V. V. Andrianov

Modulation of defensive reflex conditioning in snails by serotonin

Highlights Daily injection of serotonin before a training session accelerated defensive reflex conditioning in snails. Daily injection of 5-hydroxytryptophan before a training session in snails with a deficiency of serotonin induced by the “neurotoxic” analog of serotonin 5,7-dihydroxytryptamine, restored the ability of snails to learn. After injection of the “neurotoxic” analogs of serotonin 5,6- and 5,7-dihydroxytryptamine as well as serotonin, depolarization of the membrane and decrease of the threshold potential of premotor interneurons was observed. We studied the role of serotonin in the mechanisms of learning in terrestrial snails. To produce a serotonin deficit, the “neurotoxic” analogs of serotonin, 5,6- or 5,7-dihydroxytryptamine (5,6/5,7-DHT) were used. Injection of 5,6/5,7-DHT was found to disrupt defensive reflex conditioning. Within 2 weeks of neurotoxin application, the ability to learn had recovered. Daily injection of serotonin before a training session accelerated defensive reflex conditioning and daily injections of 5-HTP in snails with a deficiency of serotonin induced by 5,7-DHT restored the snails ability to learn. We discovered that injections of the neurotoxins 5,6/5,7-DHT as well as serotonin, caused a decrease in the resting and threshold potentials of the premotor interneurons LPa3 and RPa3.

EPR study of nitric oxide production in rat tissues under hypokinesia

EPR spectroscopy was used to study the intensity of nitric oxide (NO) production upon modeling 60-day progressive hypokinesia (restriction of motor activity) in rats and estimating the content of (DETC)2-Fe2+-NO complexes in heart and liver tissues. In 30 days of hypokinesia, there was a 2–3-fold increase in tissue NO. Administration of a nonspecific inhibitor of NO synthases, L-NAME, to hypokinetic rats prior to measurement decreased their NO level even below the untreated control. Our results show that the intensified NO production in hypokinesia is mainly due to NO synthases, rather than to the nitrite reductase pathway.

Locomotor responses and neuron excitability in conditions of haloperidol blockade of dopamine in invertebrates and vertebrates.

Levels of movement activity were used to identify two groups of rats: those with high- and low-activity levels. Blockade of dopamine receptors with haloperidol led to suppression of locomotor activity in both groups of rats; in common snails, haloperidol decreased the rate of locomotion. The excitability of spinal centers in rats decreased 5 min after single i.v. injections, with gradual recovery seen by 30 min. Chronic administration of haloperidol suppressed post-tetanic potentiation of the H response in the gastrocnemius muscle of spinal rats. Prolonged use of haloperidol induced significant hyperpolarization of the membrane potential of command neurons in common snails and increased the action potential generation threshold. Selective pharmacological exclusion of the brain dopamine system was found to lead to decreases in the excitability of defined neurons in snails and the spinal motor centers in rats, also producing impairments in locomotor responses in these animals.

The electrical characteristics of command and motor neurons during acquisition of a conditioned defensive reflex and formation of long-term sensitization in snails

The mechanisms of the acquisition of a conditioned defensive reflex and formation of long-term sensitization were studied at the level of an analysis of electrical characteristics of defensive behavior command neurons and motor neurons which open and close the pneumostoma. Significant decreases in membrane and threshold potentials were seen in command neurons in response to acquisition of the conditioned defensive reflex and formation of long-term sensitization, along with further decreases in these parameters in snails which were trained after long-term sensitization. Changes, in the critical level of depolarization and amplitude of action potentials were insignificant. The changes observed here provide evidence for an increase in the membrane excitability of command neurons. Similar changes were not seen in motor neurons.

ESR study of the nitric oxide production in tissues of animals under an external influence on the functioning of the cardiovascular and nervous systems

Electron spin resonance (ESR) of the ternary (DETC)2-Fe2+-NO complex has been applied to determine the nitric oxide production in tissues of rats and snails. A preliminary ESR study of the NO content in tissues of rats before and after artificially induced acute myocardial infarct was performed. The analysis of the obtained results shows that the nitric oxide production during the first hour after the moment of inducing myocardial infarct decreases. It is also demonstrated that ESR may be useful in the study of the influence of the long-term sensitization of snails on the nitric oxide production in their body. The changes in the NO production after the external influences in both cases are discussed.

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 in nitric oxide in heart of intact and sympathectomized rats of different age

Nitric oxide production in heart tissues of rats of different age in the norm and after pharmacological sympathectomy was studied by electron spin resonance spin-trapping. Rats at the age of 14, 21, 70, and 100 days were used in the experiment. The concentration of nitric oxide produced in rat heart tissues proved to considerably decrease during ontogeny. Pharmacological sympathectomy notably decreased nitric oxide production in the heart in 14-and 21-day-old rats: the nitric oxide concentration in the spin trap as well as the level of R and T conformers of hemoglobin nitrosyl complexes decreased. In 70-day-old rats, pharmacological sympathectomy had no notable effect on the level of nitric oxide-containing paramagnetic complexes.

[The role of ATP-dependent potassium channels and nitric oxide system in the neuroprotective effect of preconditioning].

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.

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.

Study of effects of antibody to protein S100 on ionic channels of input and output currents of identified neurons of the snail Helix lucorum

In the course of electrophysiological experiments, two types of the neurons of the edible snail Helix lucorum were detected, which responded by different way to application of antibodies to the neuron-specific calcium-binding S 100 protein (AS1000). Under effect of AS100, frequency of the action potential (AP) generation in the spontaneously active V1, V3, V17, and RPa6 cells decreased, whereas in V4 and V6 cells increased. On addition of quinine solution the AP generation frequency of these neurons decreased more than twice, while the AP duration (tS) rose 6 times. The combined action of AS100 and quinine did not change statistically significantly the AP generation frequency, membrane potential (MP) and AP generation threshold (APt), as compared with the effect of AS100 in saline. The value of the AP duration (tS) increased 1.6 times, which was less pronounced as compared with the quinine action in saline. This means that AS100 prevents an increase of the AP duration after the quinine application (block of the Ca-depended K-channels). The main AS100 effect at the level of the ionic currents is shown to consist in a decrease of the maximal value of the input current, on average, by 20%, while of the output current, on average, by 12%.