Participation of frontal lobe neurons in the organization of successive inhibition and disinhibition of conditioned reflexes in the dog

Neuronal activity in g. proreus was studied during classical secretory conditioning and its differentiation in dogs. Three types of changes in neuronal reaction pattern were identified during differential conditioned stimulus compared to the pattern observed during positive conditioned stimulus. It has been shown that signal significance of the conditioned stimulus may be coded by specific distribution of interspike intervals in response to different conditioned stimuli. In situation of successive inhibition or disinhibition of the differentiation some neurones displayed simultaneous appearance of interspike intervals specific for responses to current and previously presented conditioned stimuli.     

Spectral-correlative characteristics of the EEG reactions preceding conditioned reflex motor reactions in dogs

The work is a logical continuation of previous studies (analysis of the background electrical activity in the band 1-100 Hz in interstimulus intervals in the process of lever pressing alimentary conditioning in dogs) and it is dedicated to correlation-spectral analysis of prestimulus periods and EEG-reactions to conditioned stimuli, previous to conditioned lever pressing. Visually the EEG reactions present discharges of high-frequency (40-100 Hz) synchronized activity preceding for 40-300 ms the beginning of the changes in EMG of the "working" limb. It is shown that EEG reactions are characterized (in comparison with the background activity) by a higher energetic level and a greater expression of the high coherence (I greater than 0.75) and also by greater phase shifts, in counterbalance to the domination of little phase shifts in the background activity. It is assumed that the patterns of EEG reactions may participate in trigger mechanisms either eliciting conditioned motor reactions (to positive conditioned stimuli) or preventing them (to inhibitory conditioned stimuli).     

The dependence of the neuronal reactions of the sensorimotor cortex to a simultaneous complex stimulus on the level of the differentiation of its components]

On alert animals the change was studied of the neuronal activity of the sensorimotor cortical area of cats brain in dependence on the level of differentiation of the components of simultaneous heteromodal complex stimulus. According to the character of this dependence and a number of other parameters two groups of neurones were singled out in the sensorimotor cortex. It was shown that parameters of reactions of all recorded neurones of the sensorimotor cortex to the positive conditioned signal were the first established after consolidation of the animal conditioned motor activity. In the course of elaboration following parameters changed: expressiveness, intensity, duration and value of latency. Reactions of neurones of both groups to the inhibitory stimuli were stabilized only after consolidation of the habit of differentiation. Responses of the first group neurones changed only by the pattern of discharge, while the responses of the second group neurones could change by expressiveness of response, its sign, duration and value of latency. Oscillations of the differentiation level after finishing of the elaboration of inhibitory conditioned reactions affected only the responses of the second group neurones to complex components.     

Neuron-Like Differentiation of PC12 Cells Treated With Media Conditioned by Either Sciatic Nerves,Optic Nerves,or Schwann Cells

In previous works we reported the finding of neurotrophic activity in a serum-free Dulbeccos modified Eagles medium conditioned by rat sciatic nerves, previously maintained in culture for 11 days. This medium produces rapid neuron-like differentiation of cultured PC12 cells, as revealed by an increase in the size of the cell body and by the extension of short and/or long neurites by most of the cells. Neuregulin present in the conditioned medium was demonstrated to play a key role in the observed differentiation.In the present work, taking into consideration those latter results, the neurotrophic activity of conditioned media prepared with sciatic and optic nerves cultured during days 1–4 and 9–12 were studied.Evaluation of the trophic activities of those media revealed an opposite timing in the activities of sciatic and optic nerves conditioned media. The activity of the sciatic nerve was not observed in the 1–4-day period, increasing then up to the 9–12-day period. On the contrary, the optic nerve conditioned medium was active in the 1–4-day period, decreasing down to the 9–12-day period.These results led us to explore the contribution of the different cellular constituents of those nerves to their neurotrophic properties. As a first step in that direction we also investigated the neurotrophic activity of media conditioned during 12 days by cultured Schwann cells isolated from rat sciatic nerves. The Schwann cell conditioned media did produce a rapid differentiation of the PC12 cells similar to that caused by the sciatic nerve conditioned medium, though of a lower magnitude.Variations in the trophic activities of the conditioned media used in the present work is discussed taking into consideration the production of trophic and inhibitory factors by the peripheral and central glial cells. The role played by the optic nerve glia and myelin is being investigated at present.     

Effect of activation and blockade of the GABA-ergic system of the substantia nigra in the midbrain on the realization of conditioned food reflexes in dogs

Bilateral injection of 45 mcg of GABA into substantia nigra pars compacta produced in dogs a manifested improvement of parameters of the conditioned differentiation inhibition but failed to influence the positive Pavlovian alimentary conditioned reflex. Injection of GABA synaptic antagonist--picrotoxin impaired conditioned alimentary behaviour. Numerous injections of the GABAergic pharmacological agents resulted in motor disturbance--rotatory movements--and skin trophic deviations. The data obtained and literature references give ground for discussion of the role of striato-nigral and internal GABAergic substantia nigra systems in the positive modulation of adaptive alimentary behaviour and conditioned stimuli differentiation.     

Reversal of signal significance of stimuli in the avoidance reflex]

The course of reversal learning in go -- no go avoidance reflex differentiation in both cats and dogs was analysed in terms of interrelations between drive and instrumental reflex activity. When signalling properties of the conditioned stimuli were reversed, marked changes in the drive state of experimental animals occurred. The increase of the fear drive influenced both the transfer of the instrumental response to the new positive conditioned stimulus and the extinction of the response to the previously positive stimulus. The quality of the conditioned stimuli and their reflexogenic strength exerted clear effects on the fear drive and on the course of reversal learning. Prefrontal lesions affected the drive state and the conditioned reflex activity of the animals during the reversal learning.     

Effect of neurotizing exposures on conditioned reflex activity in the late postresuscitation period in rats

In white rats resuscitated after 10-minutes stop of the blood circulation, conditioned switching over was elaborated on the basis of alimentary and defensive instrumental reflexes and the effect was studied of neurotizing influences. Single application of electrocutaneous pain stimulus in alimentary situation ("collision") led to more expressed disturbances of conditioned activity in resuscitated rats than in intact ones. Differentiation of a signal different from the positive conditioned stimulus only by the height of the tone, was practically not elaborated in resuscitated rats, and the disturbance of the conditioned alimentary reflex to the reinforced signal appearing during differentiation elaboration, was manifest in them to a greater degree. The obtained data testify that even a short-time brain ischemia, accompanied by a fast and full restoration of the neurological status, leads to functional defects of the CNS appearing at subsequent neurotic influences.     

Typological characteristics of conditioned reflex learning in monkeys

Three types of conditioned learning dynamics were singled out in males of the rhesus monkeys by the speed of formation of a positive motor instrumental reflex and differentiation. Type I is characterized by the fastest mastering of separate subprograms and of the whole test program of conditioned behaviour, by optimal functional characteristics in the process of learning and by a high adaptivity in new conditions. The most prolonged formation of the conditioned reflexes occurs in monkeys of the III-d type; 50% of them do not master the whole learning program; monkeys with such type of learning dynamics have no adaptation of previously acquired conditioned stereotype to new environmental conditions. Monkeys of the II-nd type of learning dynamics possess intermediate characteristics. A notion is substantiated of interconnection of higher nervous activity properties with typological characteristics of conditioned learning dynamics in monkeys.     

Effect of microinjections of a selective blocker of M1-muscarinic receptors pirenzepine into the neostriatum on the rat motor activity

In simulated discrimination conditioned reflex of active avoidance (CRAA) in T-maze, the effect of bilateral microinjections of the muscarinic receptor M1 selective blocker pirenzepine on the CRAA formation and behaviour in the "open filed" test, was studied in rats. A sharp worsening of the CRAA learning and a significant increase in the motor activity were shown to occur in rats following the microinjections as compared with control rats. The change in the motor responses seems to account for the worsening of the CRAA learning. Another reason of the phenomenon could involve a disorder in perception of conditioned signals and their poor differentiation. The data obtained and the literature data suggest a complex character of changes induced by the blockade of the M1 muscarinic receptors of the neostriatum.     

Differentiation of a cell line of mouse myeloid leukemia : I. Simultaneous induction of lysosomal enzyme activities and phagocytosis

Induction of phagocytic activity in the Ml cell line of mouse myeloid leukemia, on being exposed to a conditioned medium from cultured embryo cells, was accompanied by an increment in the activities of both lysosomal acid phosphatase and acid protease. The activity of these lysosomal enzymes, as well as that of phagocytosis, was not induced when Ml cells were incubated either with the conditioned medium subjected to heat treatment or in the presence of 5-bromodeoxyuridine (BUdR). The levels of these induced enzyme activities in Ml cells were comparable to those in normal mouse peritoneal macrophages. The lysosomal enzyme activity in Mm-1 cells, which were spontaneously differentiated from Ml cells and exhibiting a higher phagocytic activity, were reminiscent of those in peritoneal macrophages. Based on these observations, it was concluded that both phagocytosis and lysosomal enzyme activity occur simultaneously during the course of differentiation. This differentiation, morphological or functional, in Ml cells in the presence of the conditioned medium was further supported by biochemical evidence.     

Experimental neuroses in dogs caused by an increase in the intensity of (light) stimuli conditions

The influence of enhanced intensity of general illumination on conditioned reactions of a different degree of complexity was studied on dogs by the classical method of secretory conditioned reflexes. In most of the dogs, increase of general illumination from 30 to 300 luxes leads to more intensive secretory conditioned and unconditioned reactions, a smaller number of errors in differentiation, and stabilization of parameters of conditioned activity, which testifies to the improved general functional state of the brain. In two animals in the same conditions of the experiment, an experimental neurosis set it, not connected with the complexity of the elaborated activity. It is assumed that the neurotizing agent was the superfluous situational (photic) stimulation which presented excessive requirements to the mechanisms regulating the general functional state of the brain.     

Slow potentials of the prefrontal cortex in dogs and the classical secretory conditioned reflex

Slow potentials (CNV and component P300) were recorded in the medial part of the prefrontal cortex of dogs trained to classical secretory conditioned reflex and its differentiation. CNV increased when the conditioned stimulus was preceded by a signal of different meaning, as compared with CNV to the same conditioned signal following the stimulus of the same meaning; the greatest values of CNV and P300 were observed in response to differential stimulus preceded by a positive signal.     

A factor produced by feeder cells which inhibits embryonal carcinoma cell differentiation. Characterization and partial purification

Medium conditioned by STO mouse fibroblast cells inhibited both the spontaneous differentiation of NG2 embryonal carcinoma cells and the differentiation of F9 embryonal carcinoma cells induced by retinoic acid. This effect was due to a differentiation retarding factor (DRF). Reduction in DRF activity in conditioned medium by boiling and by pronase treatment suggested the involvement of a polypeptide, which had an apparent molecular weight of 57000 on gel filtration. A 28-fold purification of DRF was achieved. DRF delayed but did not prevent the extensive differentiation observed after prolonged culture of NG2 colonies. Conditioned medium could be successfully used to replace feeder cells in NG2 stock cultures. Media conditioned by a variety of other cell types also contained differentiation retarding activity.     

Features of the formation and dynamics of short-term image memory in the rat

Direct and indirect methods of delayed reactions showed that short-term image memory of rats elaborated on the basis of conditioned signals in poorly developed and its duration is not more than 1 s. While testing this memory, changes in the behaviour were seen, pointing to the development of "hard state" in the higher nervous activity of animals. Short-term image memory based on complex perception was shown to be well developed in rats, and by many characteristics it approximated to the memory of highly organized animals. The ability to reproduce the short-term image memory on the basis of conditioned signals is supposed to be a complex form of psychonervous activity of animals, and in rodents it appears to be at an initial stage of development.     

Purification of a factor inhibiting differentiation from conditioned medium of nondifferentiating mouse myeloid leukemia cells

Mouse myeloid leukemic M1 cells are induced to differentiate by various differentiation inducers. Activity for inhibition of induction of differentiation of M1 cells (I-factor activity) was detected in conditioned medium of variant M1 cell clones that were resistant to differentiation inducers, and this I-factor activity was shown to be closely associated with resistance of the cells to differentiation inducers. In this work, the I-factor was purified to apparent homogeneity from conditioned medium of resistant M1 cells. The purification procedure consisted of ammonium sulfate precipitation, CM-Sepharose CL-6B, Sephadex G-200, reverse-phase high performance liquid chromatography on a C18 hydrophobic support, and high-performance liquid chromatography on a gel filtration column. The factor was analyzed by radioiodination, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and autoradiography. The purified factor gave a single band of protein with a molecular weight of 68,000 on sodium dodecyl sulfate-polyacrylamide gel electrophoresis which coincided with its biological activity. The concentration of I-factor required for 50% inhibition of dexamethasone-induced differentiation of M1 cells was 24 pM. At its effective concentration it had no effect on cell proliferation, and even at 1.2 nM it did not inhibit colony formation of normal bone marrow cells, suggesting that it was distinct from the inhibitor of normal precursors of macrophages and/or granulocytes.     

Neuronal activity of the cerebral cortex in cats with conditioned inhibition

Unit activity of the somatosensory cortex was studied in chronic experiments on cats during alimentary instrumental conditioned reflex performance and elaboration of conditioned inhibition (CI). First presentations of the CI signal at the beginning of CI formation caused, due to orienting reaction to a new stimulus, a levelling of the response of the neurones to the positive conditioned signal included into the CI complex. At the stage of consolidated CI, this depression proceeded gradually during the development of conditioned inhibition caused by consecutive presentations of the nonreinforced CI combination. Two groups of neurones were involved in the process of the CI performance: one of them being the same which was activated also in response to positive stimulation during performance of the conditioned response, and the second one being areactive to the positive conditioned stimulus but reducing the frequency of the background activity.     

A possible mechanism of participation of dopaminergic cells and striatal cholinergic interneurons in the conditioned selection of motor activity

A possible mechanism of participation of cholinergic striatal interneurons and dopaminergic cells in conditioned selection of a certain types of motor activity is proposed. This selection is triggered by simultaneous increase in the activity of dopaminergic cells and a pause in the activity of cholinergic interneurons in response to a conditioned stimulus. This pause is promoted by activation of striatal inhibitory interneurons and action of dopamine at D2 receptors on cholinergic cells. Opposite changes in dopamine and acetylcholine concentration synergistically modulate the efficacy of corticostriatal inputs, modulation rules for the "strong" and "weak" corticostriatal inputs are opposite. Subsequent reorganization of neuronal firing in the loop cortex--basal ganglia--thalamus--cortex results in amplification of activity of the group of cortical neurons that strongly activate striatal cells, and simultaneous suppression of activity of another group of cortical neurons that weakly activate striatal cells. These changes can underlie a conditioned selection of motor activity performed with involvement of the motor cortex. As follows from the proposed model, if the time delay between conditioned and unconditioned stimuli does not exceed the latency of responses of dopaminergic and cholinergic cells (about 100 ms), conditioned selection of motor activity and learning is problematic.     

Patterns in the transition from normal to neurosis in dogs with different typological traits in food and defensive situations

At comparison of the formation of neurosis in dogs with different typological characteristics in conditions with alimentary and pain reinforcement, a resemblance was revealed of transition from norm to the initial stage of pathology. In strong animals a development was seen of compensatory tachycardia at circular changes of conditioned activity. Weak type dogs are characterized by a lowering or full disappearance of reflexes in combination with bradycardia. The method of differentiation, allowing in situations with alimentary reinforcement to classify the tested animals according to the type of higher nervous activity and differentiate the states of "norm" and "initial stage of neurosis", proved to be effective for the analogous aims also in studies with defensive reinforcement.     

Modeling neurosis in the dog and a study of the effects of antidepressants

Introduction in tests with positive (alimentary) reinforcement, of elements of uncertainty and of "collision" of motivations, and carrying out of a series of experiments on dogs under such conditions during 2-4 weeks, led to the development of heavy and stable neurotic states. They were characterized by disturbances of emotional-motivational sphere, suppression of conditioned activity, somatovegetative dysfunctions. Preparations of atypical antidepressants (emovit and new benzamine derivate) reduced neurotic manifestations. The rate of reduction depended on the spectrum of neuro-psychotropic activity and properties of pathological states in dogs.     

Changes in conditioned reflex activity during adaptation of the rat to pressure-chamber and high-altitude hypoxia

A group of rats with a stereotype of conditioned reflexes was preliminarily trained to hypoxia effects during 30 days (at the "altitude" of 6000 m, time of exhibition--from 10 to 60 min, for 18 days--only 60 min). Adaptive changes in the process of training consisted in a weakening of differentiation inhibition, partial amnesia of the conditioned reaction of active avoidance and appearance of phasic states (equalization and paradoxical phases) in the cerebral cortex. The following adaptation of hypoxia "trained" rats to new natural conditions of Alpine altitude (3200 m) proceeded favourably, without disturbance of differentiation inhibition and without phasic states. Rats without preliminary training to altitude chamber hypoxia, in mountains (3200 m) were subjected to moderate tension resulting in protective inhibition, partial amnesia and transient disturbance of differentiation inhibition.