The Effects of Repeated Administration of the Micellar Complex of Methylprednisolone on the Locomotor Activity of a Terrestrial Snails.

We studied the effects of repeated injections of methylprednisolone and its micellar complex with block-copolymer on locomotor activity of a terrestrial snail. It was shown that methylprednisolone solution injected into the hemolymph of the animal produced a direct effect on the muscle system of the animal as soon as 1 h after administration: it slowed down snail locomotion and reduced contractile activity of the foot muscles. The micellar complex of methylprednisolone with block-copolymer prevented this effect during the first 2 days of injection and negatively affected locomotion only in 2 days after injection, the decrease in locomotion in this case was not accompanied by a decrease in contractile activity of the foot muscle.

Changes in Membrane and Threshold Potentials of Command Neurons in Terrestrial Snail during Development of a Conditioned Situational Defensive Reflex.

Changes of the electrical characteristics of command neurons of defensive behavior caused by the development of a conditioned situational defensive reflex were studied experimentally under in vitro conditions on preparations of the nervous system of snails. After learning, the membrane and threshold potentials of command neurons LPa3 and RPa3 significantly decreased and excitability of the studied neurons increased.

Effects of NO Synthase Blocker L-NAME on Functional State of the Neuromotor System during Traumatic Disease of the Spinal Cord.

Functional state of the neuromotor system after administration of a nonspecific NO synthase blocker L-NAME was studied on the model of experimental contusion of the spinal cord. Electron paramagnetic resonance measurements of NO production in the damaged segment of the spinal cord were performed for estimation of the dynamics of intensity of NO production during traumatic disease of the spinal cord and selection of optimal period for L-NAME administration. The status of the neuromotor system was evaluated by stimulation electromyography. Treatment with L-NAME during the acute period of traumatic injury to the spinal cord sharply reduced the intensity of evoked motor responses and more pronounced increase in excitability of peripheral motor structures. The results suggest that NO system is a factor of regulation of the stress-induced and adaptive responses of the body at the early stage of spinal cord injury.

[The role of ATP-dependent potassium channels and nitric oxide system in the neuroprotective effect of preconditioning]. / Rol' ATF-chuvstvitel'nykh kalievykh kanalov i oksida azota v zashchitnom effekte prekonditsionirovaniya mozga.

AIM: To study a role of ATP-dependent potassium channels (K+ATP) in the neuroprotective effect of ischemic (IP) and pharmacological (PP) preconditioning and evaluate the dynamics of blood nitric oxide (NO) metabolites in cerebral ischemia. MATERIAL AND METHODS: A model of ischemic stroke induced by the electrocoagulation of a middle cerebral artery (MCA) branch was used in male rats (n=86). Glibenclamide, a selective inhibitor of ATP-sensitive K+ channels, and diazoxide, a potassium channel activator, were used. IP and PP were performed 24 h before MCA occlusion. Blood concentrations of NO, NO3- and NO2-were measured 5, 24 and 72 h after occlusion. RESULTS: IP decreased a lesion area by 37% (p<0/05) and the preliminary introduction ofglibenclamide levelled the effect of IP. A protective effect of PP was similar to that of IP. A decrease in oxygenated R-conformers of Hb-NO and a reverse increase in non-oxygenated T-conformers as well as NO3- и NO2-were noted 5h after MCA occlusion. In the first 24 h after MCA occlusion, contents of NO3- and NO2- returned to normal values. There were changes in the concentrations of Hb-NO complexes as well, with the predominance of R-conformers and minimal contents of T-conformers. Moreover, the correlations between K+ATP channel blockade and the decrease in serum NO3- and NO2 were found (p<0/03). CONCLUSION: The neuroprotective effect of preconditioning is caused by the activation of K+ATP channels. An analysis of NO metabolite concentrations in the blood of rats with IP suggests that Hb-NO complexes belonging to R-conformers deposit and carry NO in tissues releasing NO accumulated via R→T transfer in conditions of ischemia.

Effects of NO Donors and Inhibitors of NO Synthase and Guanylate Cyclase on the Acquisition of a Conditioned Defense Food Aversion Response in Edible Snails.

Experiments on edible snails revealed that NO synthase blockade with a nonspecific neuronal NO synthase inhibitor L-NAME before defense food aversion conditioning is followed by a decrease in the rate of learning. Exogenous NO donors, sodium nitroprusside and dinitrosyl iron complex, were shown to improve learning. Chronic administration of a specific soluble guanylate cyclase inhibitor ODQ also increased the rate of learning. Our results indicate that NO has a modulatory effect on the formation of a conditioned response in this type of learning.

Effect of NO synthase blockade on NO production in rat heart under conditions of hypokinesia.

Electron paramagnetic (EPR) spectroscopy study showed that 90-day hypokinesia in rats is accompanied by an increase in NO production in the heart. A nonselective NO synthase inhibitor L-NAME decreased the content of NO in the heart atria and ventricles of hypokinetic rats by 67-70%. A selective inhibitor of inducible NO synthase, aminoguanidine, also decreased the level of NO in the heart atria and ventricles of hypokinetic rats by 60-65%. Our results indicate that the increase in NO production during hypokinesia is associated with activation of NO synthases.

[Influence of hypokinezia of varying duration on the nitric oxide productions dynamics in rat's heart, spinal cord and liver].

By electron paramagnetic resonance method analyzed the production of nitric oxide (NO) in atrias and ventricles of the rat's heart, spinal cord and liver of rats after the contained in conditions of hypokinesia during 30, 60 and 90 days. It was discovered that the regime of hypokinesia leads to increase NO production in all the investigated tissues, and the largest increases were observed in 30-day hypokinesia.

[Aqueous and salt solutions of quinine of low concentrations: self-organization, physicochemical properties and actions on the electrical characteristics of neurons].

Self-organization, the physicochemical properties of aqueous and salt solutions of quinine and the effects of salt quinine solutions in a wide range of concentrations (1 x 10(-22) - 1 x 10(-3) M) on the electrical characteristics of the edible snail's identified neurons were studied. Similar non-monotonic concentration dependencies of physicochemical properties of aqueous and salt quinine solutions at low concentrations are obtained. This allows of predicting the occurrence of biological effects at low concentrations of quinine solutions. Intrinsic (within 5% of the interval) changes in membrane potential, the amplitude and duration of the neuron action potential under the influence of quinine salt solutions at concentrations of quinine of 1 x 10(-20), 1 x 10(-18), 1 x 10(-10) M are found. For these concentrations the extreme values of specific conductivity and pH are shown.

[Electron paramagnetic resonance study of nitric oxide production in rat heart during hypokinesia].

By the method of EPR spectroscopy we studied the intensity of nitric oxide (NO) production after modeling of hypokinesia in rats (limitation of moving activity) through analyses of the amount of NO-containing paramagnetic complexes in tissues of heart and liver. Experimental animals were kept during 60 day protracted hypokinesia. NO amount was assessed by the intensity of EPR spectra of complex (DEDTC)2-Fe(2+)-NO. It was established that after 60 day hypokinesia the 2-3 fold increase of NO quantity occured in heart and liver tissues. The application of a nonspecific inhibitor of NO-synthases, L-NAME, in suspended rats led to a decreased NO quantity up to the value more lower than in control. The obtained results show that the main contribution to the increase in the intensity of NO production during hypokinesia belongs to the fermentative pathway of NO production and the formation of NO by nitrite-reductase activity is not enhanced during hypokinesia.

Nitric oxide level in the rat tissues increases after 30-day hypokinesia: studies by electron paramagnetic resonance (EPR) spectroscopy.

Studies by EPR spectroscopy showed that 30-day exposure of rats to augmenting hypokinesia led to a 3-fold increase in nitric oxide (NO) production in the heart and 2-fold in the liver. These results indicated that long-term hypokinesia stimulated NO synthesis.

[Changes of the neuronal membrane excitability as cellular mechanisms of learning and memory].

In the presented review given literature and results of own studies of dynamics of electrical characteristics of neurons, which change are included in processes both an elaboration of learning, and retention of the long-term memory. Literary datas and our results allow to conclusion, that long-term retention of behavioural reactions during learning is accompanied not only by changing efficiency of synaptic transmission, as well as increasing of excitability of command neurons of the defensive reflex. This means, that in the process of learning are involved long-term changes of the characteristics a membrane of certain elements of neuronal network, dependent from the metabolism of the cells. see text). Thou phenomena possible mark as cellular (electrophysiological) correlates of long-term plastic modifications of the behaviour. The analyses of having results demonstrates an important role of membrane characteristics of neurons (their excitability) and parameters an synaptic transmission not only in initial stage of learning, as well as in long-term modifications of the behaviour (long-term memory).

[The effect of serotonin precursor 5-hydroxytryptophan and neurotoxic analogue 5,7-dihydroxytryptamine on defensive conditioning in a snail].

This study is devoted to investigation of the influence of precursor of serotonin 5-hydroxytryptophane (5-HTP) and neurotoxic analogue 5,7-dihydroxytryptamine (5,7-DHT) on defensive conditioning and electrical characteristics of command neurones of defensive behaviour after learning. Snails injected with 5-HTP learned faster as compared to control group injected with physiological solution. After the 5,7-DHT injection, snails failed to form the conditioned reflex. Injection of 5-HTP after the preliminary injection of neurotoxin 5,7-DHT restored the capability of snails for learning. Injections of 5-HTP prevented the effect of 5,7-DHT at the behavioural level, but not at the level of electrical characteristics of the command neurones.

Electrophysiological studies of the effects of chronic administration of caffeine on the formation of a conditioned defensive reflex in the common snail.

Chronic administration of caffeine to common snails increased the rate of formation of a conditioned defensive reflex. When daily caffeine injections were given immediately after the training procedure, the conditioned defensive reflex was acquired more quickly than when caffeine injections were given before the training procedure started. Chronic caffeine administration to both trained and untrained snails led to depolarization changes in the membrane potential and reductions in the threshold potential of defensive behavior command neurons in common snails. Comparative studies showed that addition of caffeine to the solution bathing the mollusk nervous system resulted in decreases in the threshold potential of command neurons in both intact and trained snails; there was, however, no change in the membrane resting potential.

Serotonin precursor 5-hydroxytryptophan disturbs the protective effect of low doses of antibodies to S100B protein during the formation of long-term sensitization.

We studied the effect of ultralow doses of antibodies to calcium-binding protein S-100B and 5-hydroxytryptophan, a metabolic precursor of serotonin, on the formation of long-term sensitization as a neurobiological model of anxiety and depression. Daily administration of antibodies to S-100B to edible snail before the formation of long-term sensitization prevents its development. 5-Hydroxytryptophan administered before the formation of long-term sensitization abolished the protective effect of antibodies to S-100B protein.

Changes in electrical properties of command neurons during protective effect of low doses of antibodies to S100 protein on the development of long-term sensitization in Helix lucorum.

We studied the effect of antibodies to Ca(2+)-binding protein S100 in a dilution of 10(-12) (LAT-S100) on the development of long-term sensitization in Helix lucorum, a neurobiological model of anxious and depressive states. After administration of LAT-S100 preventing the development of long-term sensitization before training, the membrane and threshold potentials in command neurons regulating defense behavior decreased less markedly than during long-term sensitization. It is assumed that the "protective" effect is associated with mechanisms of long-term potential maintenance and changes in intra- and extracellular balance of Ca(2+)-binding protein S100.

Effect of neuroleptic haloperidol, L-DOPA precursor dopamine, and neurotoxic dopamine analog 6-OHDA on acquisition of conditioned defensive reflex in edible snail.

The effect of neuroleptic haloperidol, a dopamine precursor L-DOPA, and dopamine analog 6-OHDA on the development of conditioned defensive reflex was studied in edible snails. Injection of L-DOPA to intact snails and to snails pretreated with 6-OHDA 2 h before learning session decelerated acquisition of the conditioned reflex and exerted a toxic action. In contrast, injection of 6-OHDA or haloperidol did not affect acquisition of the conditioned reflex.

Effects of preliminary administration of haloperidol in low doses on the effects of haloperidol on behavioral reactions and command neuron membrane potential in edible snail.

We studied the effects of preliminary administration of haloperidol in low doses on changes in motor activity of edible snail and in electrical properties of defensive behavior command neurons induced by chronic administration of haloperidol. The rate of locomotion decreased after injections of haloperidol preparations (C6, C12, C30, C200 and a mixture C12+C30+C200) for 3 days. Similar changes were observed after 3 days of haloperidol administration. Haloperidol preparations in low doses produced a modulating effect on the decrease in locomotion rate and hyperpolarization of command neurons in edible snails caused by chronic exposure to haloperidol: the decrease in locomotion rate caused by chronic haloperidol treatment was prevented by preliminary injection of haloperidol in low doses C6, C12 and C30; the depolarizing shift of command neuron membrane potential was also abolished after consecutive injection of the same haloperidol preparations C6, C12 and C30.

Electron paramagnetic resonance study of nitric oxide production during 5-HT2 receptor blockade in the blood, heart, and liver of rats with myocardial infarction.

The effect of serotonin 5-HT2 receptor blockade on nitric oxide production in rats with myocardial infarction was evaluated by the method of electron paramagnetic resonance. The spectra were recorded in samples of the heart tissue (left and right chambers), liver, and blood. 5-HT2 receptor blockade during myocardial infarction was followed by a decrease in the total content of nitrosyl complexes in the spin trap and R and T conformers of Hb-NO. The percent of T conformers increased in the remaining complexes of Hb-NO. These changes were observed in the heart and, particularly, in the blood. The amount of spin-trap complexes was lower in the liver. Hence, nitric oxide molecules were primarily associated with the spin trap in liver tissue. The decrease in the number of Hb-NO complexes in the blood probably reflects the decrease in the severity of hypoxia due to myocardial infarction. A correlation was found between these changes and physiological state of rats. 5-HT2 receptor antagonist improved general state of rats with infarction and increased survival rate.

[Electrophysiological study of effects of chronic injection of caffeine on defensive reflex conditioning in grape snail].

It was found that chronic injection of caffeine to grape snail increases a velocity of elaboration of conditioned defensive reflex. It was shown that after daily injection of caffeine immediately after procedure of learning the conditioned defensive reflex elaborated faster than daily injection before procedure of learning. It has been shown, that chronic injection of caffeine both in naive as well as learned snails led to depolarizing shift of membrane potential and to decrease of threshold potential of command neurons of the defensive behaviour of grape snails. It was also found that addition of caffeine in bath solution led to decrease of threshold of generation of action potential of command neurons both in intact and learned snails. The resting membrane potential of command neurons was not changed.

Investigation of changes in NO content during long-term sensitization in edible snail using EPR-spectroscopy: effects of antibodies to calcium-binding protein S-100.

EPR-spectroscopy experiments (electron paramagnetic resonance) demonstrated a decrease in NO production in the nervous system and heart of edible snail Helix lucorum after formation of long-term sensitization, a neurobiological model of anxiety and depression. The protective effect of antibodies to Ca(2+)-binding protein S-100 in dilution of 10(-12) on the formation of long-term sensitization was accompanied by partial recovery of NO synthesis in the nervous system and heart. These findings indicate that the imbalance in Ca(2+)-binding protein S-100 can lead to inhibition or modulation of some processes during plastic reorganization in the body and especially during pathological processes.