Serotonin-modulating Anticonsolidation Protein Research Articles

Impact of dihydropyrimidinase-related protein 2 in memory formation on rats and its possible role in neuronal back remodeling

The article concerns the problem of molecular mechanisms of memory formation. In this study the effects of polyclonal antibodies to serotonin-modulating anticonsolidation protein (SMAP) complex and its component dihydropyrimidinase-related protein 2 (DRP2) have been analyzed. Intra-cerebral administration of polyclonal anti-SMAP antibody significantly enhanced elaboration and strengthened memory formation in two complex behavioral conditioned models. At the same time, intra-cerebral administration of anti-SMAP antibody resulted in an increase of the content of nerve growth factor (NGF) in the water-soluble fraction of the hippocampus while intra-cerebral administration of anti-DRP2 antibody caused a decrease in the content of β-III tubulin (a marker of differentiated neurons) in the hippocampus and in the left parietal cortex of untrained rats. The obtained results indicate that DRP2 might participate in regulation of the processes of back remodeling of mature nerve cells of adult organisms, occurring during training of rats in the behavioral paradigm used in this study under the effects of anti-SMAP and anti-DRP2 antibodies. Conclusion is made that back remodeling (dedifferentiation) of mature nerve cells, apparently, is engaged in memory formation.

The changes of serotonin-modulating anticonsolidation protein and dihydropyrimidinase-related protein 2 in the amygdala and blood of depressive rats

Nowadays, depression is referred to psychiatric disease and is widely spread throughout the world. The pathogenesis of depression in humans is characterized by significant disturbances of serotonin turnover and its downregulation in brain structures. The objective of the present study was the definition of the levels of serotonin-modulating anticonsolidation protein (SMAP) and dihydropyrimidinase-related protein 2 (DRP2) in the brain amygdala, platelets, and to anti-SMAP natural autoantibodies in the depression Wistar rats of both sexes. A depression state in the rats was formed in the dominant model and its successful formation was confirmed in the forced swimming test by the decreased timeframe of active swimming. Through the application of an indirect ELISA test in the depressive male and female rats, a downregulation of SMAP and DRP2 in the amygdala, an upregulation of SMAP in the platelets of depressive male rats, a noticeable downregulation of DRP2 in the platelets of depressive female rats, and a sharp downregulation of anti-SMAP natural autoantibodies in the serum of depressive male rats were observed.

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.

Activation of the Serotonergic System Promotes Survival of Different Animal Species Exposed to Bacterial and Chemical Toxins

We analyzed the role of serotonergic system in the protection of an organism of different animal species (mice, fish) from lethal doses of bacterial and chemical toxins. In the first series of experiments, peritonitis was induced in mice by i.p. administration of the pathogenic Escherichia coli strain, and the following i.m. administration of the serotonin-modulated anticonsolidation protein (SMAP) significantly increased their survival rate (75 vs. 30%, p < 0.05). In the second series, Western blot analysis showed that i.m. administration of SMAP to mice led after 5 h to a sharp upregulation of the 70 kDa heat shock protein (HSP70) in the liver. In the third series, carried out on the common carp Cyprinus carpio L., i.m. administration of SMAP prior to putting the fish into water containing a high concentration of a systemic insecticide Actara led to the survival of all individuals in the experimental group over the period of 5 days, whereas in the control (SMAP-free) group the survival rate over the same period was only 35% (p < 0.001). In the fourth series, administration of anti-SMAP polyclonal antibodies to C. carpio caused death of 75% of individuals that were put for 5 days into the water containing the sublethal Actara dose, whereas in the intact and control groups all individuals survived under the same conditions (p < 0.001).

The role of neuro-specific dihydropyrimidinase-related protein 2 (dpyl2) in spatial memory formation in teleosts

This article presents the results of an experiment on the influence of serotonin-modulating anticonsolidation protein (SMAP) on the spatial memory formation of juvenile goldfish Carassius auratus (L.) in a maze with food reinforcement. Three experimental fish groups were formed: (1) intact animals, (2) experimental group (fish injected ICV with SMAP in 24 h before the beginning of training; 2 μl, 1.5 mg·ml–1), (3) active control group (fish injected ICV with inactivated SMAP). Goldfishes of the experimental group demonstrated the lowest capability for spatial recognition: the maximum level of performance of the task was on 4th day of the training – 38%, while the values of this index in fishes of the control and intact groups were 70% and 63% respectively. In general, throughout the period of the training the average value of task performance was 16% in the SMAP-injected fish (in the control and intact groups – 42% and 53%, respectively). By using Ds-Na-polyacrylamide gel electrophoresis SMAP compositeon has been revealed. It was found that it consists of 10–12 protein components, among which four proteins dominated. They were identified by mass spectrometry MALDI-TOF: spectrin, dihydropyrimidinase-related protein 2 (DPYL2), tubulin and actin. It has been suggested that the most likely candidate responsible for the negative effects of SMAP on fish memory formation is DPYL2. It was hypothesized that anticonsolidation effect of SMAP is caused by the effect of DPYL2 which blocks the growth of axons or its cytostatic activity which leads to disorders in formation of new neurons in the brain as a result of learning.

Reparative mechanisms in retina and hypothalamus contributing to rhodopsin recovery in rabbit eyes with retinal dystrophy

To study the molecular reparatory mechanisms of the retina and hypothalamus in the context of experimental dystrophy of receptor apparatus in rabbit retina. 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. 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. 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.

Role of the brain in adaptation of the common and golden carps to adverse factors of chemical origin

Using an indirect solid-phase immunoassay (ELISA test), cytochrome P-450, serotoninmodulated anticonsolidation protein (SMAP) and 70-kDa heat shock protein (HSP70) levels were analyzed in the brain and liver of the common carp Cyprinus carpio L. and golden carp Carassius auratus caught in reservoirs of the Arpachay river (clean zone) and Araks river (polluted zone) in Nakhchivan Autonomous Republic of Azerbaijan. Upregulation of cytochrome P-450 (p < 0.05) and SMAP (p < 0.01) in the common carp brain and downregulation of cytochrome P-450 (p < 0.05) and HSP70 (p < 0.05) in the golden carp liver were detected in fish caught in the Araks river reservoir as compared to levels of the same substances in fish from the Arpachay river reservoir. It is concluded that the brain plays a regulatory role in adaptation of animals to adverse environmental conditions and in protection of tissues from negative effects of pollutants.

The preconditioning phenomenon and its mechanisms in the common carp as affected by Actara insecticide

The effect of Actara insecticide on the common carp (Cyprinus carpio Linnaeus) was studied, and its sublethal concentration (LC50) for this species was estimated. Exposure to Actara at a concentration of 400 mg/L in freshwater was shown to kill all the common carp individuals within 24 h, while 24-h preconditioning at an Actara concentration of 100 mg/L promoted survival of animals when they were subsequently exposed to the insecticide at a concentration of 400 mg/L for 5 days. Estimates of serotonin-modulated anticonsolidation protein (SMAP) in the common carp brain and liver by enzyme-linked immunosorbent assay (ELISA) following exposure to Actara at a concentration of 100 mg/L for 24 h showed its increased level in both organs (p < 0.001), particularly manifest in the liver. It is concluded that preconditioning elevates SMAP tissue levels and promotes thereby protection of the organism against damaging effects of lethal Actara concentrations.

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; 2μl, 1.2mgml(-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 6thdays 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.

Changes of serotonin-modulating protein and cytochrome P450 in tissues of eastern bystryanka (Alburnoides bipunctatus eichwaldii (Filippi, 1863)) and Caucasian river gobies (Neogobius constructor (Nordmann, 1840))

This article concerns studies of changes in biomarker P450 and the novel serotonin-modulating anticonsolidation protein (SMAP) in the brain, liver, and gills of eastern bystryanka (Alburnoides bipunctatus eichwaldii (Filippi, 1863)) and Caucasian river gobies (Neogobius constructor (Nordmann, 1840)) caught in rivers of Azerbaijan polluted with industrial wastes. The levels of both proteins in the fish tissues are evaluated with an indirect immune-enzyme assay. It is shown that both indexes, especially SMAP, have different patterns of changes in the tissues of bystryanka and gobies, which may be related to different ecological peculiarities of these species.

Character of changes in serotonin-modulating anticonsolidation protein and cytochrome P-450 in tissues of Alburnoides bipunctatus eichwaldi caught in the rivers of Azerbaijan Republic

The work was designed to investigate the levels of a novel serotonin-modulating anticonsolidation protein (SMAP), which activity directly depends on serotonin level, and biomarker cytochrome P-450 in the liver, gills and brain of Alburnoides bipunctatus eichwaldi caught in Khudat, Agstafachay, Kura and Araz rivers running over the territory of Azerbaijan. Noticeable downregulation of cytochrome P-450 and SMAP was observed in the liver and gills of fish caught in Agstafachay river relatively to the values obtained for the fish from unpolluted Khudat river. The levels of cytochrome P-450 and SMAP in liver of fish from Kura and Araz rivers exhibited significant opposite changes: downregulation of cytochrome P-450 level vs. upregulation of SMAP level. In the brain of fish from Agstafachay river the cytochrome P-450 was found to be somewhat downregulated, whereas in the brain of fish from Kura and Araz rivers SMAP was significantly upregulated. The results obtained are analyzed from the standpoint of adaptation and disadaptation processes of water organisms to the effects of pollutants.

Effect of serotonin-modulated anticonsolidation protein on formation of long-term memory in the learning model of active avoidance in the carp (Cyprinus carpio)

Effects of serotonin-modulated anticonsolidation protein (SMAP) that has property to impair formation of memory trace in mammals and of learning and memory in teleost fish was studied in the learning model of active avoidance. The experiment was performed in three stages: (1) fry of carp Cyprinus carpio L. was injected intracerebroventricularly with the SMAP 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 injection, fish were learned the conditioned reflex of active avoidance; (3) 48 h after the learning the testing of the skill was perfomed. The administration of the protein was shown to lead to impairment 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 expreimental group was insignificantly lower than in the control group. However, unlike mammals, injection of the SMAP in this model produced no effect on the process of learning in carps. Thus, there was first demonstrated the inhibiting effect of the SMAP whose concenration correlated positively with the content of the neurotransmitter serotonin in brain on consolidation of memory traces in teleost fish.

Adaptive increase of level of serotonin-modulated anticonsolidational protein in tissues of semimigratory and migratory fish at increase of water salinity

The article deals with studies of serotonergic system activity in different tissues of semimigratory fish—the Caspian roach (Rutilus rutilus caspicus) and the carpbream (Abramis brama orientalis)—and of 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 the indirect solid-phase ELISA-test, the serotonergic 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 the serotonin level. There was found a significant elevation of SMAP levels in brains of the Caspian roach, carpbream, shemaya, and the common carp under action of increased water salinity. A revealed increase of the SMAP content in brain 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.

ROLE OF DOWNREGULATION OF SEROTONERGIC SYSTEM IN CHANGING EXCITABILITY OF BRAIN STRUCTURES AND DEVELOPMENT OF EPILEPTIC SEIZURES IN RABBITS

The goal of the study was analysis of the role of novel serotonin-modulating anticonsolidation protein (SMAP; Mekhtiev, 2000) being in linear relationship with serotonin level and, perhaps, realizing its functions on subcellular level, in formation of seizure activity in rabbits. To achieve this goal SMAP blockade in the brain structures was realized with rabbit anti-SMAP polyclonal antibodies or autoantibodies produced in the organism of experimental animals as a result of immunization with SMAP. The results indicate to an important role of maintaining optimal level of serotonergic system’s activity in prevention of development of seizure activity as well as that down regulation of this system in the organism might timely inform about possible forthcoming advent of epilepsy fits.

Studies of the Role of Serotonin-Modulating Anticonsolidation Protein in Memory Formation in Rats in a Shuttle Box

Two series of experiments were performed on male Wistar rats. In the first series, rats were trained to a conditioned reflex skill in a shuttle box to the 50% and 80% learning criteria. A significant reduction in the level of serotonin-modulated anticonsolidation protein (SMAP) was observed in the parietal area of the cortex in animals of the experimental group reaching the 50% learning criterion and in the occipital and parietal areas of the cortex in animals of the experimental group reaching the 80% learning criterion, as compared with animals of the active control group. In the second series of experiments, animals of the experimental group received daily serotonin-modulated anticonsolidation protein at a concentration of 1.5 mg/ml in physiological saline (10 μl) via a cannula implanted into the left lateral ventricle of the brain 40 min before training sessions. Control animals were given the same quantity of heat-inactivated protein. Training was performed in a shuttle box for a period of several days to the 50% learning criterion. These experiments showed that rats of the experimental group reached the learning criterion in seven days, while intact and control animals reached this level in four days. In addition, animals of the experimental group differed in having increased levels of fear, anxiety, and aggressivity, which did not decrease throughout the training period. These results led to the conclusion that serotonin-modulated anticonsolidation protein is involved in the negative regulation of memory trace formation in nerve cells.

SUPPRESSION OF EPILEPTIC ACTIVITY IN RABBITS THROUGH SEROTONIN-MODULATING ANTICONSOLIDATION PROTEIN

The article concerns molecular basis of epileptic seizures development and elaboration of novel ways of their suppression through upregulation of serotonergic system activity on subcellular level by administration of serotonin-modulating anticonsolidation protein (SMAP). This protein had been purified earlier from the rat brains, possesses with Mr 186 kDa and is being in linear relationship with serotonin. Epileptic seizures were formed in Shinshilla rabbits by microapplication of 300 units of sodium penicillin solution into the brain amygdala. The evoked potentials (EP) to light flashes were fixed in the visual cortex, colliculus superior and retina. The epileptic seizures reached their peak for 50 min and lasted for 3 h. During their development, strengthening of the EP in the form of increasing their positive components, advent of interictal spikes and visually myoclonical seizures was observed. Administration of SMAP (3.5 ml, 1.5 mg/ml, i.m.) to rabbits 30 min after induction of epileptic seizures led to their suppression and elimination of their all manifestations including interictal spikes. The conclusion is done that upregulation of SMAP leads to suppression of epileptic seizures.

A rise of mutation level in tissues of monkey goby and sturgeon fry under conditions of block by antibodies to the serotonin-modulated anticonsolidation protein

The work presents data on studies of effect of a decrease in the serotoninergic system activity on the level of mutagenic changes (the micronuclear test) in the goby Neogobius fluviatilis and the fry of sturgeon Acipenser guldenstadti 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 a sharp rise of the level of micronuclear in erythrocytes. The intramuscular administration of anti-SMAP polyclonal antibodies to the sturgeon fry produces a significant increase of the amount of micronuclear as compared with that in the animals injected with non-immune γ-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.

Participation of serotoninmodulated anticonsolidation protein in mediation of action of adverse factors on lipid peroxidation level in tissues of sturgeon fry 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.

Antimutagenic activity of serotoninergic system and underlying mechanisms in fry of sturgeon and goldfish

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 serotonin-modulated 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 crude 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.