolism of dopamine and serotonin (5-HT) in the dorsal and ventral striatum of the AA and ANA rats. The basal level of dopamine release, as reflected by the concentration of dopamine metabolite 3-methoxytyramine (3-MT), was lower in the caudate-putamen and nucleus accumbens of the AA rats than in the ANA rats. In the caudate-putamen, morphine increased dopamine metabolism and release more in the AA than in the ANA rats. In the nucleus accumbens and olfactory tubercle, the effects of morphine on dopamine metabolism and release did not differ between the rat lines. Morphine elevated the metabolism of 5-HT in the caudate-putamen and nucleus accumbens of the AA but not in those of the ANA rats. The results suggest that the larger morphine-induced psychomotor stimulation of the AA rats in comparison with the ANA rats is associated with the larger effect of morphine on dopamine metabolism in the caudate-putamen and 5-HT metabolism in the caudate-putamen and nucleus accumbens. Furthermore, low basal dopamine release may play a role in the high alcohol-preference of AA rats.

Conditioned place preference induced by a combination of L-dopa and a COMT inhibitor, entacapone, in rats

The interaction of dopamine (DA) precursor L-dopa and catechol-O-methyltransferase (COMT) inhibitor, entacapone, was examined in rats using conditioned place preference (CPP) paradigm to assess reinforcement, and by measuring DA metabolism in the striatum and the limbic forebrain. Neither L-dopa (100 mg/kg i.p.) nor entacapone (30 mg/kg i.p.) alone induced CPP, but in combination they induced significant CPP. Entacapone alone had no effect on limbic or striatal DA concentrations, while it reduced the concentrations of the COMT products 3-methoxytyramine (3-MT), a metabolite reflecting DA release, and homovanillic acid (HVA) in both brain areas. L-dopa elevated limbic but not striatal 3-MT. L-dopa also slightly elevated limbic DA but had no effect on striatal DA concentration. L-Dopa-induced increase of 3-MT was attenuated by entacapone. Our results show for the first time that L-dopa is able to produce CPP in intact animals. This effect may be related to the findings that L-dopa increases synaptic DA concentrations in the limbic areas, and entacapone may enhance this elevation as it prevents the synaptic metabolism of DA.

Voluntary alcohol drinking selectively accelerates dopamine release in the ventral striatum as reflected by 3-methoxytyramine levels

Alcohol-preferring Alko, Alcohol (AA) rats with free access to food and water were taught to voluntarily consume 10% alcohol solution during 20-min sessions. After the drinking session, rats had elevated dopamine release (as reflected by 3-methoxytyramine concentrations) in the nucleus accumbens, olfactory tubercle, and caudate-putamen. When alcohol solution was replaced by plain water, dopamine release was increased only in the caudate-putamen, indicating selective activation of the mesolimbic dopaminergic pathway by alcohol.

Effects of repeated morphine on cerebral dopamine release and metabolism in AA and ANA rats

Cerebral dopaminergic mechanisms were studied in the nucleus accumbens and caudate-putamen of alcohol-preferring AA (Alko Alcohol) and alcohol-avoiding ANA (Alko Non-Alcohol) rats after 4-day repeated morphine treatment. This treatment has been shown to enhance the locomotor activity stimulating effect of morphine in the AA but not in the ANA rats. Morphine (1 or 3 mg/kg) or saline was administered subcutaneously once daily and the extracellular concentrations of dopamine (DA), 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) were measured, in freely moving rats by in vivo microdialysis on days 1 and 4. Morphine increased accumbal DA, DOPAC and HVA similarly in rats of both lines, and no sensitization of DA release or metabolism was seen in rats of either line given morphine repeatedly. In the caudate-putamen, morphine increased DA, DOPAC and HVA significantly only in the AA rats. During repeated treatment, the morphine-induced elevation of DA metabolites, but not that of DA, was enhanced similarly in rats of both lines. These results suggest that the effects of acute morphine administration on nigrostriatal dopaminergic mechanisms are stronger in the AA than in the ANA rats, whereas the effects of morphine on mesolimbic dopaminergic systems do not differ. Furthermore, in rats of both lines, repeated morphine treatment enhanced the responses of the nigrostriatal dopaminergic systems similarly, but no enhancement occurred in the mesolimbic systems of rats of either line. These findings do not support the critical role of accumbal dopaminergic systems in morphine-induced behavioural sensitization.