Correlations of Expression Levels of Lung Cancer Marker Gene Eno2 and Genes of Carcinogenesis and Apoptosis in the Hypothalamus of Mice with Depression-Like Behavior.

We experimentally demonstrated that chronic social stress during the development of a depression-like state enhances lung metastasis and modifies the expression of many carcinogenesis- and apoptosis-related genes in the hypothalamus of mice, including genes involved in lung cancer pathogenesis in humans. Analysis of the expression of genes encoding the major clinical markers of lung cancer in the hypothalamus of mice with depression-like behavior revealed increased expression of the Eno2 gene encoding neuron-specific enolase, a blood marker of lung cancer progression in humans. It was shown that the expression of this gene in the hypothalamus correlated with the expression of many carcinogenesis- and apoptosis-related genes. The discovered phenomenon may have a fundamental significance and requires further studies.

[Changes in Expression of Genes Associated with Calcium Processes in the Hippocampus in Mice Exposed to Chronic Social Stress].

Whole-transcriptome data were used to study the changes in expression of genes coding proteins involved in the calcium regulation processes in the hippocampus of male mice with symptoms of depression caused by chronic social defeat stress. Cacna1g, Cacnb3, Camk1g, Camk2d, Camk2n2, Caly, Caln1, S100a16, and Slc24a4 genes were upregulated in the hippocampus of depressed mice compared to a control, while Cacna2d1, Cacng5, Grin2a, and Calm2 were downregulated. The greatest number of significant correlations was observed between the expression level of Calm2, which showed the highest transcriptional activity, and other differentially expressed genes. Calcium signaling in the hippocampus was assumed to be disrupted in mice exposed to chronic social defeat stress. The involvement of Calm2, Camk1g, Camk2d, and Camk2n2 genes in the process is discussed.

[Serotonergic genes in the development of anxiety/depression-like state and pathology of aggressive behavior in male mice: RNA-seq data].

In course of daily agonistic interactions, mice tend to stratify into those with chronic social defeats and those that repeatedly display aggression, which lead to the development of mixed anxiety/depression-like state and the pathology of aggressive behavior, respectively. Using the data of whole transcriptome analysis (RNA-seq), the changes in the expression of serotonergic genes involved in the synthesis, inactivation, and reception of serotonin, as well as of the Creb1 (transcription factor) gene and the Bdnf (brain-derived neurotrophic factor) gene were detected in the striatum (STR), ventral tegmental area (VTA), midbrain raphe nuclei (MRN), hypothalamus (HYP), and hippocampus (HIP) of defeated and aggressive male mice. In mice of both groups, the Tph2, Ddc, Slc6a4, Htr2a, Htr3a, Htr5b, Slc18a2, and Bdnf genes were downregulated in the MRN and the Tph2, Ddc, and Slc6a4 genes were upregulated in the VTA. These changes were more significant in defeated mice. The Htr5b gene has first been shown to be involved in mechanisms of depression and pathology of aggressive behavior. In the defeated mice, the expression levels of the Htr4 and Aldh1b1 genes were increased in the MRN, and expression levels of the Maob, Htr4, Htr1a, and Slc18a2 genes were increased in the VTA, while the expression level of the Htr3a gene was decreased. In the HYP of aggressive mice the Maoa, Htr2a, Htr2c, and Creb1 genes were downregulated and the Htr6 gene was upregulated. In the defeated mice, the Maoa and Creb1 genes were downregulated and the Htr6 and Aldh1b1 genes were upregulated in the HYP. In the STR, the Htr1a gene was downregulated and the Htr7 and Bdnf genes were upregulated. The Htr1b gene was upregulated in the HIP. The coexpression of dopaminergic and serotonergic genes in the MRN and VTA in the control of pathological behaviors is discussed. Thus, the complex pattern of differential expression of serotonergic genes in brain regions developing under repeated agonistic interactions in mice in dependence on behavioral pathology have been observed.

[Altered Expression of Neurotransmitters Systems' Genes in the Ventral Tegmental Area of Depressive Male Mice: Data of RNA-Seq].

Chronic social defeat stress (CSDS) leads to the development of mixed anxiety/depression-like state in male mice similar to those in humans. It has been shown that, under CSDS, the adult brain undergoes changes in the functioning neurotransmitter systems in different brain regions. In this experiment we are focused on the analysis of expression of genes encoding proteins related with the metabolism and receptors of serotonin, catecholamines, GABA and glutamate in the ventral teg- mental area which is important for regulation of motivations, emotions and is involved into mech- anisms of affective disorders. Mixed anxiety/depression-like state was generated in male mice by exposure to CSDS during 20 days. The collected samples of the ventral tegmental area were se-- quenced at JSC Genoanalytica,(http://genoanalytica.ru/, Moscow, Russia).'We found that genes, related with serotonin (Tph2, Maob, SIc6a4, Htr4, Htr1a) were upregulated but expression of Htr3a gene was downregulated in the ventral tegmental area of depressive mice in comparison with the control. Besides, upregulation of dopaminergic Th, Ddc, Slc6a3, Sic18a2, Drd2, and Maob genes was found while noradrenergic Dbh, Slc6a2, Adra2c, and Adra2a genes were downregulated. Ex- pression of GABAergic Gabral, Gabra2, Gabrg2, Gabrg], Gabrq, Gad], and Gad genes as well as glutamatergic Grial, Gria2, Grik2, Grm2, Grm5, and Slc 7a8 genes were increased under CSDS. Development of mixed anxiety/depression-like state under CSDS in male mice is accompanied by increased expression of genes coding the proteins participating in the metabolism and receptions of serotonergic, dopaminergic, glutamatergic and GABAergic systems. Expression of genes coding the adrenergic reception is decreased. It is supposed that Drd2 H Htr3a genes may play the key role in the synchronization of other genes of neurotransmitter systems.

Role of Proteolytic Enzymes in the Interaction of Phytopathogenic Microorganisms with Plants.

Various forms of participation of proteolytic enzymes in pathogenesis and defense in plants are reviewed. Along with extracellular proteinases, phytopathogenic microorganisms produce specific effectors having proteolytic activity and capable of acting on proteins inside plant cells. In turn, for defense against pathogens, plants use both extracellular and intracellular proteinases.

[Changes in the Expression of Dopaminergic Genes in Brain Structures of Male Mice Exposed to Chronic Social Defeat Stress: An RNA-seq Study].

Whole-transcriptome analysis (RNA-seq) has been used to analyze changes in the expression of dopaminergic genes that encode proteins involved in the synthesis, inactivation, and neurotransmission of dopamine in the striatum, ventral tegmental area, raphe nuclei of the midbrain, hypothalamus, and hippocampus of male mice subjected to chronic social defeat. The expression of Th, Ddc, and Slc6A3 (Dat1) was upregulated, while that of Ppp1r1b and Sncg was downregulated in the ventral tegmental area; the expression of Th, Ddc, Drd2, and Sncg was downregulated in the raphe nuclei of midbrain; the expression of Th, Aldh2, and Ppp1r1b was upregulated, while that of Маоа was downregulated in the hypothalamus; Drd1 and Snca expression was downregulated and that of Sncb was upregulated in the striatum, and Sncb expression was upregulated in the hippocampus. There were no statistically significant changes in the expression of Comt, Maob, Drd3, Drd4, or Drd5 in the brain areas analyzed in stressed male mice (compared to control animals). Thus, the number of differentially expressed dopaminergic genes and the direction of expression changes in male mice experiencing chronic stress are specific to regions of the brain.

[Resistance of DBA/2J Mice to the Chronic Social Defeat Stress].

We investigated behavioral changes in male mice DBA/2J after the acquisition of a long experience of social defeats in agonistic interactions with aggressive partners of C57BL/6J and DBA/2J lines. The long experience of social defeat in DBA/2J mice did not change the strategy of theirbehavi6r during agonistic interactions. Reduced communicativeness and increased level of anxiety were found in the "partition" and "elevated plus maze" tests. There were no changes in locomotor activity in the "open field" test. After 20-30 days of social defeat stress there were no signs of depression, determined by the behavior during confrontations and in the Porsolt test. There was no sign of catalepsy, decreased exploratory be- havior and impaired social recognition. Thus, the mice of this strain can be considered relatively resis- tant to the development of depressive-like state under chronic social stress and may be used for the study of the mechanisms of such stability.

[Relationship of Anxiety and Depression in the Development of Mixed Anxiety/Depression Disorder. An Experimental Study of Comorbidity Mechanisms (Review)].

As clinical practice and experimental studies show, symptoms of depression and anxiety often accompany each other. It is well known that combination of anxiety and depression in patients is treated more slowly, requires large doses of drugs, increases the likelihood of suicide and often leads to relapse. Furthermore, antidepressants and anxiolytics exert its therapeutic effect in limited cases even in monopolar anxiety or depression state. In this review of literature and our own data the relationship of anxiety and depression is analyzed. It has been shown with using the model of mixed anxiety/depression disorder caused by chronic social defeat stress, that the anxiety and depression are changed under the influence of psychotropic drugs independently.

[Serotonergic Control of Aggressive Behavior: Novel Approaches--New Interpretations (Review)].

The concept of a major inhibitory role of serotonin in aggressive behavior is widely accepted by researchers. There were ample evidences that a pharmacologically-induced increase in serotonergic activity attenuates agonistic behavior, and the manipulations inhibiting the brain serotonergic system can elicit aggressiveness in animals. Ealier, experimental studies have demonstrated reduced metabolism in brain serotonergic system and specific changes in pharmacological sensitivity of 5-HT(1A) receptors in male mice with repeated experience of aggression. It has been shown that mRNA levels of the serotonergic genes in the midbrain raphe nuclei are reduced in aggressive males. After no-fight period expression of some genes is restored or becomes higher compared with the controls on the background of increased aggression. The review provides data supporting and contradicting the serotonin-deficiency hypothesis of increased aggressiveness, revealing non-specific role of serotonin in the control of aggressive behavior, and also provides information about the inhibition of serotonergic activity under repeated experience of aggression.

[Alteration of Social Behaviors in Male Mice of CBA/Lac Strain under Agonistic Interactions].

Ability of people to communicate with each other is a necessary component of social behavior and normal development of individuals living in community. A pronounced impairment in communication may be the result of autism which is characterized by impaired socialization, low communication and restricted and/or repetitive behaviors. It is hypothesized that genes or rare mutations play a key role in the development of autism. However a multifold increase of the cases with autistic spectrum symptoms over the last years cannot be attributed exclusively to genetic mutations or heredity. Environmental contribution to the development of autistic symptoms has to be considered. The paper aimed to analyze the social behaviors of CBA/Lac mice with repeated experience of aggression or social defeats in daily agonistic interactions with accent on searches of associations with autistic symptoms in comparison with previously studied C57BL/6J animals. It has been shown that male mice of both strains with alternative social behaviors demonstrated the changes in social behaviors; however the expression of some form of behaviors was different. The data obtained to assert that long-term hostile social environment lead to development of disturbances in social behaviors, accompanying by autistic-like symptoms.

[Anxiogenic and anxiolytic effects of lithium chloride under preventive and therapeutic treatments of male mice with repeated experience of aggression].

Repeated experience of aggression in daily agonistic interactions is accompanied by development of changes in behaviors and psychoemotional states indicating the development of the psychopathology of aggressive behavior, which are difficult to correct by drugs used for decrease of aggression in the clinics. In this paper the influence of lithium chloride on the behavior of aggressive males in different tests assessing anxiety, communication and exploratory activity (elevated plus maze test, social interaction test, partition test), as well as aggressiveness (agonistic interaction test) were studied. Lithium chloride (Sigma-Aldrich Co, 100 mg/kg/day, i.p.) was administered preventively to male in ranging from the 7th day of agonistic interactions, as well as therapeutically to males with 21 days of aggression experience during the period without agonistic interactions. Also the effects of chronic lithium chloride treatment on behaviors of animals without agonistic interactions (intact mice) were studied. Period of drug and saline (as the controls) treatment--14 days. It has been shown that preventive lithium chloride treatment of male mice with repeated experience of aggression induced pronounced anxiogenic effect, under therapeutic treatment--nxiolytic effects. Anxiolytic effect was also observed in intact males. There is no effect of lithium chloride on aggression. Differences in the effects of lithium chloride under preveitive and therapeutic treatments, as well as the causes of individual sensitivity to the drug in male mice in one group were discussed.

Cell cycle arrest in the thymus and spleen in male mice under conditions of chronic social defeat stress: effects of diazepam.

The effects of chronic social defeat stress on the percentage of cells in different phases of the cell cycle and in apoptosis in the thymus and spleen of male mice were studied by the method of flow cytofluorometry. In stressed males, thymus weight decreased, the percent of proliferating thymocytes was significantly lower, and the percentage of G0-G1 cells was higher than in intact males. Stress substantially reduced the percentage of splenocytes in the G0-G1 phase and apoptotic cells, but the percentage of S and G2-M cells and proliferation index significantly increased. Chronic administration of anxiolytic diazepam prevented the majority of the changes in the percentage of cells in different phases of the cell cycle, but apoptosis in the thymus increased under these conditions. Possible association between cell cycle disorders, impairment of cell immunity, and chronic anxiety developing under conditions of long-term social defeat stress is considered.

An experimental approach for the study of psychotropic drug effects under simulated clinical conditions.

The sensory contact model can induce various different psychopathological states in male mice (anxious depression, catalepsy, social withdrawal, pathological aggression, cognition disturbances, anhedonia, alcoholism etc.). Additionally, this model facilitates the screening of drugs for therapeutic properties, preventive properties and efficiency under simulated clinical conditions. This approach can reveal the action of drugs at different stages of disease development. It is proposed that this pharmacological approach can be applied for the screening of various novel psychotropic drugs.

The effects of the D1 receptor antagonist SCH-23390 on individual and aggressive behavior in male mice with different experience of aggression.

The dopaminergic systems of the brain are known to be involved in the mechanisms of aggression. The present report describes studies of the effects of acute administration of the dopamine D1 receptor antagonist SCH-23390 (0.1 mg/kg, i.p., over 30 min) on the individual and aggressive behavior of male mice of the line C57BL/6J with different experience of aggression. A group of animals with no previous experience of aggression responded to administration of this agent with decreases in direct aggression (attacks), though the total time of hostile behavior, i.e., the sum of the durations of attacks, aggressive grooming, and scattering of foreign litter, showed no change. The agent had no effect on the aggressivity of animals with 20 days of experience of agonistic confrontations. The discussion addresses the possible development of pharmacological densensitization of dopamine D1 receptors in aggressive males in response to prolonged activation of the dopaminergic systems in conditions of repeated experience of aggression, as demonstrated previously.

A model of anxious depression: persistence of behavioral pathology.

Chronic psychoemotional stress induced by negative experience of social defeats in intermale confrontations over a period of 30 days was found to lead to the development of anxious-depressive symptomatology in male mice. Cessation of the psychopathogenic conditions and placing of depressed animals in comfortable conditions for 1-2 weeks with females did not lift the pathological state. Individuals continued to show marked anxiety, a behavioral deficit, decreased communicativeness, and a high level of depressivity, as revealed by a variety of behavioral tests. Persistence of the resulting psychoemotional disturbance in these animals is evidence for the development and persistence of the behavioral pathology requiring drug treatment.

Effect of repeated experience of victory and defeat in daily agonistic confrontations on brain tryptophan hydroxylase activity.

The rate-limiting enzyme of serotonin biosynthesis, tryptophan hydroxylase (TPH), was studied in brain areas of male mice with repeated experience of victory (winners) or defeat (losers) gained in 10 daily agonistic confrontations. A reduction of TPH activity in the midbrain and an increase in the hypothalamus was demonstrated for winners compared with controls. In contrast, repeated defeat in social confrontations was associated with higher TPH activity in the striatum and hypothalamus in losers compared with controls. Agonistic interactions did not affect TPH activity in the amygdala, nucleus accumbens or hippocampus in either winners or losers. The sensory contact technique used in this work for generating winners and losers may be productive in the analysis of TPH gene regulation.

Increase of tyrosine hydroxylase and dopamine transporter mRNA levels in ventral tegmental area of male mice under influence of repeated aggression experience.

Tyrosine hydroxylase (TH) and dopamine transporter (DAT) mRNA levels in the ventral tegmental area (VTA) of midbrain were measured by multiplex RT-PCR in male mice with repeated experience of social victories (winners) and social defeats (losers) in 10 daily agonistic confrontations. Two independent experiments revealed enhanced TH and DAT mRNA levels in VTA of the winners in comparison with the losers and controls (animals after 5 days of individual housing). A positive correlation between DAT and TH mRNA levels was shown.

Chemical characterization of effective and ineffective strains of Rhizobium leguminosarum bv. viciae.

Chemical composition of lipopolysaccharide (LPS) isolated from an effective (97) and ineffective (87) strains of R. l. viciae has been determined. LPS preparations from the two strains contained: glucose, galactose, mannose, fucose, arabinose, heptose, glucosamine, galactosamine, quinovosamine, and 3-N-methyl-3,6-dideoxyhexose, as well as glucuronic, galacturonic and 3-deoxyoctulosonic acid. The following fatty acids were identified: 3-OH 14:0, 3-OH 15:0, 3-OH 16:0, 3-OH 18:0 and 27-OH 28:0. The ratio of 3-OH 14:0 to other major fatty acids in LPS 87 was higher that in LPS 97. SDS/PAGE profiles of LPS indicated that, in lipopolysaccharides, relative content of S form LPS I to that of lower molecular mass (LPS II) was much higher in the effective strain 97 than in 87. All types of polysaccharides exo-, capsular-, lipo, (EPS, CPS, LPS, respectively) examined possessed the ability to bind faba bean lectin. The degree of affinity of the host lectin to LPS 87 was half that to LPS 97. Fatty acids (FA) composition from bacteroids and peribacteroid membrane (PBM) was determined. Palmitic, stearic and hexadecenoic acids were common components found in both strains. There was a high content of unsaturated fatty acids in bacteroids as well as in PBM lipids. The unsaturation index in the PBM formed by strain 87 was lower than in the case of strain 97. Higher ratio of 16:0 to 18:1 fatty acids was characteristic for PMB of the ineffective strain.

Social success and voluntary ethanol consumption in mice of C57BL/6J and CBA/Lac strains.

Voluntary ethanol (20% solution) consumption in mice of C57BL/6J (C57) and CBA/Lac (CBA) strains with consecutive experience of victories (winners) or defeats (losers) in daily intermale agonistic confrontations were studied. Winners and losers of the CBA strain maintained the ethanol consumption on the same level. Losers of the C57 strain increased the ethanol consumption most dramatically during the second week of testing, while winners of this strain did not change the ethanol intake. The aspiration to the social contact estimated by behavioral activity as a reaction to the other male significantly increased in "drinking" losers of the C57 strain. The influence of heredity and emotional state of animals formed by their social success on voluntary ethanol consumption are discussed.

Comparative analysis of anxiety-like behavior in partition and plus-maze tests after agonistic interactions in mice.

The behavior of C57BL/6J male mice with experience of repeated victories (winners) or defeats (losers) in daily agonistic interactions, was examined in the plus-maze and partition tests. The latter procedure assesses the reactivity of mice to another conspecific in the neighboring compartment of a common cage, communicative behavior or level of sociability. The behavior of mice after 10 days (T10) and 20 days (T20) of agonistic confrontations, as well as in controls (5 days of individual housing) was analyzed. Significant differences were found between T10 and T20 losers and controls in both tests. In the partition test, a decrease in the number of approaches and total and average time spent near the partition was found in T10 and, more pronounced, in T20 losers. In the plus-maze, losers showed fewer open and total entries than controls. Moreover, rarely did they pass from one enclosed arm to another, and also showed a decreased number of 'peepings' from enclosed arms. Percentage of open time did not differ significantly in losers compared to controls. It is suggested that the level of losers' sociability, estimated as low by the partition test, can include anxiety as a component, which is confirmed, at least partly, by some parameters of the plus-maze test. There were no differences in the partition or plus-maze tests between T10 winners and controls. However, all parameters of partition behavior were significantly different between T20 winners and controls. In the plus-maze, similar to losers, T20 winners displayed fewer open arm entries, number of "peepings,' and passages than controls. It was concluded that parameters of the partition and plus-maze tests correlate in mice with the alternative experience of agonistic confrontations, positive or negative. A combination of these two tests may be used for estimation of developing anxiety in mice.