Effect of septal damage on alcoholic motivation in the rat

Interconnection between predisposition to depressive-like state (DS) in stress situation and heightened addiction to ethanol was studied in nonlinear male white rats. Electrolytic lesion of the septum in animals inclined to the development of DS, led to a change from the passive type of reaction in stress conditions to the active type. The rate of formation of alcoholic motivation in these animals decreased in comparison with sham-operated rats. The effect of the septal lesion in rats with formed addiction to alcohol was less expressed. Conclusion is made that the passive type of reaction in extreme conditions is significant for the formation of alcohol motivation in rats, especially at early stages.     

Effects of passive administration of serotonin and catecholamine antibodies on alcohol consumption of C57Bl/6 mice in experimental alcoholism]

The experiments on C57Bl/6 mice with natural ethanol motivation have shown that passive injection of anti-noradrenaline and particularly antiserotonin antibodies reduces several times alcohol consumption by animals. On the contrary the injection of anti-dopamine antibodies to affect voluntary alcohol consumption by mice have been found.     

The effect of valproic acid on sleep structure and ethanol consumption in rats with various types of individual reactivity before and after stress exposure

The influence of valproic acid on sleep structure and alcohol motivation was studied in pretyped rats prior and following REM sleep deprivation. During EEG recording of wake-sleep cycle valporoic acid was shown to produce hypnotic action dependent on drug dosage in high active and low active animals. There was also shown that following REM sleep deprivation low active animals significantly reduced ethanol consumption under valproic acid influence. It seems likely from the results obtained that valproic acid could be used as hypnotic and antialcoholic drug.     

Coenzyme binding in alcohol dehydrogenase

Crystallographic investigations of horse liver alcohol dehydrogenase have demonstrated that NAD is not a passive participant in the redox reactions catalysed by the enzyme. On the molecular level NAD acts as an activator which induces an active form of the enzyme. This is mediated by a large conformational change, making the active site dehydrated and by providing one part of the substrate-binding cleft. The catalytic events, substrate binding, inhibitor binding and the role of the catalytic zinc ion are discussed in relation to the role of NAD. Human alcohol dehydrogenase isoenzymes which have very different substrate specificities are discussed in relation to sequence differences.     

Lack of lead effects on fetal development and offspring learning when combined with alcohol in the Long-Evans rat

Two studies were conducted to evaluate the interactive effects of alcohol and lead during pregnancy in rats. Our purpose was to see if lead, as lead acetate, would influence the alcohol effect already known to exist. In the first study, pregnant Long-Evans rats received lead (as lead acetate), alcohol (20% w/v), or lead plus alcohol once a day on gestation days (GD) 10-20. On GD 20, when animals were sacrificed, mean blood alcohol levels were consistently higher for the lead-plus-alcohol-dosed groups compared to alcohol alone, but these two groups did not differ in maternal weight gain, percent resorptions, litter size, or fetal weight. Mean blood lead levels were not consistently higher in the lead-plus-alcohol groups compared to lead only, but the lead-plus-alcohol groups differed significantly from the lead-only groups at higher doses in the previously mentioned parameters. The lead-only groups did not differ from vehicle controls in any parameter in spite of blood lead levels as high as 300 micrograms/dl. In the second experiment, animals given a combination of alcohol and lead did not differ in activity, passive avoidance, or active avoidance learning compared to animals given alcohol or lead alone. Animals given lead only or the combination of lead plus alcohol had longer first trial latencies in the passive avoidance test. The data indicate that neither lead nor alcohol attenuates or potentiates each other's effects on reproduction or learning behavior in the Long-Evans rat even at high blood lead levels.     

Effect of psychotropic substances on the development of alcoholic motivation in noninbred white rats

The experiments have shown the capacity of antidepressant amitriptylin (0.5 mg/kg, i. p.) and tranquilizer phenazepam (0.1 mg/kg i. p.) to normalize the adaptive behaviour and almost completely prevent the development of alcohol motivation in animals with insufficient adaptive behaviour. It was established that in animals initially rejecting alcohol, chronic treatment with these drugs as well as d-amphetamine promoted alcohol motivation. The results obtained have proved our earlier hypothesis that preclinical search for drugs for the prevention and treatment of early stages of alcoholism is possible only in animals pre-selected according to their inclination to experimental alcoholism.     

Effect of the destruction of the brain serotoninergic system on the alcohol consumption by rats in the early periods of experimental alcoholism

Albino noninbred rats were divided into groups, according to the duration of alcoholic anesthesia (4.5 g/kg i.p.), of predisposed (195.6 min) and non-predisposed (69.1 min) to voluntary intake of alcohol. Another group included animals screened for 21 days according to the level of intake of 15% ethanol under the conditions of free choice between alcohol and water (6.15 and 2.62 g/kg pure ethanol per day, respectively). The animals were subjected to electro-coagulation of the dorsal or magnus raphe nucleus or were injected with 5,6-dihydroxytryptamine--DNT (75 micrograms/microliters) into the ventricles of the brain. It was established that in rats non-predisposed to alcohol intake, the destruction of the raphe nuclei, of the dorsal in particular, or injection of DOT to animals with a weak alcoholic motivation produces a dramatic increase in alcohol intake. In alcohol intake predisposed rats and in animals with a high level of alcohol use, analogous exposures do not bring about any significant differences in alcohol intake. The data obtained indicate that the reduced serotonin content in the brain is associated with an increase in the level of alcoholic motivation.     

The effect of ethanol on zinc and copper metabolism in rats

Studies performed on adult female rats over a period of 10 weeks indicated that the consumption of alcohol (20% v/v) did not appear to disturb the zinc or copper balance, nor did it adversely affect tissue zinc or copper levels, even in zinc-restricted animals. On the contrary, higher plasma zinc levels were consistently observed in animals receiving alcohol together with the experimental diets.     

Effect of ethanol on the concentration of serotonin in the brain of the rat and the excretion of 5-hydroxyindoleacetic acid

It is found that serotonin content in the brain areas and heart of rats with low alcohol motivation decreases after 5 months of chronic consumption of 48% ethanol solution in a dose of 4 g/kg; in animals with high alcohol motivation serotonin content decreases only in the hypothalamus. Under chronic alcoholization for 1 and 12 months no considerable changes were found in serotonin level of the studied tissues. 60 min after intraperitoneal administration of 20% ethanol solution in a dose of 3 g/kg in intact animals there occurs an increase of serotonin content in the brain hemispheres and heart and its decrease in the hypothalamus; in rat with low alcohol motivation after taking ethanol for 5 months this administration evokes a decrease of serotonin content in the hypothalamus and truncus cerebri; in rats with high alcohol motivation--its decrease in the hypothalamus. Excretion of 5-oxyindoleacetic acid with urine decreases 10 months after alcohol intoxication. When rats were not given ethanol after its chronic taking for 3 months serotonin oxidation was intensified for the first day, which was not observed after 7-month alcoholization of animals.     

Behavioral models of the relation of animals to alcohol

Worked out on the outbred white male rats the behavioural models allow to pick out animals with initial alcohol motivation from the intact rats' group and to pick out animals with alcohol dependency and withdrawal syndrome from chronically alcoholized rats' group.     

A comparative evaluation of alcoholic motivation in animals by 2 behavioral methods]

The attitude of outbred white male rats to alcohol was determined by means of two behavioural methods--the method of free choice of alcohol or water and of reaction of preference of place (PP) connected with taking of alcohol. Conclusion is made about greater adequacy of PP application for determination of alcohol motivation in animals.     

Influence of emotional stimuli on behavior and respiration of rabbits with various locomotor activity in the "open field"

Different behavioral reactivity of rabbit groups differentiated by locomotor activity in the "open field" was revealed during exposure to emotional stimuli (rustle, loud sound, pressuring on back of the neck, vibroacoustic tactile stimulation of an ear). In passive rabbits, the active locomotor reactions were induced harder and freezing was obtained easier than in active animals. During exposure to sound stimuli, passive rabbits increased their locomotion more rarely than active animals, pressing on back of the neck produced longer freezing, a threshold of defensive ear shaking in response to a vibroacoustic stimulus in passive animals was highest. Training to mild immobilization increased the threshold of defensive responses in active rabbits and animals of the intermediate type. Changes in respiratory parameters were correlated with behavioral reactions to emotional stimuli. The duration of exhalation and respiratory cycle increased during freezing and increased during enhanced locomotion. The duration of inhalation decreased in response to emotional stimuli irrespective of a behavioral reaction. The respiratory reactions to emotional stimuli differed in rabbits of different groups. The respiratory rate more frequently changed in passive rabbits than in animals of other groups. Passive animals reacted mainly by exhalation, active rabbits and animals from the intermediate group predominantly responded by inhalation.     

Correlation between the rate of ethanol elimination and the psychophysiological characteristics of rats

Ethanol elimination from the blood of rats with different psychophysiological features was studied using gas chromatographic head-space analysis in the general complex of tests aimed at determination of ethanol consumption. The selection of animals with different levels of the initial alcohol motivation was performed according to modified Porsolt's method. It was shown that the initial level of predisposition to depression-like states is in a dose-dependent correlation with the high rate of ethanol elimination. This is suggested to be one of the genetic indications which promotes the formation of the initial alcohol motivation and the development of experimental alcoholism.     

Interaction of amygdalar neurons of active and passive rabbits in negative emotional situations

Interaction of basal and central nuclear neurons of amygdala was studied by plotting histograms of crosscorrelation in passive and active rabbits exposed to emotionally significant stimuli. The behavior of animals was studied in the open field, light-dark test and during presentation of emotionally significant stimuli. Rabbits of different typological groups applied a certain behavioral strategy in a variety of behavioral tests. Intergroup differences were revealed in the interaction of neighboring cells of amygdala. Passive rabbits (as comparied to active rabbits) demonstrated more excitatory connections and less inhibitory connections with the latency from 50 to 150 ms. Interactions with the delta1-range and theta2-range frequencies in passive animals were more rarely observed. The asymmetry of the interhemispheric neuronal interaction in amygdala with the right dominance was revealed in passive but not active animals. The results testify that amygdala is involved in the choice of behavioral strategy, and the level of its activation is higher in passive animals.     

Sorbitol dehydrogenase is a zinc enzyme.

Evidence is given that tetrameric sorbitol dehydrogenase from sheep liver contains one zinc atom per subunit, most probably located at the active site, and no other specifically bound zinc or iron atom. In alcohol dehydrogenases that are structurally related to sorbitol dehydrogenase, more than one zinc atom per subunit can complicate investigations of zinc atom function. Therefore, sorbitol dehydrogenase will be particularly valuable for defining the precise roles of zinc in alcohol and polyol dehydrogenases, and for establishing correlations of structure and function with other important zinc-containing proteins.     

Accumulation of zinc and cadmium by Cytophaga johnsonae

Passive and active accumulation of zinc and cadmium by a common soil and freshwater bacterium, Cytophaga johnsonae, was studied using a radio-tracer batch distribution technique. The effects of variation of pH (3–10), as well as of ionic strength (0.007 and 0.07 m) on passive accumulation of the metals were examined. For both zinc and cadmium, accumulation was mainly due to passive processes, such as surface adsorption and/or diffusion into the periplasm. However, at low zinc concentrations, accumulation increased when glucose was added, suggesting an active uptake; at higher zinc concentrations such uptake was not detected, probably because it was masked by the stronger sorption properties of the cell wall. Adsorption of the metals was pH dependent: at higher ionic strength, accumulation was enhanced at pH values above 7; at lower ionic strength, adsorption differed and was markedly higher, with increased accumulation between pH 5 and 8.     

Human glutathione-dependent formaldehyde dehydrogenase. Structures of apo,binary, and inhibitory ternary complexes

The human glutathione-dependent formaldehyde dehydrogenase is unique among the structurally studied members of the alcohol dehydrogenase family in that it follows a random bi bi kinetic mechanism. The structures of an apo form of the enzyme, a binary complex with substrate 12-hydroxydodecanoic acid, and a ternary complex with NAD+ and the inhibitor dodecanoic acid were determined at 2.0, 2.3, and 2.3 A resolution by X-ray crystallography using the anomalous diffraction signal of zinc. The structures of the enzyme and its binary complex with the primary alcohol substrate, 12-hydroxydodecanoic acid, and the previously reported binary complex with the coenzyme show that the binding of the first substrate (alcohol or coenzyme) causes only minor changes to the overall structure of the enzyme. This is consistent with the random mechanism of the enzyme where either of the substrates binds to the free enzyme. The catalytic-domain position in these structures is intermediate to the "closed" and "open" conformations observed in class I alcohol dehydrogenases. More importantly, two different tetrahedral coordination environments of the active site zinc are observed in these structures. In the apoenzyme, the active site zinc is coordinated to Cys44, His66 and Cys173, and a water molecule. In the inhibitor complex, the coordination environment involves Glu67 instead of the solvent water molecule. The coordination environment involving Glu67 as the fourth ligand likely represents an intermediate step during ligand exchange at the active site zinc. These observations provide new insight into metal-assisted catalysis and substrate binding in glutathione-dependent formaldehyde dehydrogenase.     

Peculiarities of Development of the Depression State in Rats Characterized by Different Individual/Typological Behavioral Statuses

We examined the development of a depression state induced by long-lasting stressing of rats preliminarily selected according to their active or passive behavior pattern. Stress was induced using a technique of sensory contact with aggressive individuals. The pathologically modified state in animals of the above groups developed according to dissimilar scenarios. As a result, a depressive state similar to anxious depression developed in active animals, while psychoemotional disorders similar to a melancholy-type depression were formed in passive rats. Therefore, the symptomatic heterogeneity of depression is, to a considerable extent, determined by individual/typological peculiarities of behavior (passive vs active) and can be rather clearly identified using the sensory contact technique.     

Changes in autoregulation of coronary blood flow in rats with various sensitivity to stress

The purpose of work: study of a role of endothelial nitric oxide in development of stress-induced changes in autoregulation of coronary blood flow in rats with various types of behaviour. The experiments were performed on isolated hearts of female-rats, in the "open field" test, depending on the type of impelling and searching activity of animals subdivided into two groups: "active" and "passive". After a 6-hour immobilization stress only in "passive" rats an increase of volumetric velocity of coronary flow; a decrease of an autoregulation index, coronary reserve against the background of intravascular pressure reduction, were found out. The blockade of nitric oxide synthesis in this group completely eliminated the stress-induced decrease of coronary vascular tone and essentially limited the caused by stress dissociation of coronary flow and the contractility function of the myocardium. In blood plasma of "passive" animals the nitrite/nitrate contents was by 55% more than of the "active" rats. After the transferred stress, in "passive" animals the nitrite/nitrate concentration in blood plasma increased by 29% and in "active" rats--by 136%; the absolute values, however, did not differ between the groups. Thus the autoregulation of coronary flow seems to be subject to action of stress in the rats showing a "passive" type of behaviour in the test "open field", and practically does not change in "active" animals; secondly, in spite of the fact that the stress-induced amplification of NO-producing function andothelium of coronary blood vessels is stereotyped in different rats, in "passive" rats, apparently, sensitivity of coronary vessels to nitric oxide is higher than at "active" those.