Oxidative Damage of Proteins Precedes Loss of Cholinergic Phenotype in the Septal Neurons of Olfactory Bulbectomized Mice.

BACKGROUND: The development of cholinergic deficit is considered an early sign of a number of pathological conditions, including Alzheimer's disease. Cholinergic dysfunction underlies cognitive decline associated with both normal aging and Alzheimer's disease. OBJECTIVE: Here, we studied a possible mechanism of functional impairment of cholinergic neurons using an olfactory bulbectomy model. METHODS: Male mice were subjected to olfactory bulbectomy or sham surgery. Three weeks after that they were trained in Morris water maze and then euthanized one month after surgery. The cholinergic indices as well as the indices of oxidative stress were studied using immunohistochemistry, western blot and ELISA. Gene expression was studied using RT-qPCR. RESULTS: The experimental treatment was followed by impaired learning of a standard spatial task in a water maze. This was associated with a decrease in the number of cells containing choline acetyltransferase (ChAT), in relation to total number of neurons in the medial septum and lower ChAT enzymatic activity in the hippocampus. However, the levels of mRNAs of ChAT, vesicular ACh transporter and acetylcholine esterase remained unchanged in bulbectomized mice compared to sham-operated animals. These alterations were preceded by the accumulation of protein-bound carbonyls, indicating oxidative damage of proteins, whereas oxidative damage of nucleic acids was not detected. CONCLUSION: We assume that in olfactory bulbectomy model, oxidative damage of proteins may cause cholinergic dysfunction rather than irreversible neuronal damage. These data indicate that cholinergic neurons of the basal forebrain are very sensitive to oxidative stress, which may be responsible for the appearance of early cognitive decline in Alzheimer's disease.

[Structure of Response and Dynamics of Preferred Orientation in Cat's V1 Neurons].

Preferred orientation of V1 neurons was changed during the respons time course. In order to reveal mechanisms of such changes, spike trains of single neurons to flashed bars of different orientations were studied by means of principal component and frequency-domain analysis of the spike density functions. There was found out that: 1. contribution of each of the components was dependent on the stimulus orientation. 2. the principal components of V1 neurons responses contain different kinds of transient and sustained parts and rhythmic oscillations in the theta/alpha and beta-bands. 3. There was no correlation between transient parts as well as between rhythmic oscillations within the same frequency band in different components. Taken together, the data obtained allow us to suggest that during generation of response there is a shift of dominance of the components with different orientations which determines the dynamics of the preferred orientations of V1 neurons.

[The Effects of Chronic Combined Stress: Changes in Behavior of Rats with Various Responses to Novelty].

We studied the effect of chronic combined stress (model of experimental neurosis) on behavior of rats with different basal strategies of behavior in novelty conditions. Chronic stress resulted in decreases in the body weight and testosterone contents in the blood and neocortex in all animals. Animals with initially low orient- ing-exploratory response in the "open field" test did not exhibit substantial alterations of behavior during repeated testing in this test of the "dark-light chamber" test; however, the depression-like behavior was more expressed in the second forced swim test. Chronic combined stress did not significantly affect the behavior of this group of rats. Animals with initially high orienting-exploratory response in the "open field" test exhibited decreased locomotor and exploratory activity in the repeated "open field" tests. The decreases in the locomotor and exploratory activity were substantially less expressed in the repeated tests in these rats after chronic combined stress. The indices of depression-like behavior increased one month after the end of exposure to chronic combined stress. Our data demonstrate that different responses to novelty in the "open field" test do not allow predict with reasonable certainty the development of depression-like behavior after exposure to chronic combined stress.

[Effect of a Single Injection of Brain Derived Neurotrophic Factor into Midline Ventral Tegmental Area on Morphine Reinforcing Properties].

Opiate reinforcement is considered as a stimulus inducing addiction, however underlying neurobiological mechanisms remain obscure. According to the literature, BDNF (brain-derived neurotrophic factor) in the lateral ventral tegmental area (VTA) could modulate morphine reinforcement, but the role of BDNF in the midline VTA has not been studied yet. The aim of the study was to investigate the effect of a single intra-mid- line VTA injection on the acquisition and expression of morphine conditioned place preference (CPP). CPP procedure was composed of eight conditioning sessions (one session per day): morphine (i.p., 10 mg/kg) and saline injections were paired to the compartments and counterbalanced.Recombinant human BDNF (0.75 ug) or phosphate-buffered saline (PBS) as a vehicle were injected once into the midline VTA one day before or after conditioning. According to the CPP test rats spent more time in the morphine-paired compartments a scompared to the saline-paired compartments (p < 0.05). After a single BDNF injection into the midline VTA be- fore conditioning, but not after conditioning, differences in time spent in morphine and saline-paired compartments did not reach significance (p > 0.05). Thus, taking into account limitations of the results, we sug- gest that BDNF in the midline VTA may block morphine reinforcement.

[Comparison of its and orientation tuning dynamics in neurons located in different functional domains of cat's primary visual cortex].

Orientation tuning and its dynamics of cat's primary visual cortex neurons located in pinwheel centers and orientation columns were studied during development of response in time. Functional domains in the Cortex were revealed by intrinsic signal optical imaging on functional and vascular maps of cortex. Both maps were used for precise placement of electrodes during recordings. Orientation tuning dynamic of each neuron was analyzed by temporary slices technique. Several types of neurons were observed in different functional domains: neurons with stable preferred orientation (14.5%); cells with preferred orientation dynamically shifted by 102 +/- 5 degrees (40.8%) and neurons with mixed dynamics type (44.7%). In pinwheel centers neurons with stable preferred orientation are differed from neurons with unstable preferred orientation in their features. In particular, they are showing greater response strength for preferred orientation. In orientation columns neurons with stable tuning are demonstrating larger latent periods for all orientations including preferred and non-preferred in comparison to cells with unstable response dynamics. We observed better detection quality of stable orientation neurons than in unstable preferred orientation neurons. Further coding features of neurons belonging to one functional type but located in different domains were compared. In general, located in pinwheel centers stable neurons have shorter latencies and stronger responses than for the stable cells located in orientation columns. Detection features were similar for this functional type of neurons. Unstable neurons located in different functional domains don't differ from each other in orientation tuning characteristics. Functional significance of neurons with stable and unstable preferred orientation located in pinwheel center and orientation columns in visual information processing are discussed.

[Neurodegenerative changes in rat hippocampal areas during the pentylenetetrazole kindling development].

The development of pentylenetetrazole-induced kindling is accompanied by neurodegeneration and neuronal loss in different areas of the hippocampus. However, the data on neurodegeneration development in the dentate gyrus remain controversial. In our study that was performed on 20 Wistar male rats, it was found that the process of neuronal loss was expressed unequally along the dentate gyrus. By the end of pentylenetetrazole kindling development, degenerating cells were present in the superior and inferior blades of the dentate gyrus, whereas the neuronal density in these areas was not reduced. On the other hand, in the angle of the dentate gyrus neuronal loss was already detected at the very early stages of kindling development. These findings allow to suggest a functional heterogeneity of a population of granule cells in relation to their susceptibility to seizure-induced injury.

[Alpha- and beta-oscillations of dynamics of the area and weight of receptive fields of cat striate neurons under conditions of classical and combined mapping].

In 22 acute experiments in anesthetized and immobilized adult cats the dynamics of 83 receptive fields (RF) of 47 striate neurons was studied by temporal slices method. Classical mapping revealed wave-like changes in the area and weight of neuronal RFs. Special mathematical analysis showed that such changes represented a sum of a slow non-oscillatory and comparatively fast components. The slow component was a biphasic up and down RF dynamics. In most cases, the oscillation frequencies were within the alpha- and beta- EEG frequency ranges. When the RF center was activated additionally during combined mapping, the oscillations frequencies remained unchanged, but the duration and amplitude of non-oscillatory component substantially decreased. Mechanisms underlying the RF dynamics and its functional significance are discussed.

An early decrease in cell proliferation after pentylenetetrazole-induced seizures.

There are increasing data on the influence of seizures on neurogenesis in the adult brain. However, data on cell proliferation and differentiation during the early stages of kindling are scarce. We have used pentylenetetrazole (PTZ)-induced kindling to investigate the temporal profile of cytogenesis in the germinative zones of adult rat brain. For comparison, we also used a single PTZ-induced generalized tonic-clonic seizure. During kindling development, the density of 5-bromo-2'-deoxyuridine-positive cells demonstrated similar changes in all germinative zones: a dramatic decrease after the first subthreshold PTZ injection, and a gradual increase to the control level following repeated PTZ administration. On the contrary, a single PTZ-induced generalized tonic-clonic seizure was followed by an increase in the number of proliferating cells in both the dentate gyrus and the subventricular zone. These results may indicate the existence of global mechanisms affecting cellular proliferation in adult brain during seizures. Different temporal profiles of neuronal damage and proliferation changes suggest that neurodegeneration is unlikely to be a global proliferation-regulating factor. The data may contribute to better understanding of the initial phase of kindling development and epileptogenesis.

[Evaluation of antiepileptic effects of cortexin in a model of convulsions].

We studied antiepileptic effects of cortexin administered in doses 0,015, 0,15 and 1,0 mg/kg intraperitoneally in solution or intranasally in the complex with nanoparticles in a model of acute and chronic convulsions in rats induced by pentylenetetrazole. In the model of epileptic status, the long-term preliminary administration of cortexin had no effect on convulsions while in the model of chronic convulsions (temporal epilepsy), cortexin had a marked dose-dependent antiepileptic effect. The influence of cortexin on neuroplasticity and its clinical potential are discussed.

[Receptive fields of cat striate neurons: dynamics of weight and topographical characteristics of the excitatory zone of the neuron receptive fields in the visual cortex].

In 22 acute experiments on anesthetized and immobilized adult cats, the dynamics of 83 on- and/or off-receptive fields (RF) was studied in 47 striate neurons with the method of temporal slices by mapping with single-flash stimuli. The latency of the RF appearance was 88 +/- 5 ms, and its duration was 192 +/- 12 ms. Dynamical wavy changes in the RF sizes and weights repeating up to three times were revealed. The mean duration of one wave was 95 +/- 4 ms. In 99% of cases, the RF discharge center defined for one temporal slice also displaced in an undulatory way relatively the RF's center defined for whole analysis period. The mean duration of this undulatory cycle was 67.3 +/- 3 ms. In 72.5% of cases, the displacement of the discharge center followed different trajectories inscribed in an ellipse. The functional significance of such changes in the RFs of striate neurons is discussed with regard to the dynamics of their detector features and underlying mechanisms.

[Receptive fields of detectors of bar and cross-like figures in classical and combined mapping].

Excitatory and inhibitory zones of receptive fields (RF) of detectors of cross-like figures and bars were studied in a primary visual cortex of cats under conditions of classical and combined mapping. Factor and correlation analysis demonstrated that detectors of bars and crosses differ in those interrelations, which are formed between the basic characteristics of neurons and their RF in each of group. The factor analysis of these connections showed that variables with the greatest weights, combined into the same factor, are different for different detectors. In detectors of bars, positive correlation was found between the background activity and weight characteristics of the RF excitatory and inhibitory zones. In detectors of crosses, inhibition indices showed a positive correlation with sensitivity to this figure, characteristics of excitatory RF zone and the background activity. Enlargement of the additional excitatory zones of cross detectors in combined mapping was significantly greater than in bar detectors. Formation of the RF and orientation sensitivity of bar and cross detectors is discussed, with account made of the finding that the sensitivity to bars is mostly formed by direct excitatory connections, whereas sensitivity to cross-like stimuli is formed by direct and recurrent intracortical inhibitory connections between the nearby neurons.

Changes in cell proliferation in the subventricular zone of the brain in adult rats given beta-amyloid peptide (25-35).

The effects of intracerebroventricular administration of fragment (25-35) of beta-amyloid peptide [Abeta(25-35)] on cell proliferation in the subventricular zone of the dentate gyrus of the hippocampus in adult rats were analyzed. Animals received doses of 15 nmol of pre-aggregated Abeta(25-35) or the Abeta(35-25) control peptide, or solvent (sterile water) into the lateral ventricles. On post-injection days 1-5, rats received intraperitoneal injections of the thymidine analog 5-bromo-2'-deoxyuridine (BrdU). BrdU incorporated into DNA was detected immunohistochemically on frontal brain sections six and 12 days after peptide administration. At six days, the numbers of BrdU-containing cells in the subventricular zone showed no differences between the study groups. At 12 days, the total number of BrdU-positive cells decreased significantly in all study groups. At the same time, the number of labeled cells in rats given Abeta(25-35) was significantly greater in this brain zone than in animals given water or the control peptide. Thus, Abeta(25-35) significantly increased cell proliferation in the subventricular zone after intracerebroventricular administration.

[Striate receptive fields mapped with single and bipartite stimuli].

In 22 acute experiments with anesthetized and immobilized adult cats, 364 maps of receptive fields (RF) of 47 striate neurons were obtained by means of single local stimuli flashed at different parts of the visual field, or with additional asynchronous activation of the RF excitatory center with oscillating bar of the optimal orientation. Under bipartite stimulation, considerable and significant decrease in the square and weight of the central excitatory RF zone was revealed in more then 75% of the studied cells. Additional excitatory zones appeared in 54% of cases, or the square and weight of the excitatory zones substantially increased, and inhibitory zones developed in 90% of cases. These effects were correlated with the degree of increase in the background firing during transition from the mode of mapping with single stimulation to that with bipartite stimulation. The mechanism and possible functional role of cooperative excitatory and inhibitory intracortical interactions in organization of receptive fields and detection of features of a visual image are discussed.

Neurophysiological and simulation studies of striate cortex receptive field maps: the role of intracortical interneuronal interactions.

Acute experiments on 27 adult anesthetized and immobilized cats investigated 101 on and off receptive fields in 67 neurons in visual cortex field 17 by mapping using single local stimuli presented sequentially at different parts of the visual field, as well as in combination with additional stimulation of the center of the receptive field. Both classical and combined mapping identified receptive fields with single receptive zones (63.4% and 29.3% respectively), along with fields consisting of several (2-5) excitatory and/or inhibitory zones (36.6% and 70.7%). We provide the first report of receptive fields with horseshoe, cross, and T shapes. Simulations of horizontal interneuronal interactions in the visual cortex responsible for the multiplicity of excitatory and inhibitory zones of receptive fields were performed. A role for cooperative interactions of neurons in this effect was demonstrated. The possible functional role of receptive fields of different types in extracting the features of visual images is discussed.

Characteristics of the responses of visual cortex neurons with sensitivity to bars or cross-shaped figures in cats.

The magnitudes and latent periods of spike responses were recorded from 280 individual neurons tuned to the orientation of light bars or cross-shaped figures in the primary visual cortex (field 17) of the cat. In control experimental conditions, half of 195 cells preferred the bar (first group), the remainder preferring crosses (second group); the responses of neurons of the first group to bars and crosses were of similar magnitude, while in the second group, responses to crosses were significantly larger than responses to bars. The latent periods of responses to optimal bars in the first group of neurons were shorter than those in the second group, and became longer on exposure to crosses, while latent periods in the second group were shorter on exposure to crosses. In conditions of local bicuculline blockade of intracortical inhibition, about a quarter of 85 neurons were sensitive only to the bar, regardless of the presence or absence of inhibition. The remaining neurons were sensitive to crosses in at least one of the states and continued to have responses which were smaller in terms of absolute magnitude than the responses of group 1 neurons. The significance of these data for understanding the mechanisms of tuning of striate neurons to signal features and the temporal sequence of their operation is discussed.

[Neurophysiological and simulation study of the striate receptive fields maps: the role of intracortical interaction].

In 27 acute experiments with anesthetized and immobilized adult cats 101 maps of receptive field (RF) in 67 striate neurons were studied by means of mapping with single flashed stimuli presented in different parts of the visual field and under conditions of additional activation of the RF excitatory center by the local oscillating or flashing grid. Under conditions of both classical and combined modes of mapping, the RFs of the classical shape with a single excitatory zone (63.4 and 29.3% of cases, respectively) and RFs with multiple (2-5) excitatory and/or inhibitory zones (36.6 and 70.7%, respectively) were found. We were the first to describe, also, some RFs of horseshoe-like, cross-like and T-like shapes. Simulation of non-classical RFs revealed possible contributions of cooperative excitatory and inhibitory intracortical interactions to the effects under study. The functional role of RFs of different types in the feature detection is discussed.

[Nitric oxide synthase activity and nitrates/nitrites level in brain regions during the spontaneous morphine withdrawal in rats].

Nitric oxide synthase (NOS) activity and nitrate/nitrites (NO(x)-) concentrations were measured in brain regions of rats during the spontaneous morphine withdrawal. Male Wistar rats were injected intraperotoneally with morphine hydrochloride at increasing doses (10-100 mg/kg) during 6 days twice a day. Thirty six hours after the last injection the severity of the spontaneous morphine withdrawal syndrome was determined by specific autonomic and locomotor indices. Both NOS activity and NO(x)- levels increased in the midbrain and the hippocampus, decreased in the striatum and the hypothalamus, and did not change in the cerebral cortex and the brain stem. NO(x)- concentrations in the cerebellum did not change, while NOS activity decreased. Both NOS activities and NO(x)- concentrations in the cerebral cortex, striatum, midbrain, and cerebellum correlated with withdrawal syndrome severity on the whole, and with the specific signs of abstinence.

[Responses of neurons in cat visual cortex with sensitivity to bar and cross-like figure].

In the cat primary visual cortex (area 17) the response magnitude and latency were studied in 280 neurons sensitive to bar or cross-like-figure. Under natural conditions half of the studied 195 cells preferred bar (first group) or cross (second group). In the first group responses to both figures were near equal, while in the second one cross evoked much stronger response. Response latencies with the optimal bar in the first group were shorter than in the second group and longer to a cross than to a bar while in the second group they were considerably shorter to a cross than to bar. Under local blockage of GABA-ergic inhibition by microiontophoretic application of bicuculline about one-fourth of 85 neurons generated greater responses and were bar-sensitive irrespective to presence or absence of inhibition. Other neurons were cross-sensitive at least in one of the conditions (with and/or without of inhibition). They responses grew under bicuculline action relatively more than in the first group. Significance of the data obtained for tuning to image features and temporal succession of their detection is discussed.

[Effects of intracerebroventricular administration of a caspase-3 inhibitor Z-DEVD-FMK on behavior of rats].

Rats received intracerebroventricular injections of z-DEVD-FMK (caspase-3 inhibitor) or z-FA-FMK (control peptide) in a dose of 3 nmol. Administration of z-DEVD-FMK significantly decreased the number of avoidance reactions in some blocks of trials in active avoidance (shuttle box) learning. However, only slight effect of the caspase inhibitor across the session was found. Z-DEVD-FMK impaired development of some essential components of the two-way active avoidance performance, such as escape reaction, conditioned fear reaction, and inter-trial crossings. Z-DEVD-FMK did not impair working memory in the spontaneous alternation behavior paradigm. Z-DEVD-FMK affected neither emotionality nor locomotor activity in the open-field test. It also did not influence behavior in the light-dark chamber. Measurement of caspase-3 activity in rat brain regions involved in active avoidance learning revealed z-DEVD-FMK-related inhibition of the enzyme activity most pronounced (about 30%) in the fronto-parietal cortex; a similar effect was close to significant in the hippocampus. The results suggest the involvement of brain caspase-3 in selected forms of learning.