[The effect of prenatal stress on antioxidant glutathion-assosiated enzymes activity in subcellular fractions of rat's liver]. / Vliianie prenatal'nogo stressa na aktivnost' glutation-zavisimykh antioksidantnykh fermentov v subkletochnykh fraktsiiakh pecheni krys.

The activity of glutathione-associated antioxidant enzymes in subcellular fractions (cytosolic, mitochondrial, and cell nucleus fractions) was investigated in the liver of adult male Wistar rats born after prenatal stress was. Two groups of animals were studied in the experiment: (1) control group included - animals was born by intact mothers, and (2) prenatal stress group included animals whose mothers were subjected to immobilization stress in high-light conditions from the 15th to the 19th day of pregnancy. The activity of glutathione peroxidase (EC 1.11.1.9) in prenatally stressed animals decreased in the fractions of nuclei and mitochondria compared to the control group, while the activity of glutathione reductase (EC 1.8.1.7.) increased in the same subcellular fractions. The activity of glutathione transferase (EC 2.5.1.18) in prenatally stressed rats reduced in the cytosol and mitochondrial fractions as compared to control group. Redistribution of the antioxidant enzyme activity in the cytosol, the fraction of nuclei and the mitochondrial fraction of liver tissue may contribute to the formation of the pathological phenotype of prenatally stressed offspring.

Behavior Disorders Caused by Perinatal Hypoxia in Juvenile Rats and Their Correction with GABA Derivative.

We studied the effects of acute normobaric hypoxia on postnatal day 2 (model of preterm pregnancy) on reflex activity and behavior of juvenile male Wistar rats and the possibility of correction of behavioral deficit by administration of GABA derivative Salifen after hypoxia. It is shown, that perinatal hypoxia impaired righting reflex and forelimb grip strength and increased motor activity in juvenile male rats. Administration of Salifen for 14 days in a dose of 15 mg/kg improved reflex activity and behavior of rats, which indicates the prospect of further study of the therapeutic efficacy of this drug on models of neonatal encephalopathy.

[ACTIVITY OF HYPOTHALAMIC-PITUITARY-ADRENAL AXIS OF PRENATALLY STRESSED MALE RATS IN EXPERIMENTAL MODEL OF DEPRESSION].

The hypothalamic-pituitary-adrenal (HPA) axis activity changes were examined in the adult, prenatally stressed male rats in the experimental depression model--the paradigm of "learned helplessness". It was shown that in males descending from intact mothers a depressive-like state was accompanied by an increase in HPA activity. The expression of corticotrophin-releasing hormone (CRH) in the hypothalamic paraventricular nucleus (PVN) increases, coupled with a rise in plasma levels of ACTH and corticosterone as well as in adrenal weight. At the same time in males born from mothers stressed during the last week of pregnancy we observed a decrease in activity of both the central (hypothalamus) and the peripheral (adrenal cortex) parts of regulation of this hormonal axis similar to that revealed for these animals in our previous study in "stress-restress" paradigm. It is concluded that prenatal stress modifies the sensitivity of animals to the inescapable intense stress impact, which manifests itself in a specific pattern of the HPA axis activity after stress load.

Activity of the Hypothalamic-Pituitary-Adrenal System in Prenatally Stressed Male Rats on the Experimental Model of Post-Traumatic Stress Disorder.

Using the experimental model of post-traumatic stress disorder (stress-restress paradigm), we studied the dynamics of activity of the hypothalamic-pituitary-adrenal system (HPAS) in adult male rats, whose mothers were daily subjected to restraint stress on days 15-19 of pregnancy. Prenatally stressed males that were subjected to combined stress and subsequent restress exhibited not only increased sensitivity of HPAS to negative feedback signals (manifested under restress conditions), but also enhanced stress system reactivity. These changes persisted to the 30th day after restress. Under basal conditions, the number of cells in the hypothalamic paraventricular nucleus of these animals expressing corticotropin-releasing hormone and vasopressin was shown to decrease progressively on days 1-30. By contrast, combined stress and restress in control animals were followed by an increase in the count of CRH-immunopositive cells in the magnocellular and parvocellular parts of the paraventricular nucleus and number of vasopressin-immunopositive cells in the magnocellular part of the nucleus (to the 10th day after restress). Our results indicate a peculiar level of functional activity of HPAS in prenatally stressed males in the stress-restress paradigm: decreased activity under basal conditions and enhanced reactivity during stress.

[STRESS-INDUCED PATTERNS OF THE HYPOTHALAMIC CRH AND VASOPRESSIN EXPRESSION IN FEMALE RATS IN A MODEL OF POSTTRAUMATIC STRESS DISORDER].

The neuroendocrine mechanisms underlying anxiety-like state development in cycling female rats with different plasma estradiol levels have been studied in a stress-restress paradigm, an animal model of posttraumatic stress disorder (PTSD). The effect of stress-restress on the hypothalamic expression of corticotropin-releasing hormone (CRH) and vasopressin was analyzed by quantitative immunocytochemistry. Stress-restress was found to increase CRH expression in the hypothalamic paraventricular nucleus (PVN) on the 10th post-restress day, but the level of CRH expression in the PVN restored to the basal values on the 30th post-restress day in all experimental groups. It was shown an increase in vasopressin immunoreactivity in the PVN from the 10th to the 30th post-restress days in female rats exposed to stress during the estrus phase (low plasma estradiol level). In summary, female rats with low plasma estradiol level exhibited the most significant changes in the hypothalamic neuroendocrine system following stress-restress exposure. It might be hypothesized that hyperactivity of the hypothalamic circuit of the central vasopressinergic system is one of the possible mechanisms underlying PTSD-like state development in female rats in a stress-restress paradigm.

Development of behavioral and hormonal disorders in prenatally stressed female rats on the model of posttraumatic stress disorder.

The dynamics of changes in behavioral and hormonal manifestations of a pathological state in mature female rats born by mothers exposed to daily restraint stress on days 15-19 of pregnancy were studied in the experimental model of posttraumatic stress disorder (stress-restress paradigm). Experiments demonstrated increased anxiety in control and prenatally stressed female rats after combined stress followed by restress. This parameter remained enhanced until day 10 after restress in control rats and day 30 in prenatally stressed animals. The severity of depression increased on days 1 and 10 after restress in prenatally stressed female rats. Basal activity of the pituitary-adrenocortical axis increased only in prenatally stressed female rats under these conditions. This parameter increased 1 day after restress and decreased after day 30. It was concluded that prenatal stress could increase the predisposition to post-stress mental pathologies in experimental animals, which are manifested in increased severity and duration of behavioral and hormonal impairments.

[Modification of expression of neurohormones in hypothalamus of prenatally stressed male rats in model of posttraumatic stress disorder].

By the method of quantitative immunohistochemistry there has been studied expression of corticotrophin-releasing hormone (CRH) and vasopressin in hypothalamic paraventricular nucleus (PVN) of prenatally stressed rats in the experimental model of the posttraumatic stress disorder--the paradigm "stress-restress". The prenatal stress was modeled by immobilization of pregnant female rats for 1 h from the 15th to the 19th day of pregnancy. It has been shown that in sexually mature males--descendants of stressed mothers--a decrease in immunoreactivity to CRH and vasopressin is observed in the parvocellular and magnocellular PVN areas 10 days after the restress. In the control group males born by intact mothers the level of immunoreactivity to CRH was increased in both PVN areas, whereas with respect to vasopressin--in the magnocellular area. Only in the prenatally stressed males there is detected a decrease in the corticosterone level in the blood plasma 10 days after the restress. It is concluded that in the control group males themanifestation of the pathological state in the paradigm "stress-restress" consists in hyperactivation of the hypothalamic chain of regulation of the hypothalamo-pituitary-adrenocortical system, whereas in the prenatally stressed animals, on the contrary, there is observed a decrease in activity both of the central (PVN) and of the peripheral (adrenal cortex) chain of this hormonal axis.

Effects of prenatal stress on the activity of the pituitary-ovarian system in female rats.

The comparative analysis of hormonal status was performed in mature female rats with experimental deficiency of estrogens, which mothers were exposed to stress during pregnancy. High levels of follicle-stimulating hormone and luteinizing hormone, and significantly lower amount of estradiol were observed in intact prenatally stressed females in comparison with intact non-stressed female rats. The increase in the levels of follicle-stimulating hormone and luteinizing hormone, and the decrease in estradiol concentration were more pronounced in blood serum of prenatally stressed ovariectomized rats as distinct from intact non-stressed and prenatally stressed female rats, and non-stressed ovariectomized female rats. We can conclude that prenatally stressed ovariectomized rats were characterized by an increased sensitivity to exogenous hormonal interventions and high lability of functional state of the pituitary-ovarian system.

[Influence of fluoxetine and paroxetine on anxiety-like behavior in young and adult prenatally stressed male rats].

The aim of the present work was an estimation of effects of chronic administration of selective serotonin reuptake inhibitors--fluoxetine (5.0 mg/kg, p.o.) and paroxetine (5.0 mg/kg, p.o.) for 14 days of postnatal period on anxiety-like behavior in the prenatally stressed male rats during pubertal period (1,5 month) and the adult state (3 month). Chronic paroxetine administration to females failed to change an anxiety-like behavior independently from age. On the contrary, administration of fluoxetine resulted in modulating influence on the anxiety-like behavior of prenatally stressed rats dependently from age: anxiolytic effect was noted in young males, while anxiogenic effect was observed in the adult male rats.

Age-related peculiarities in steroid hormone secretion and behavior in prenatally stressed female rats in novel environment.

We studied the effects of immobilization of female rat during days 15-18 of pregnancy on the behavior in novel environment (open field test) and blood level of steroid hormones in their female offspring depending on the cycle phase and age. The rats were tested at the age of 3, 12, and 24 months. Locomotor and exploratory activity and anxiety of control rats depended on the phase of estrous cycle. Age-related changes in the studied parameters were noted: locomotor and exploratory activity decreased and anxiety increased against the background of reduced secretion of the main ovarian hormones with age. In addition, blood estradiol level and behavior in novel environment virtually did not depend on age and estrous cycle phase in prenatally stressed females. Our findings suggest that maternal stress has a modulatory effect on relationship between behavioral type and estrous cycle stage, as well as on age-related pattern of behavioral reactions.

Blockade of 5-HT2A/2C-type receptors impairs learning in female rats in the course of estrous cycle.

We studied the effects of chronic administration (14 days) of agonist of 5-HT2B/2C serotonin receptors m-CPP (0.5 mg/kg subcutaneously) and agonist of 5-HT2A/2C serotonin receptors ketanserin (0.1 mg/kg intraperitoneally) on conditioned reactions in female rats in different phases of the estrous cycle. Passive avoidance (PA) paradigm and Morris water maze were used as behavioral tests. Chronic administration of m-CPP did not affect PA retrieval during the proestrus and estrus phases, but improved the dynamics of spatial learning in Morris water maze in comparison with control rats. Chronic administration of ketanserin uniformly impaired processes of spatial and nonspatial learning in female rats irrespective to the phase of the estrous cycle. A modulating role of 5-HT2A/2C and 5-HT2B/2C serotonin receptors in process of learning in female rats during the key phases of the estrous cycle was demonstrated.

Effects of 8-OH-DPAT and NAN-190 on anxious-depressive-like behavior of female rats during the estrous cycle.

We studied the effect of chronic administration of 5-HT(1A)-receptor agonist 8-OH-DPAT (0.05 mg/kg subcutaneously) or antagonist NAN-190 (0.1 mg/kg intraperitoneally) for 14 days on anxious-depressive-like behavior of female rats during the key phases of the estrous cycle. Chronic administration of NAN-190 during the estrus phase produced an anxiogenic effect, while its administration during proestrus phase induced an anxiolytic effect. Administration of 8-OH-DPAT had no effect on anxiety level, but produced a pronounced antidepressive effect irrespective of the phase of the estrous cycle.

The possible use of hypoxic preconditioning for the prophylaxis of post-stress depressive episodes.

The protective effects of hypoxic preconditioning on the development of depressive states in rat models were studied. Three episodes of intermittent preconditioning using hypobaric hypoxia (360 mmHg, 2 h) prevented the onset of depressive behavioral reactions, hyperfunction of the hypophyseal-adrenal system, and impairments in its suppression in the dexamethasone test in rats following unavoidable aversive stress in a model of endogenous depression. The anxiolytic and antidepressant actions of hypoxic preconditioning in experiments on rats were no less marked than those of the tetracyclic antidepressant ludiomil. The results obtained here provide evidence that preconditioning with intermittent hypobaric hypoxia increases resistance to psychoemotional stresses, has marked anxiolytic and antidepressant effects, and can be used for the prophylaxis of depressive episodes.

Expression of glucocorticoid receptor in the brain of rats during postnatal ontogeny.

Expression of glucocorticoid receptor protein in the hippocampus and frontal cortex of male and female rats during the 1st, 2nd, and 3rd weeks of life was studied by Western blot hybridization. In the frontal cortex, the concentration of receptor protein progressively increased from the 1st to the 3rd week of life; in females, expression of 94-kDa protein significantly surpassed that in males during the 1st week of life. In the hippocampus, expression of 94-kDa and 82-kDa proteins during the 1st week of life was higher in males. Moreover, expression of the major glucocorticoid receptor isoform (94 kDa) in this structure remained unchanged in all periods of the study in males, whereas in females it was low over the first 2 weeks of life and increased by the 3rd week. Variations in the expression of glucocorticoid receptors in the hippocampus of male and female rats coincide with changes in plasma corticosterone concentration during the early postnatal ontogeny.

Involvement of nuclear progesterone receptors in the formation of anxiety in female mice.

We report here studies on the delayed effects of exogenous progesterone on the formation of anxiety in female mice. Ovariectomized female mice were given seven days of replacement therapy either with the two main ovarian hormones-progesterone and estradiol benzoate-or with progesterone only; levels of anxiety were measured six hours later in the elevated plus-maze. The role of nuclear progesterone receptors in controlling the level of anxiety was assessed by giving some mice injections of the synthetic progesterone receptor blocker mifepristone 2 h before the last dose of hormones. An immunohistochemical method was used to study changes in the number of nuclear progesterone receptors in different areas of the brains of experimental animals. These studies showed that progesterone has a delayed enhancing effect on anxiety in female mice. The role of nuclear progesterone receptors in forming this behavioral characteristic was supported by a strong correlation between changes in the numbers of progesterone receptor-immunopositive cells in several brain structures and the level of anxiety. Prior blockade of progesterone receptors using mifepristone led to a maximal reduction in the level of anxiety, which was also evidence for a role for the genomic mechanisms of action of progesterone in controlling anxiety in females.

Effects of impaired testosterone metabolism during prenatal ontogenesis on the level of anxiety and behavior of rats in a novel environment.

The effects of administration of the aromatase inhibitor 1,4,6-androstatrien-3.17-dione (ATD) to female rats during the last third of pregnancy on the formation of behavior of offspring of both genders in a novel environment were studied. Animal behavior was assessed in the open field and elevated cross maze tests. Inhibition of testosterone aromatization during the prenatal period of development resulted in increases in anxiety and emotionality in experimental rats at age one month; increases in these measures in adult animals were seen in both males and females exposed to prenatal ATD. Intergender differences between control males and experimental females, in terms of behavioral measures in the novel environment such as motor activity, the duration of the freezing and grooming reactions, as well as well the level of anxiety, disappeared. It is concluded that impairment of testosterone metabolism during the prenatal period of development affects the formation of the behavior of rats in a novel environment as determined by genetic gender.

Transcranial electrostimulation activates reparative regeneration and the insulin-producing function of pancreatic B-cells in alloxan diabetes in rats.

Studies on rats with experimental diabetes induced by administration of alloxan showed that transcranial electrostimulation of endorphinergic brain structures stimulates the regeneration of damaged beta-cells in pancreatic islets of Langerhans. This was identified on pancreatic sections stained with hematoxylin and eosin. De novo formation of small islets was noted, as evidenced by their regeneration from progenitor cells. After transcranial electrostimulation, islet beta-cells stained by the Gomori method showed recovery of granularity - a sign of insulin production. Application of an immunoenzyme method demonstrated recovery of blood insulin levels, the dynamics of increases in which showed a highly significant negative correlation with a decrease in blood glucose. These data led to the conclusion that the antihyperglycemic effect of transcranial electrostimulation in experimental alloxan diabetes results from reparative regeneration of beta-cells in islets of Langerhans with recovery of their insulin-producing function.

Characteristics of behavior and stress reactivity of the hypophyseal-adrenocortical system in rats with prenatal inhibition of testosterone metabolism.

The effects of administration of the aromatase blocker 1,4,6-androstatrien-3,17-dione (ATD) to female rats in the last third of pregnancy on the stress reactivity of the hypophyseal-adrenocortical system (HACS), behavior in a novel environment (an open field), and anxiety in an elevated cross maze in their adult offspring of both genders were studied. Inhibition of testosterone aromatization in the brain during the prenatal period of development was found to lead to a decrease in the basal activity of the HACS in males and longer-lasting hormonal stress responses in animals of both genders. However, the intergender differences in the nature of the stress reactivity of the system in the experimental animals persisted. Prenatal administration of ATD also induced increases in the levels of anxiety and emotionality and the duration of grooming reactions in males and females and eliminated intergender differences between control males and experimental females in terms of measures of behavior in a new environment such as movement activity, duration of the freezing reaction, and grooming. These data led to the conclusion that impaired testosterone metabolism in the brain during the prenatal period of development induced by administration of the aromatase blocker leads to changes in the nature of the stress response of the HACS in adult male and female rats and impairs the formation of sexual dimorphism in anxiety levels and the extent of behavioral reactions to environmental novelty in females.

Changed activity of the hypothalamic-pituitary-adrenocortical system in prenatally stressed female rat during aging.

We studied the effects of daily 1-h immobilization of female rats on days 15-18 of pregnancy on functional activity of the hypothalamic-pituitary-adrenocortical system and its sensitivity to regulatory signals realized by the negative feedback mechanism in female progeny during aging. Prenatal stress potentiated the inhibitory processes in young animals. In aging female rats, the sensitivity of the hypothalamic-pituitary-adrenocortical system to feedback signals significantly decreased and circadian stress reactivity was disturbed. These data suggest that maternal stress modifies the age-related pattern of the hypothalamic-pituitary-adrenocortical regulation in female progeny.