The impact of background γ-radiation on erythrocyte nuclear pathology, the serotonergic system, and cytochrome P-450 in hens (GALLUS GALLUS DOMESTICUS) from Azerbaijan.

High levels of background γ-radiation exist in the suburbs of Baku, Azerbaijan. We examined the impact of radiation on erythrocyte nuclear pathologies, levels of cytochrome P-450, and serotonin-modulating anticonsolidation protein (SMAP) in the tissues of the hens from three settlements with different levels of background radiation. Higher levels of radiation resulted in increased nuclear pathologies, upregulation of tissue SMAP levels, and downregulation of cytochrome P-450. We also carried out controlled dosage studies on Wistar male rats, which showed significant upregulation of heat shock proteins with molecular mass 70 kDa (HSP70) in the bone marrow 3 and 5 h after SMAP intraperitoneal administration. Administration of SMAP to rats 3 h prior to γ-radiation exposure (8 Gy) provided significant protection to somatic cell nuclei. We conclude that SMAP can provide protection from the genotoxic effects of γ-radiation through upregulation of HSP70 or the transformation of chromatin into a condensed, more protective conformational state.

[Reparative mechanisms in retina and hypothalamus contributing to rhodopsin recovery in rabbit eyes with retinal dystrophy]. / Issledovanie reparativnykh mekhanizmov v setchatke i gipotalamuse, obespechivaiushchikh vosstanovlenie rodopsina v usloviiakh distrofii setchatki u krolikov.

PURPOSE: To study the molecular reparatory mechanisms of the retina and hypothalamus in the context of experimental dystrophy of receptor apparatus in rabbit retina. MATERIAL AND METHODS: Retinal dystrophy was induced in rabbit eyes by injecting monoiodacetic acid (MIA) intravenously. Indirect ELISA test was used to evaluate the levels of rhodopsin, heat shock protein 70 kDa (HSP70) in the retina, and serotonin-modulating anticonsolidation protein (SMAP) - that directly correlates with serotonin level - in the hypothalamus. RESULTS: The 1st series of studies showed that 12 days after the administration of MIA, rhodopsin in the retina was down-regulated by 27% (p<0.001), and heat shock protein 70 kDa (HSP70) was up-regulated by 47% (p<0.001), whereas in the hypothalamus up-regulation of SMAP by 22% (p<0.01) was observed. In the 2nd series, on the 22nd day after MIA administration, significant down-regulation (by 9.5 times) of HSP70 (p<0.001) was noticed in control rabbits, though intravitreal administration of SMAP on the 15th day after MIA administration led to sharp (23 times) up-regulation of HSP70 (p<0.001) in the retina 7 days later. In the 3rd series, 7 days after intravitreal administration of inactivated SMAP, the animals getting injections of MIA had noticeable down-regulation of rhodopsin (p<0.01) in the retina. In the 4th series, 7 days after intravitreal administration of polyclonal antibodies to SMAP in the rabbits that has had MIA injections, up-regulation of rhodopsin (p<0.01) and HSP70 (p<0.01) in the retina compared with levels in the control animals (MIA and non-immune γ-globulins) was observed. CONCLUSION: The results indicate the influence of the hypothalamic serotonergic system on HSP70 level in the receptor cells of the retina. The results of the 4th series of studies also give evidence for possible feedback from the retinal cells onto hypothalamus.

Does serotonin-modulating anticonsolidation protein (SMAP) influence the choice of turning direction in carps, Cyprinus carpio, in a T-maze?

Serotonin-modulating anticonsolidation protein (SMAP) can impair the formation of memory traces in mammals and fish. We have studied the influence of SMAP on behavioral lateralization of juvenile carps Cyprinus carpio in a T-maze without food reinforcement in three experimental groups (n = 8 each): (1) negative control (intact animals); (2) experimental group (fish injected ICV with SMAP; 2 µl, 1.2 mg ml(-1)) and (3) active control group (fish injected ICV with inactivated SMAP). The behavioral lateralization of carps was observed on the 1st, 2nd, 3rd and 6th days after the injections. In each observation session, a fish was placed five times in a start chamber of the T-maze. The direction of the turn upon leaving the start chamber, as well as the latency from the opening of start chamber flap to the fish's turn was registered. The number of right turns (of all five turns observed during the session) was a criterion of lateralization. It was found that carps have no inherent preference for turning left or right. The SMAP injection did not influence the choice of turning direction, but increases latency values insignificantly. The results are important for the correct interpretation and clarification of data reporting the role of SMAP in training and formation of spatial memory of fish in a maze.

[ENGAGEMENT OF SEROTONIN-MODULATING ANTICONSOLIDATION PROTEIN IN REGULATION OF EMBRYOGENESIS OF LYMNEAE STAGNALIS AND LEWIS SARCOMA IN HYBRID MICE Fl C57B2/6 X DBA].

The article concerns study of the effects of a novel serotonin-modulating anticonsolidation protein (SMAP) being in a linear relationship with serotonin level, on embryogenesis of Lymneae stagnalis and Lewis sarcoma in hybrid mice Fl C57B2/6 X DBA. Inhibition of embryogenesis of Lymneae stagnalis on the stage of four blastomers and late blastula, lack of changes on the stage of trochofora and acceleration of metamorphosis under the effects of SMAP in a dose-dependent manner was observed. Short-term retardation (during the first 10 days) of development of Lewis sarcoma in mice and survival of 25% of transferring animals under high doses of SMAP was revealed. Cytostatic activity for high doses of SMAP and their effects on the duration of single phases of the cell cycle is proposed.

[Effect of serotonin-modulated anticonsolidation protein on formation of long-term memory in carps Cyprinus carpio in the model of active avoidance learning].

Effect of serotonin-modulated anticonsolidation protein (SMAP) that has property of disturbing formation of memory trace in mammals and of learning and memory in teleost fish was studied in the model of active avoidance learning. The experiment was performed in three stages: (1) fry of carps Cyprinus carpio L. was injected intracerebrovenricularly with the SMAP protein at a dose of 0.3 µg/g; control individuals were administered with equal amount of the buffered saline for poikilothermic animals; (2) 24 h after the injection, fish were learnt during 8 sèances for 2 days the conditioned reflex of active avoidance; (3) 48 h after the learning the testing of the skill was performed. The administration of the protein was shown to lead to disturbance of reproduction of the skill in the fish: the latent time of the skill reproduction in experimental individuals exceeded that in control fish more than two times, while the number of individuals succeeding the task in the experimental group was non-significantly lower than in the control group. However, unlike mammals, injection of the SMAP protein in this model produced no effect on the process of learning in carps. Thus, there was first demonstrated the inhibiting effect of the SMAP protein whose concentration correlated positively with the content of the neurotransmitter serotonin in brain on consolidation of memory traces in teleost fish.

[Character of changes of the level of serotonin-modulating anticonsolidation protein and of cytochrome P-450 in tissues of the eastern alburnoid Alburnoides bipunctatus eichwaldi from rivers of Azerbaijan].

The paper deals with study by the method of solid-phase indirect immunoenzyme analysis of levels of the novel serotonin-modulated anticonsolidation protein (SMAP) that is directly correlated with serotonin level as well as of biomarker cytochrome P-450 in the liver, gills, and brain of the eastern alburnoid (Alburnoides bipunctatus eichwaldi) caught in the rivers Khudat, Akstafachai, Kura, and Araks flowing at the territory of Azerbaijan. There was revealed a marked downregulation of cytochrome P-450 and SMAP in the liver and gills of the fish caught in the Akstafachai River relatively to values in the fish from the Khudat River not contaminated with pollutants. In the liver and gills in the fish from the Kura and Araks rivers, a significant differently directed changes of the cytochrome P-450 and SMAP levels were observed: downregulations of the cytochrome P-450 versus an upregulation of SMAP. In the brain of the fish from the River Akstafachai there was observed some downregulation of cytochrome P-450, whereas in fish from the Kura and Araks rivers--a significant upregulation of the SMAP level. The obtained results are analyzed from standpoint of processes of adaptation and disadaptation of aquatic organisms to impact of pollutants.

[Inhibition of morphine intake by antibodies to serotonin-modulating anticonsolidation protein in model of self-administration in rats].

The article concerns study of effects of polyclonal antibodies to serotonin-modulating anticonsolidation protein (SMAP) being in direct dependence on serotonin level and providing intracellular transduction of serotonergic signal, on positive reinforcement effect of morphine in rats. The task was formed in Wistar male rats in the model of morphine self-administration as a result of pressing of one of two levers attached to the wall, joined to the pump delivering each time 100 µg of morphine directly into the vena jugularis. In the 1st series of studies brain cingulate cortex and hypothalamus were taken from the rats achieved stable level of morphine intake and SMAP level was measured with indirect immune-enzyme assay. It was shown that in the morphine-self-injected rats SMAP level in the cingulate cortex is significantly upregulated (p = 0.01), while in the hypothalamus it was left unchanged. In the 2nd series of studies the rats with stable level of morphine intake were administered intraperitoneally with anti-SMAP rabbit polyclonal antibodies (experimental group) or non-immune γ-globulins (control group). Soon after antibodies administration the animals of the experimental group demonstrated manifold decrease of morphine intake lasted for 8 days (p < 0.008), whereas it did not change in the controls. SMAP upregulation in the brain cingulate cortex in the rats with stable morphine intake, obviously, indicates to its engagement in positive reinforcement effect of morphine. Blockade of SMAP activity with anti-SMAP antibodies in the nerve cells induced sharp decrease of morphine intake due to disturbances of transduction through intracellular serotonin's signal channels.

[Adaptive increase of serotonergic system activity in tissues of half-migratory and migratory fish at increased water salinity].

The article deals with studies of the serotoninergic system activity in different tissues of half-migratory fish--the Caspian roach (Rutilus rutilus caspicus) and carpbream (Abramis brama orientalis)--and migratory fish--shemaya (Chalcalburnus chalcoides) caught in fresh and brackish waters, as well as in the common carp (Cyprinus carpio L.) tissues under effect of brackish water in model experiments. Using indirect solid-phase ELISA-test, the serotoninergic system activity was evaluated by measuring in the tissues of the studied fish the serotonin-modulated anticonsolidation protein (SMAP) which is in linear relationship with serotonin level. There was found a significant elevation of the SMAP levels in the brain of the Caspian roach, carpbream, shemaya, and the common carp under effect of increased water sainity. The revealed increase of the SMAP content in brains of the Caspian roach, carpbream, shemaya, and the common carp under action of increased water salinity reflects the corresponding elevated activity of the serotoninergic system and indicates involvement of adaptive readjustments in the animals' body.

[The role of serotonin-modulated anticonsolidation protein in memory formation in rats in a shuttle box].

Two series of experiments were carried out in Wistar male rats. In the first series, rats were trained to acquire conditioning in a shuttle box to 50% and 80% learning criteria. In the animals of the experimental group that achieved 50% learning criterion, a significant decrease in the levels of serotonin-modulated anticonsolidation protein (SMAP) (solid phase, indirect ELISA-test) was observed in the temporal cortex as compared to the animals of the active control group. In the animals of the experimental group that achieved 80% learning criterion, such a decrease was found in the occipital and temporal cortex. In the second series of the experiments, animals of the experimental group were injected with SMAP in saline at a concentration of 1.5 mg/ml in a volume of 10 microl through the cannula implanted into the left lateral ventricle of the brain. Control animals were administered with heating-inactivated SMAP in the same amount. The substances were injected to the animals under light ether anesthesia daily 40 min prior to learning sessions. Learning sessions were carried out in the shuttle box for several days to 50% learning criterion. The experimental rats achieved learning criterion within 7-8 days, whereas intact and control animals reached the same criterion within 4 days. Furthermore, the experimental group of animals differed in increased levels of fear, anxiety and aggression which did not decline throughout the whole learning period. The conclusion was made that SMAP participated in negative regulation of the memory trace formation.

[Participation of serotonin-modulated anticonsolidation protein in mediation of action of adverse factors on lipid peroxidation level in juvenile sturgeon tissues Acipenser güldenstädti persicus].

The work analyzes participation of the serotoninergic system in correction of adverse effects of benthic deposits contaminated with industrial wastes on the level of lipid peroxidation (LPO) in the fry of the sturgeon Acipenser güldenstädti persicus. In the first series of the study, two intramuscular injections to the fry of the sturgeon of serotonin-modulated anticonsolidation protein (SMAP) that is in the direct correlation with the serotonin level led to pronounced normalization in the animal tissues of the LPO level whose increase was due to keeping the sturgeons in the fresh water contaminated with benthic deposits. In the second series of the study, administration to the animals of polyclonal anti-SMAP antibodies led to a significant increase of the LPO level in the tissues, similar to effects of the benthic deposits. It is concluded that adverse factors can produce their negative prooxidative effects on the organism tissues through a decrease of activity of the serotoninergic system, whereas an artificial increase of the serotoninergic system activity promotes correction of the oxidative stress.

[Increase of mutation level in tissues of goby and fry of sturgeon under conditions of block by antibodies against serotonin-modulated anticonsolidation protein].

The work present data on studies of a decreased activity of serotoninergic system on the level of mutagenic changes (the micronuclear test) in the goby Neogobius fluviatilis and the fry if sturgeon Acipenser güldenstädti persicus. It has been shown that the long exposure of the animals to conditions of industrial and oil pollution leads to a significant decrease in their liver of the level of serotonin-modulated anticonsolidation protein (SMAP) correlating directly with the serotonin level as well as to sharp increase of the level of micronuclei in erythrocytes. The intramuscular administration of anti-SMAP polyclonal antibodies to the fry of the sturgeon produces a significant increase of the amount of micronuclei as compared with that in the animals injected with non-immune gamma-globulin. The obtained results allow concluding that the decrease of activity of the serotoninergic system is the mechanism that is triggered with adverse environmental factors and realizes mutagenic damages in the modified genetic apparatus.

[Antimutagenic activity of serotoninergic system and its mechanisms in Acipenser gueldenstaedti persicus and Carassius auratus].

The paper deals with antimutagenic body activity and its underlying mechanisms. The experiments carried out on the one-year old sturgeons (Acipenser gueldenstaedti persicus) and goldfish (Carassius auratus) have shown that intramuscular administration of serotoninmodulated anticonsolidation protein (SMAP) leads to a twofold decrease of erythrocyte mutagenic alterations (the micronuclear test, p < 0.01) caused by action of benthic deposits (0.8 ml/l, 3 days) polluted with industrial wastes. Exposure of goldfish in water contaminated with oil (500 mg/l, 3 days) led to a sharp rise of the content of the 70 kDa brain protein fraction (p < 0.001); these water-soluble proteins are assumed to belong to heat shock proteins (HSP). At the same time, in the brain of the studied animals there was observed a simultaneous increase of the SMAP content (p < 0.001). After 3 h, intracerebral SMAP administration to goldfish increased significantly the 90 kDa protein fraction content (p < 0.01), probably HSP90, in the electrophoretic profiles of the brain water-soluble proteins. Thus, the obtained results indicate that the body serotoninergic system has the antimutagenic activity providing protection of cells from action of harmful environmental factors by an enhancement of synthesis of proteins suggested to belong to HSP.

[Changes of the serotonergic system activity in fish tissues during an increase of water salinity].

Changes of environment parameters produce essential effects on the serotonergic system activity in the body tissues. This paper describes study of the serotonergic system activity in various tissues of half-migratory fish - the Caspian roach Rutilus rutilus caspicus and the bream Abramis brama orientalis - caught in the fresh and brackish water. Using the method of indirect solid-phase ELISA test, the serotonergic system activity was assessed by determining in the studied fish tissues of the amount of serotonin-modulating anticonsolidation protein (SMAP) that is in the linear correlation with serotonin level. A significant change of the SMAP level in the Caspian roach and bream brain was found under effect on increased water salinity. Analysis of the SMAP amount in the bream liver did not reveal any differences in the animals caught in the brackish and fresh water. At the same time, in the bream gill there was a small rise of the SMAP content in the fish caught in the brackish water. The revealed in this experiment elevation of the SMAP amount in the Caspian roach and bream brain reflecting a corresponding increase of the serotonergic system activity indicates involvement of adaptive readjustments in the animal body.

Decrease in activity of the serotoninergic system during mutagenesis.

We studied changes in the content of serotonin-modulated anticonsolidation protein in the liver of goldfishes and gobies caused by oil and industrial pollution. The concentration of serotonin-modulated anticonsolidation protein in fish liver increased after short-term exposure to oil-contaminated water (100 mg/liter), but decreased under long-term effect of industrial wastes. We hypothesize that serotonin plays a role in antimutagenic protection of the organism and maintains the differentiated state of mature cells.

Selective effects of antibodies to protein SMP-69 on the activity of defensive behavior command neurons in the common snail.

The effects of antibodies to the serotonin-modulated protein SMP-69 on the activity of defensive behavior command neurons LP11 and RP11 in semi-intact preparations from common snails were studied. Antibody to SMP-69 increased membrane excitability and facilitated neuron responses to chemical sensory stimulation by application of dilute quinine solution to the animal's head, these effects being seen at 1-1.5 h. The synaptic effects of the antibodies were specific, as they had no influence on responses induced by tactile stimulation of the head. The neuronal effects of antibody SMP-69 were similar to changes in the activity of cells LP11 and RP11 induced by serotonin and cAMP, and to changes seen when snails acquired nociceptive sensitization. It seems likely that a protein homologous to mammalian SMP-69 is involved in the mechanisms controlling excitability and long-term specific plasticity of the synaptic inputs to neurons LP11 and RP11 from chemoreceptors on the snail's head.

[Selective effect of the antibody to protein SMP-69 on activity of the defence behavior command neurons in grape snail]. / Izbiratel'noe vliianie antitel k belku SMP-69 na aktivnost' komandnykh neironov oboronitel'nogo povedeniia vinogradnykh ulitok.

Effects of antibody against serotonin-modulated protein SMP-69 on defence behavior command neurons L-RP11 were studied in semi-intact preparation of snail Helix lucorum. An increase in membrane excitability as well as selective facilitation of neural responses evoked with chemical sensory stimulation of the snail head (0.25-0.5% quinine solution) were determined 1-1.5 hours after antibody application to the neurons. The antibody did not change neural responses evoked with tactile stimulation of the snail head. These effects were similar to those found in L-RP11 neurons after serotonin or cAMP applications as well as after nociceptive sensitization of the snail. It was suggested that protein homologically related the SMP-69 in mammalians was involved in mechanisms of excitability as well as long-term specific plasticity regulation of L-RP11 neurons synaptic inputs from the head chemoreceptors in snail Helix lucorum.

Detection of protein with anticonsolidation properties in the rat brain.

Isolation and identification of rat brain protein regulated by serotonin-modulated protein SMP-69 is described. Intracerebral administration of anti-SMP-69 antibodies activates cerebral cortex cell genome and increases the content of electropheretic fraction 28. Intracerebral administration of isolated and purified protein to rats disturbed memory consolidation. It is concluded that nerve cells have a molecular switch regulating consolidation of memory traces.

[The effect of antibodies to the serotonin-modulating protein SMP-69 on passive avoidance behavior in rats]. / Vliianie antitel k serotonin-moduliruemomu belku SMP-69 na povedenie passivnogo izgebganiia u krys.

Previously we purified from rat brain the novel protein SMP-69 modulated by serotonin. Intracerebral injections of the antibodies to this protein produced an increase in exploratory activity of rats in the open-field test. In the present work we studied the influence of intracerebral injections of SMP-69 protein polyclonal antibodies on passive avoidance behavior of rats. Injections of antibodies 24 h prior to learning led to an increase in rats' stay in the dark compartment and decrease in time spent in the illuminated compartment in comparison to the control animals (injected with rabbit non-immune serum) during retest sessions conducted 48 h after learning. No significant difference was observed between the control and experimental rats in the retest sessions conducted 24 h after learning. Injections of antibodies 48 h after learning did not change the behavior of the experimental rats in retest sessions, conducted 24 h after the injections. The results suggest that the antibodies to SMP-69 affect the memory consolidation but do not influence the storage and retrieval.