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The prevalence of autism spectrum disorder (ASD) has rapidly increased in the past decades, and several studies report about the escalating use of antibiotics and the consequent disruption of the gastrointestinal microbiome leading to the development of neurobehavioral symptoms resembling to those of ASD. The primary purpose of this study was to investigate whether depletion of the gastrointestinal microbiome via antibiotics treatment could induce ASD-like behavioral symptoms in adulthood. To reliably evaluate that, validated valproic acid (VPA) ASD animal model was introduced. At last, we intended to demonstrate the assessed potential benefits of a probiotic mixture (PM) developed by our research team. Male Wistar rats were used to create antibiotics treated; antibiotics and PM treated; PM treated, VPA treated; VPA and PM treated; and control groups. In all investigations we focused on social behavioral disturbances. Antibiotics-induced microbiome alterations during adulthood triggered severe deficits in social behavior similar to those observed in the VPA model. Furthermore, it is highlighted that our PM proved to attenuate both the antibiotics- and the VPA-generated antisocial behavioral symptoms. The present findings underline potential capacity of our PM to improve social behavioral alterations thus, indicate its promising therapeutic power to attenuate the social-affective disturbances of ASD.

In Hungary, cancer causes a quarter of all deaths. Our goal is to develop a method of early diagnosis. Our research team is developing a new non-invasive bioimpedance spectrometry-based procedure that enables early detection of cancers. The measurement procedure as well as the instruments used for the measurement have been specially developed to measure tumor cells. The system, which is entirely self-developed, is an innovative solution for eliminating contact impedance due to the agility of the ions during the two electrode measurements known to this time. In our article, we present a cell culture plate that generates data that is free from the distractions of the aforementioned contact impedance. During the physical validation, we have demonstrated that the methodology we have developed can be used in a wide frequency range, so it is therefore fully suitable for further cellular testing.

In recent times, our knowledge on the gastrointestinal microbiota has considerably improved. Dysfunctions of the gut microbiome have been shown to induce modifications of peripheral and central regulatory processes, ultimately leading to behavioral changes that cause functional deficits in brain functions. Therefore, the first aim of the present study was to generate massive dysbiosis of the gut microbiome and to investigate these effects on behavioral responses in adulthood. To determine the impact of the alterations on the behavior we used adult male Wistar rats. Animals have been divided into two groups – the first one was an antibiotics treated-and the other one was a control group. As antibiotic treatment, rats were given broad spectrum antibiotic mixture for 4 weeks. Substances were dissolved in the animals’ drinking water. Following the intestinal microbiome depletion. Open filed test (OFT) was conducted. Throughout the whole experiment, body weight, food and water consumptions were daily monitored, and faecal samples were collected for later determination of the short chain fatty acid (SCFA) levels. The findings demonstrated significant group-differences such as abnormal behavioral and metabolic phenomena identified among the antibiotic treated animals unraveled in the behavioral test as well as in the short chain fatty acid analyses. Our results reinforce that alterations of the microbiota play important modulating role in the central regulatory processes. However, it is uncertain whether the antibiotic-induced dysbiosis directly triggers the altered behavioral phenomena, or these might have caused the change of body composition which itself could define the behavior. Exploring this question will be our next step to determine how the alterations of microbiome can affect behavioral patterns. We aim to introduce a non-invasive body composition measurement technique to explore this question. This method based on the electrical impedance spectrum analysis. By means of this new four-electrode ultra-low frequency bioimpedance measurement technology, we can carry out high-precision full-body measurements or we could correlate bioimpedance measurement data to various parts of the body if it required. By using the bioimpedance technology, we did not observe significant body composition differences between the two groups. These results confirm our hypothesis that antibiotic-induced dysbiosis directly influences the control of behavioral processes.

The oxygen plays an important role in the development and function of the human brain. In the central nervous system, a different neuron types can be very sensitive to varying concentrations of oxygen. Despite several cell biology researches, the time course of the effects of non-physiological oxygen concentrations, but especially high oxygen concentrations on the membrane of neurons, and the dynamics of damage are unknown. In the present study, with this in view, electrical impedance spectroscopy (EIS) measurements are carried out using a graphene-based self-developed four-electrode measurement and data collection process. Our goal was to produce a plate layout for our further experiments that could be used to examine cell cultures under hypoxic and hyperoxic conditions. The designs differ in the distance between the electrodes, we used three types of distances: 1 mm, 3 mm 5 mm, using one distance per plate. On the plate with 5 mm electrode spacing we found the ideal. From a measurement point of view, the plate with 5 mm electrode spacing was the most suitable. As we increased the distance, the size of the electrode surfaces decreased. It can be assumed that reducing the size of the electrodes did not negatively affect the measurement results. Our results can greatly contribute to better understand the process of cell membrane damage.

Nowadays, the gold standard for the diagnosis of liver disease is histological examination. However, due to the dangers and complications of liver biopsy, there is a worldwide demand for the development of non-invasive procedures that can accurately determine the degree of hepatic steatosis, furthermore is cost-effective, user-friendly, and provides repeatable measurement. Based on the metrological experience of our research team, we started developing an electrical impedance-based diagnostic method a few years ago that meets these requirements and is suitable for early detection of nonalcoholic fatty liver disease (NAFLD) with the help of a worldnew measurement and data collection solution. In addition to the preliminar results of the long-term clinical validation of the diagnostic method developed to detect NAFLD, the authors present early partial results. Even with the small amount of data, very impressive results have been obtained.

Compacted neurons are morphologically changed axons in an undamaged environment, which are able to recover spontaneously. They may play an important role in neurological disorders, such as Parkinson’s disease or Alzheimer’s disease. The aim of this study was to find the best method for the investigation of this process. Therefore we designed and selfmade bio-impedance spectrum (BIS) measurement plates with three different electrode distances (1mm, 3mm, 5mm). Due to measurements using self-developed software, we got the result that plates with 3mm and 5mm distance setups are suitable for our future research studies. The validation measurement was carried out with series of solutions made of ion exchanged water and physiological saline solution (concentrations: 0%, 25%, 50%, 75%, and 100%) Due to successful plate development, it became possible to continuously measure the process of compaction and recovery of the dark neurons, and find out the details of this phenomenon.

Non-alcoholic fatty liver disease (NAFLD) is a leading cause of chronic liver disease worldwide, it is the primary cause of liver-related morbidity and mortality. The prevalence of the disease is estimated as high as 25% of the general population up to 90% of obese and diabetic patients. Most patients are asymptomatic or does not experience specific complaints making them difficult to diagnose and treat. However, due to the limitations of available diagnostic procedures and the current gold standard for diagnosis, the liver biopsy, there is a worldwide demand to develop a non-invasive diagnostic tool and procedure that can detect the disease with high accuracy at an early reversible stage, furthermore is cost-effective, user-friendly, and provides repeatable measurement. Based on the metrological experience of our research team, we started developing an electrical impedance-based diagnostic procedure a few years ago that meets these requirements and is suitable for early detection of NAFLD with the help of a world-new measurement and data collection solution. In this article, we present the most important milestones in the development of instruments related to research.

A central goal of systems neuroscience is to simultaneously measure the activities of all achievable neurons in the brain at millisecond resolution in freely moving animals. This paper describes a protocol converter which is part of a measurement acquisition system for multichannel real time recording of brain signals. In practice, in such techniques, a primary consideration of reliability leads to great necessity towards increasing the sampling rate of these signals while simultaneously increasing the resolution of A/D conversion to 24 bits or even to the unprecedented 32 bits per sample. In fact, this was the guiding principle for our team in the present study. By increasing the temporal and amplitude resolution, it is supposed that we get enabled to discover or recognize and identify new signal components which have previously been masked at a “low” temporal and amplitude resolution, and these new signal components, in the future, are likely to contribute to a deeper understanding of the workings of the brain., Electrical engineering; Neuroscience; Electrical system; Electrical system planning; Systems neuroscience; I2S protocol; TCP/IP protocol; Protocol converter; Scalable system

Effects of kainate or 6-hydroxidopamine (6-OHDA) lesions in the ventromedial prefrontal cortex (vmPFC) on taste-related learning and memory processes were examined. Neurotoxins were applied by iontophoretic method to minimize the extent of lesion and the side effects. Acquisition and retention of conditioned taste avoidance (CTA) was tested to different taste stimuli (0.05 M NaCl, 0.01 M saccharin, 0.01 M citrate and 0.00025 M quinine). In the first experiment, palatability index of taste solutions with these concentrations has been determined as strongly palatable (NaCl, saccharin), weakly palatable (citrate) and weakly unpalatable (quinine) taste stimuli. In two other experiments vmPFC lesions were performed before CTA (acquisition) or after CTA (retrieval). Our results showed that both kainate and 6-OHDA microlesions of vmPFC resulted in deficit of CTA acquisition (to NaCl, saccharin and citrate) and retrieval (to NaCl and saccharin). Deficits were specific to palatable tastants, particularly those that are strongly palatable, and did not occur for unpalatable stimulus. The present data provide evidence for the important role of vmPFC neurons and catecholaminergic innervation of the vmPFC in taste related learning and memory processes.

Tridecapeptide Neurotensin (NT) is widely distributed in the central nervous system where it acts as a neurotransmitter and neuromodulator. The central nucleus of amygdala (CeA), part of the limbic system, plays an important role in learning, memory, anxiety and reinforcing mechanisms. Our previous data showed that NT microinjected into the CeA has positive reinforcing properties. We supposed that these effects might be due to modulations of the mesolimbic dopamine system. The aim of our study was to examine in the CeA the possible effects of NT and dopamine interaction on reinforcement by conditioned place preference test. Male Wistar rats were microinjected bilaterally with 100 ng NT or 2 μg D1 dopamine receptor antagonist alone, or D1 dopamine antagonist 15 min before 100 ng NT treatment or vehicle solution into the CeA. Other animals received 4 μg D2 dopamine receptor antagonist Sulpiride alone, or administration of D2 dopamine receptor antagonist 15 min before 100 ng NT treatment or vehicle solution into the CeA. Rats that received 100 ng NT spent significantly more time in the treatment quadrant during the test session. Pre-treatment with the D1 dopamine antagonist, blocked the effects of NT. D2 dopamine receptor antagonist pretreatment could prevent the positive reinforcing effects of NT as well. Antagonists themselves did not influence the place preference. Our results show that the rewarding effect of NT can be due to the modulation of DA system, since its effects could be blocked by either D1 dopamine or D2 dopamine antagonist preteatment.

Multiple functional attributes of glucose-monitoring neurons in the medial orbitofrontal (ventrolateral prefrontal) cortex. NEUROSCI BIOBEHAV REV 73(1) XXX-XXX, 2017.- Special chemosensory cells, the glucose-monitoring (GM) neurons, reportedly involved in the central feeding control, exist in the medial orbitofrontal (ventrolateral prefrontal) cortex (mVLPFC). Electrophysiological, metabolic and behavioral studies reveal complex functional attributes of these cells and raise their homeostatic significance. Single neuron recordings, by means of the multibarreled microelectrophoretic technique, elucidate differential sensitivities of limbic forebrain neurons in the rat and the rhesus monkey to glucose and other chemicals, whereas gustatory stimulations demonstrate their distinct taste responsiveness. Metabolic examinations provide evidence for alteration of blood glucose level in glucose tolerance test and elevation of plasma triglyceride concentration after destruction of the local GM cells by streptozotocin (STZ). In behavioral studies, STZ microinjection into the mVLPFC fails to interfere with the acquisition of saccharin conditioned taste avoidance, does cause, however, taste perception deficit in taste reactivity tests. Multiple functional attributes of GM neurons in the mVLPFC, within the frame of the hierarchically organized central GM neuronal network, appear to play important role in the maintenance of the homeostatic balance.

Substance P (SP) and neurotensin (NT) are neuropeptides isolated in the periphery and in the central nervous system. They are involved in various regulatory processes in the gastrointestinal tract, in the circulatory and respiratory systems, kidney and endocrine system. In addition to the peripheral effects, SP and NT act as neurotransmitters and neuromodulators in the central nervous system, regulating various behavioural actions, such as general and motor activity, pain, food and water intake, anxiety, reward/reinforcement and memory consolidation. In the limbic system SPergic and NTergic pathways, terminals and related receptors have been identified. According to several data of literature and to our recently published results, SP and NT have rewarding/reinforcing effects and facilitate memory consolidation in various limbic regions. In this report evidences are provided about the interaction of these neuropeptides with dopaminergic and acetylcholinergic systems. A hypothesis is presented that rewarding/reinforcing effects of SP and NT develop by modulating the mesencephalic dopaminergic system, while their mnemonic effects are mediated via the mesencephalic dopaminergic and the basal forebrain cholinergic systems.

In order to elucidate whether cytokine mechanisms of the cingulate cortex (cctx) are important in the central regulation of homeostasis, in the present study, feeding-metabolic effects of direct bilateral microinjection of interleukin-1β (IL-1β) into the cctx of the rat have been investigated. Short- (2 h), medium (12 h) and long-term (24 h) food and water intakes and body temperature were measured after the intracerebral administration of this primary cytokine or vehicle solution, with or without paracetamol pretreatment. The effect of IL-1β on the blood glucose level of animals was examined in glucose tolerance test (GTT), and concentrations of relevant plasma metabolites (total cholesterol, HDL, LDH, triglycerides, uric acid) were additionally also determined following the above microinjections. In contrast to causing no major alteration in the food and water intakes, the cytokine treatment evoked significant increase in the body temperature of the rats. Prostaglandin-mediated mechanisms were shown to have important role in the mode of this action of IL-1β, since paracetamol pretreatment partially prevented the development of the above mentioned hyperthermia. In the GTT, no considerable difference was observed between the blood glucose levels of the cytokine treated and control animals. Following IL-1β microinjection, however, significant decrease of HDL and total cholesterol was found. Our present findings indicate that elucidating the IL-1β mediated homeostatic control mechanisms in the cingulate cortex may lead to the better understanding not only the regulatory entities of the healthy organism but also those found in obesity, diabetes mellitus and other worldwide rapidly spreading feeding-metabolic disorders.

Abstract: Introduction: The medial orbitofrontal cortex is involved in the regulation of feeding and metabolism. Little is known, however, about the role of local glucose-monitoring neurons in these processes, and our knowledge is also poor about characteristics of these cells. Aim: The functional significance of these chemosensory neurons was to be elucidated. Method: Electrophysiology, by the multibarreled microelectrophoretic technique, and metabolic investigations, after streptozotocin induced selective destruction of the chemosensory neurons, were employed. Results: Fifteen percent of the neurons responded to glucose, and these chemosensory cells displayed differential neurotransmitter and taste sensitivities. In acute glucose tolerance test, at the 30th and 60th minutes, blood glucose level in the streptozotocin-treated rats was significantly higher than that in the controls. The plasma triglyceride concentrations were also higher in the streptozotocin-treated group. Conclusions: Glucose-monitoring neurons of the medial orbitofrontal cortex integrate internal and external environmental signals, and monitor metabolic processes, thus, are indispensable to maintain the healthy homeostasis. Orv Hetil. 2017; 158(18): 692–700.

Manganese-enhanced magnetic resonance imaging (MEMRI) offers unique advantages such as studying brain activation in freely moving rats, but its usefulness has not been previously evaluated during operant behavior training. Manganese in a form of MnCl2, at a dose of 20mg/kg, was intraperitoneally infused. The administration was repeated and separated by 24h to reach the dose of 40mg/kg or 60mg/kg, respectively. Hepatotoxicity of the MnCl2 was evaluated by determining serum aspartate aminotransferase, alanine aminotransferase, total bilirubin, albumin and protein levels. Neurological examination was also carried out. The animals were tested in visual cue discriminated operant task. Imaging was performed using a 3T clinical MR scanner. T1 values were determined before and after MnCl2 administrations. Manganese-enhanced images of each animal were subtracted from their baseline images to calculate decrease in the T1 value (ΔT1) voxel by voxel. The subtracted T1 maps of trained animals performing visual cue discriminated operant task, and those of naive rats were compared. The dose of 60mg/kg MnCl2 showed hepatotoxic effect, but even these animals did not exhibit neurological symptoms. The dose of 20 and 40mg/kg MnCl2 increased the number of omissions and did not affect the accuracy of performing the visual cue discriminated operant task. Using the accumulated dose of 40mg/kg, voxels with a significant enhanced ΔT1 value were detected in the following brain areas of the visual cue discriminated operant behavior performed animals compared to those in the controls: the visual, somatosensory, motor and premotor cortices, the insula, cingulate, ectorhinal, entorhinal, perirhinal and piriform cortices, hippocampus, amygdala with amygdalohippocampal areas, dorsal striatum, nucleus accumbens core, substantia nigra, and retrorubral field. In conclusion, the MEMRI proved to be a reliable method to accomplish brain activity mapping in correlation with the operant behavior of freely moving rodents.

In our present experiments, the role of D2 dopamine (DA) receptors of the ventral pallidum (VP) was investigated in one trial step-through inhibitory avoidance paradigm. Animals were shocked 3 times in the conditioning trial, with 0.5 mA current for 1 s. Subsequently bilateral microinjection of the D2 DA receptor agonist quinpirole was administered into the VP in three doses (0.1 μg, 1.0 μg or 5.0 μg in 0.4 μl saline). We also applied the D2 DA receptor antagonist sulpiride (0.4 μg in 0.4 μl saline) alone or 15 min prior to the agonist treatment to elucidate whether the agonist effect was specific for the D2 DA receptors. Control animals received saline. In a supplementary experiment, it was also investigated whether application of the same conditioning method leads to the formation of short-term memory in the experimental animals. In the experiment with the D2 DA receptor agonist, only the 0.1 μg quinpirole increased significantly the step-through latency during the test trials: retention was significant compared to the controls even 2 weeks after conditioning. The D2 DA receptor antagonist sulpiride pretreatment proved that the effect was due to the agonist induced activation of the D2 DA receptors of the VP. The supplementary experiment demonstrated that short-term memory is formed after conditioning in the experimental animals, supporting that the agonist enhanced memory consolidation in the first two experiments. Our results show that the activation of the D2 DA receptors in the VP facilitates memory consolidation as well as memory-retention in inhibitory avoidance paradigm.

Keeping the internal environment constant in a continuously changing external environment, i.e. the maintenance of homeostasis is crucial for the living organisms and it is served by several mechan...

The anterior cingulate cortex (ACC), is known to be intimately involved in food-related motivational processes and their behavioral organization, primarily by evaluating hedonic properties of the relevant stimuli. In the present study, the involvement of cingulate cortical interleukin-1β (IL-1β) mediated mechanisms in a) gustation associated facial and somato-motor behavioral patterns of Wistar rats were examined in taste reactivity test (TR). In addition, b) conditioned taste aversion (CTA) paradigm was performed to investigate the role of these cytokine mechanisms in taste sensation associated learning processes, c) the general locomotor activity of the animals was observed in open field test (OPF), and d) the potentially negative reinforcing effect of IL-1β was examined in conditioned place preference test (CPP). During the TR test, species specific behavioral patterns in response to the five basic tastes were analyzed. Response rates of ingestive and aversive patterns of the cytokine treated and the control groups differed significantly in case of the weaker bitter (QHCl, 0.03 mM), and the stronger umami (MSG, 0.5 M) tastes. IL-1β itself did not elicit CTA, it did not interfere with the acquisition of LiCl induced CTA, and it also failed to cause place preference or aversion in the CPP test. In the OPF paradigm, however, significant differences were found between the cytokine treated and the control groups in the rearing and grooming, the number of crossings, and in the distance moved. Our results indicate the involvement of cingulate cortical IL-1β mechanisms in the control of taste perception and other relevant behavioral processes.

Cognitive disturbances are among the most important features of schizophrenia, and have a significant role in the outcome of the disease. However, the treatment of cognitive symptoms is poorly effective. In order to develop new therapeutic opportunities, the MAM-E17 rat model of schizophrenia can be an appropriate implement. In the present study we investigated several cognitive capabilities of MAM-treated rats using radial arm maze (RAM) task, which corresponds to the recent research directives. Because of the diachronic appearance of schizophrenia symptoms and the early appearance of cognitive deficiencies, we carried out our experiments in three different age-periods of rats, i.e. in prepuberty, late puberty and adulthood. The performance of MAM-E17 rats was similar to control rats in the acquisition phase of RAM task, except for puberty. However, after rearrangement of reward positions (in the reverse paradigm) the number of errors of MAM-treated rats was higher in each age-period. In the reverse paradigm MAM-treated groups visited more frequently those non-rewarding arms, which were previously rewarding. Our results suggest that working memory of MAM-E17 rats is impaired. This deficit depends on the difficulty of the task and on the age-period. MAM-E17 rats seem to be more sensitive in puberty in comparison to controls. Diminished behavioral flexibility was shown as well. These behavioral results observed in MAM-E17 rats were similar to those of cognitive deficiencies in schizophrenia patients. Therefore, MAM-E17 model can be a useful implement for further research aiming to improve cognition in schizophrenia.

The ventromedial prefrontal cortex (vmPFC) of rats has reciprocal connections with the gustatory and the hedonic impact coding structures. The main goal of the present study was to investigate the involvement of local neurons of vmPFC and their catecholaminergic innervations in taste preference and taste reactivity test. Therefore, kainate or 6-hydroxydopamine (6-OHDA) lesions were performed in the vmPFC by iontophoretic method. In the first experiment, taste preference was tested to 250 mM and 500 mM glucose solutions over water in two-bottle choice test. In the second experiment, taste reactivity was examined to 4 concentrations of glucose solutions (250 mM, 500 mM, 750 mM and 1000 mM) and 4 concentrations of quinine solutions (0.125 mM, 0.25 mM, 1.25 mM and 2.5 mM). Our results showed, that kainate microlesion of vmPFC did not modify the preference of 250 mM and 500 mM glucose solutions in two-bottle choice test. In contrast, 6-OHDA microlesion of vmPFC resulted in increased preference to the higher concentration of glucose (500 mM) solution over water. Results of taste reactivity test showed that kainate lesion resulted in more ingestive and less rejective responses to 750 mM glucose solution and elevated rejectivity to the higher concentrations (1.25 mM and 2.5 mM) of quinine solutions. 6-OHDA lesion of vmPFC increased the number of ingestive responses to highly concentrated (500 mM, 750 mM and 1000 mM) glucose solutions and decreased the number of ingestive responses to the lower concentration (0.125 mM) of quinine solution. The present data provide evidence for the important role of vmPFC neurons and catecholaminergic innervation of the vmPFC in the regulation of hedonic evaluation of tastes and in the hedonic consummatory behavior.

The tachykinin NK1 receptor was suggested to be involved in psychiatric disorders, but its antagonists have failed to be effective as antidepressants in clinical trials. Hemokinin-1 (HK-1), the newest tachykinin, is present in several brain regions and activates the NK1 receptor similarly to substance P (SP), but acts also through other mechanisms. Therefore, we investigated the roles of the Tac4 gene-derived HK-1 in comparison with SP and neurokinin A (NKA) encoded by the Tac1 gene, as well as the NK1 receptor in anxiety and depression-like behaviors in mice. Mice lacking SP/NKA, HK-1 or the NK1 receptor (Tac1(-/-), Tac4(-/-), Tacr1(-/-), respectively) compared to C57Bl/6 wildtypes (WT), and treatment with the NK1 antagonist CP99994 were used in the experiments. Anxiety was evaluated in the light-dark box (LDB) and the elevated plus maze (EPM), locomotor activity in the open field (OFT) tests. Hedonic behavior was assessed in the sucrose preference test (SPT), depression-like behavior in the tail suspension (TST) and forced swim (FST) tests. FST-induced neuronal responsiveness was evaluated with Fos immunohistochemistry in several stress-related brain regions. In the LDB, Tac4(-/-) mice spent significantly less, while Tacr1(-/-) and CP99994-treated mice spent significantly more time in the lit compartment. In the EPM only Tac4(-/-) showed reduced time in the open arms, but no difference was observed in any other groups. In the OFT Tac4(-/-) mice showed significantly reduced, while Tac1(-/-) and Tacr1(-/-) animals increased motility than the WTs, but CP99994 had no effect. NK1(-/-) consumed markedly more, while Tac4(-/-) less sucrose solution compared to WTs. In the TST and FST, Tac4(-/-) mice showed significantly increased immobility. However, depression-like behavior was decreased both in cases of genetic deletion and pharmacological blockade of the NK1 receptor. FST-induced neuronal activation in different nuclei involved in behavioral and neuroendocrine stress responses was significantly reduced in the brain of Tac4(-/-) mice. Our results provide the first evidence for an anxiolytic and anti-depressant-like actions of HR-1 through a presently unknown target-mediated mechanism. Identification of its receptor and/or signaling pathways might open new perspectives for anxiolytic and anti-depressant therapies. 2016 Elsevier Inc. All rights reserved.

The role of dopamine (DA) receptors in spatial memory consolidation has been demonstrated in numerous brain regions, among others in the nucleus accumbens which innervates the ventral pallidum (VP). The VP contains both D1 and D2 DA receptors. We have recently shown that the VP D1 DA receptor activation facilitates consolidation of spatial memory in Morris water maze test. In the present study, the role of VP D2 DA receptors was investigated in the same paradigm. In the first experiment, the D2 DA receptor agonist quinpirole was administered into the VP of male Wistar rats in three doses (0.1, 1.0 or 5.0μg, respectively in 0.4μl physiological saline). In the second experiment, the D2 DA receptor antagonist sulpiride was applied to elucidate whether it can antagonise the effects of quinpirole. The antagonist (4.0μg, dissolved in 0.4μl physiological saline) was microinjected into the VP either by itself or prior to 1.0μg agonist treatment. Control animals received saline in both experiments. The two higher doses (1.0 and 5.0μg) of the agonist accelerated memory consolidation relative to controls and increased the stability of the consolidated memory against extinction. Sulpiride pretreatment antagonised the effects of quinpirole. In addition, the antagonist microinjected into the VP immediately after the second conditioning trial impaired learning functions. The present data provide evidences for the important role of VP D2 DA receptors in the consolidation and stabilization of spatial memory.

Neuropeptide oxytocin (OT) is involved in the regulation of social and non-social behaviour. The central nucleus of amygdala (CeA), part of the limbic system, plays an important role in learning, memory, anxiety and reinforcing mechanisms. CeA has been shown to be rich in OT receptors in rodents. Our previous findings indicated that OT in the rat CeA has a dose dependent rewarding and anxiolytic effect. The aim of our present study was to examine in the CeA the possible interaction of OT and D2 dopamine (DA) receptor antagonist Sulpiride on reinforcement in place preference test and on anxiety in elevated plus maze test. Wistar rats were microinjected bilaterally with 10 ng OT. In different group of animals 4 μg D2 DA receptor antagonist was applied. Other animals received D2 DA receptor antagonist 15 min before 10 ng OT treatment or vehicle solution into the CeA. Rats receiving 10 ng OT spent significantly longer time in the treatment quadrant during the test session in conditioned place preference test. Prior treatment with D2 DA receptor antagonist blocked the rewarding effects of OT. Antagonist in itself did not influence the time rats spent in the treatment quadrant. In elevated plus maze test, rats receiving 10 ng OT spent significantly longer time on the open arms. Prior treatment with D2 DA receptor antagonist blocked the effects of OT. Our results show that DA system plays a role in positive reinforcing and anxiolytic effects of OT because D2 DA receptor antagonist can block these actions.

The anterior cingulate cortex (ACC) is interrelated to limbic structures, parts of the central glucose-monitoring (GM) network. GM neurons, postulated to exist here, are hypothesised to participate in regulatory functions, such as the central control of feeding and metabolism. In the present experiments, GM neurons were identified and examined in the ACC by means of the multibarreled microelectrophoretic technique. After bilateral ACC microinjection of streptozotocin (STZ), glucose tolerance tests (GTTs), and determination of relevant plasma metabolite concentrations were performed. Body weights were measured at regular time points during the GTT experiment. Ten percent of the neurons - 30 of 282 recorded cells - responded to the administration of D-glucose, thus, declared to be the GM units. The peak values and dynamics of the GTT blood glucose curves, the plasma metabolite concentrations, and the weight gain were pathologically altered in the STZ treated animals. Our recording experiments revealed the existence of GM neurons in the anterior cingulate cortex. STZ induced selective destruction of these chemosensory cells resulted in feeding and metabolic alterations. The present findings indicate distinguished significance of the cingulate cortical GM neurons in adaptive processes of maintenance of the homeostatic balance.

Neurotensin (NT) acts as a neurotransmitter and neuromodulator in the central nervous system. NT is involved in reward and memory processes, drug addiction and also in the regulation of anxiety. The ventral pallidum (VP) receives neurotensinergic innervation from the ventral striatopallidal pathway originating from the nucleus accumbens. Positive reinforcing effects of NT in the VP had been shown recently, however the possible effects of NT on anxiety have not been examined yet. In our present experiments, the effects of NT on anxiety were investigated in the VP. In male Wistar rats bilateral microinjections of 100 ng or 250 ng NT were delivered in the volume of 0.4 μl into the VP, and elevated plus maze (EPM) test was performed. In another groups of animals, 35 ng NT-receptor 1 (NTR1) antagonist SR 48,692 was applied by itself, or microinjected 15 min before 100 ng NT treatment. Open field test (OPF) was also conducted. The 100 ng dose of NT had anxiolytic effect, but the 250 ng NT did not influence anxiety. The antagonist pretreatment inhibited the effect of NT, while the antagonist itself had no effect. In the OPF test there was no difference among the groups. Our present results show that microinjection of NT into the VP induces anxiolytic effect, which is specific to the NTR1 receptors because it can be eliminated by a specific NTR1 antagonist. It is also substantiated that neither the NT, nor the NTR1 antagonist in the VP influences locomotor activity.

In the present experiments, medial prefrontal cortical (mPFC) neurons were extracellularly recorded by means of tetrode electrodes to examine their possible role in the prediction of appetitive reward. Two different cue tones (CS) and sucrose solution or water reward (US) were associated in a Pavlovian conditioning paradigm. In order to test behavioral correlate of the CS-US association, the head acceleration before the first lick of licking cluster was measured. Neuronal activity changes in the mPFC were analyzed (i) during the CS presentations; (ii) before the first lick of licking clusters; (iii) during consummation; and (iv) we also examined whether consummation was represented in neurons responding to the CSs. There was a difference between the head accelerations to the different USs during early or late occurring first approaches, but there was no such a difference during intercluster approaches. A significant proportion of neurons changed their firing rate during the CS presentation, before the first lick of licking cluster or during licking of the reward. Both, excitatory and inhibitory responses were observed. A subpopulation of neurons responding to the CSs also responded during reward consumption. Differential population activities of excitatory neurons were recorded in response to the different CSs, CS evoked approach behaviors and consumption of different rewards. Neuronal responses also discriminated among the CSs and trials with or without consummation. These results provided evidence for the involvement of mPFC neurons in the prediction, representation and organization of conditioned behavioral actions, such as approaches to rewards and consummation.

Effects of 6-hydroxydopamine (6-OHDA) lesions in the medial prefrontal cortex with or without protection of norepinephrine (NE) fibers were examined on basic regulatory processes of feeding. Daily body weight, food and water intake were measured. Locomotor activity, ingestion after food or water deprivation, and preference for 5% and 10% glucose solution were examined. Dopamine (DA) and NE content, as well as, tyrosine hydroxylase immunoreactivity were assessed to confirm the neurotoxic effect of treatments. 6-OHDA lesions of the medial prefrontal cortex with or without NE fiber protection resulted in body weight loss. Diminished habituation in open field tests, i.e. a persistently high motor activity, was also observed. Application of 6-OHDA with NE fiber protection led to increased food consumption following food-deprivation and to enhanced glucose preference. Enhanced intake of 10% over 5% glucose solution was also detected. 6-OHDA lesion resulted in a decrease to 20% of NE tissue concentration and only to 75% of DA concentration. In case of lesion with NE protection the NE content decreased to 69% and DA level to 51% with significant loss of tyrosine hydroxylase positive fibers in the deeper layers of the medial prefrontal cortex. DA depletion in the medial prefrontal cortex resulted in increased behavioral responsiveness to hunger and glucose, as well as, to open field environment. Pronounced lesion of NE terminals caused increased reaction to the environment in open field but not to hunger or glucose solution.

The medial orbitofrontal cortex is involved in the regulation of feeding and metabolism. Little is known, however, about the role of local glucose-monitoring neurons in these processes, and our knowledge is also poor about characteristics of these cells.The functional significance of these chemosensory neurons was to be elucidated.Electrophysiology, by the multibarreled microelectrophoretic technique, and metabolic investigations, after streptozotocin induced selective destruction of the chemosensory neurons, were employed.Fifteen percent of the neurons responded to glucose, and these chemosensory cells displayed differential neurotransmitter and taste sensitivities. In acute glucose tolerance test, at the 30th and 60th minutes, blood glucose level in the streptozotocin-treated rats was significantly higher than that in the controls. The plasma triglyceride concentrations were also higher in the streptozotocin-treated group.Glucose-monitoring neurons of the medial orbitofrontal cortex integrate internal and external environmental signals, and monitor metabolic processes, thus, are indispensable to maintain the healthy homeostasis. Orv Hetil. 2017; 158(18): 692-700.