Melatonin modulate the expression of α1- and ß2-adrenoceptors in the hippocampus of rats subjected to unpredictable chronic mild stress.

OBJECTIVE: This study investigated the effects of chronic melatonin treatment on gene expression of α1-, α2-, ß1- and ß2-adrenoceptors in the hippocampus of rats subjected to chronic unpredictable mild stress (CUMS). BACKGROUND: Preclinical studies have also shown that melatonin prevented short- and long-term memory impairments and exhibited antidepressant-like actions. METHODS: For this study, we used 24 animals, which were divided into four groups, and the experiment lasted 4 weeks. We quantified the changes in mRNA and protein levels of α1-, α2-, ß1- and ß2-adrenoceptors in the hippocampus after melatonin treatment. RESULTS: Our results demonstrated a decreased gene expression of α1-, α2- and ß2-adrenoceptors in the hippocampus of rats subjected to unpredictable chronic mild stress, while there was no change in gene expression of ß1-adrenoceptors. Melatonin treatment in the CUMS rats prevented the stress-induced decrease in mRNA and protein levels of α1-and ß2-adrenoceptors, whereas did not affect either on mRNA or protein level of ß1-and α2-adrenoceptors. CONCLUSION: Our data suggest that melatonin, by increasing reduced levels of α1- and ß2-adrenoceptors mRNA and protein in the hippocampus of chronic stressed rats, may be beneficial in conditions such as chronic stress and provides an experimental opportunity to probe into further molecular mechanisms underlying the regulation of these receptor subtype (Fig. 2, Ref. 28).

Anxiety and Hyperlocomotion Induced by Chronic Unpredictable Mild Stress Can Be Moderated with Melatonin Treatment.

Preclinical studies have shown that melatonin exercised antidepressant-like and anxiolyticlike effects in animal models of anxiety. The aim of the present study was to correlate the changes in behaviour induced by melatonin treatment with the activity of the dopaminergic system in the hippocampus of Wistar rats exposed to chronic, unpredictable, mild stress (CUMS). Male Wistar rats, 11 weeks old, were subjected to chronic stress for 28 successive days. Separate groups of control and stressed rats were intraperitoneally injected daily either with melatonin (10 mg/kg/day, i.p.) or placebo (5% ethanol). The open-field and elevated plus-maze tests were used to assess locomotor activities and anxiety levels. The content of dopamine (DA) in the hippocampal tissues was determined using radioenzymatic assay, while changes in tyrosine hydroxylase (TH) mRNA and protein levels in the hippocampus were determined using real-time RT-PCR and Western immunoblotting. Chronic stress led to reduction in the hippocampal dopaminergic content without affecting the levels of TH protein. These changes were accompanied by increased locomotor activity and higher anxiety levels in the open-field test. Administration of melatonin for 28 days resulted in an increase in the hippocampal DA content as a result of elevated TH protein levels. Melatonin showed an improvement in anxiety-like behaviour along with significantly reduced exploration. We could conclude that melatonin may stimulate dopaminergic synthesis in the hippocampus in order to suppress stress-induced behaviour.

Differential expression of tyrosine hydroxylase and transporters in the right and left stellate ganglion of socially isolated rats.

Chronic isolation stress of adult rat males acted increasing gene expression of tyrosine hydroxylase (TH) and neuronal norepinephrine transporter (NET) in the right stellate ganglia, while vesicular monoamine transporter 2 (VMAT2) level remained unchanged. The stress decreased protein level of TH, as well as mRNA levels for NET and VMAT2 in the left stellate ganglia, but expressed no effect on protein levels of these two transporters. These results demonstrate asymmetry in noradrenergic genes in the right and left stellate ganglia during stress and provide molecular evidence to help explain the difference in response to the stress.

Peripheral oxytocin treatment affects the rat adreno-medullary catecholamine content modulating expression of vesicular monoamine transporter 2.

The neuropeptide oxytocin has been shown to influence on neuroendocrine function. The aim of the present study was to investigate the effect of peripheral oxytocin treatment on the synthesis, uptake and content of adreno-medullary catecholamine. For this purpose oxytocin (3.6µg/100g body weight, s.c) was administrated to male rats once a day over 14 days. In order to assess the effect of peripheral oxytocin treatment on adreno-medullary catecholamine we measured epinephrine and norepinephrine content and gene expression of tyrosine hydroxylase (TH), norepinephrine transporter (NET) and vesicular monoamine transporter 2 (VMAT2) in the adrenal medulla. Our results show a significant increase of epinephrine (1.7-fold, p<0.05) and norepinephrine (1.5-fold, p<0.05) content in oxytocin treated animals compared to saline treated ones. Oxytocin treatment had no effect either on mRNA or protein level of TH and NET. Under oxytocin treatment the increase in VMAT2 mRNA level was not statistically significant, but it caused a significant increase in protein level of VMAT2 (3.7-fold, p<0.001). These findings indicate that oxytocin treatment increases catecholamine content in the rat adrenal medulla modulating VMAT2 expression.

Molecular basis of chronic stress-induced hippocampal lateral asymmetry in rats and impact on learning and memory.

Neurochemical lateralization has been demonstrated in the rat brain suggesting that such lateralization might contribute to behavior. Thus, the aim of the present study was to examine neurochemical asymmetry in the hippocampus, molecular basis of neurochemical lateralization and its impact on spatial learning and memory. Changes in noradrenaline content, tyrosine hydroxylase (TH) were studied in the right and left hippocampus of naive control and chronically isolated rats, by applying TaqMan RT-PCR and Western blot analysis. Hippocampal-based spatial learning and memory were evaluated using the Barnes maze. In control rats an asymmetrical right-left distribution of noradrenaline content and gene expression of catecholamine synthesizing enzyme was found. Chronic psychosocial stress further emphasized asymmetry. Isolation stress reduced noradrenaline content only in the right hippocampus. No changes were observed in gene expression and protein levels of TH in the right hippocampus, whereas expression of catecholamine synthesizing enzyme was elevated in the left hippocampus. Reduced noradrenaline content in the right hippocampus did not cause impairment in spatial learning and memory. Our findings suggest that chronic psychosocial stress reduces noradrenaline stores in the right hippocampus which may be caused by molecular asymmetry, but it does not affect spatial learning and memory.

Regulation of atrial catecholamine enzymes and adrenoceptor gene expression after chronic stress.

Chronic stress is a risk factor for the development of numerous psychopathological conditions in humans including depression. Changes in gene expression of tyrosine-hydroxylase (TH), dopamine-ß-hydroxylase (DBH) phenylethanolamine N-methyltransferase (PNMT), ß1-, ß2- and ß3-adrenoceptors in right and left rat atria upon chronic unpredictable mild stress (CMS) were investigated. CMS decreased TH and DBH gene expression levels both in right and left atria and increased PNMT mRNA in left atria. No changes in mRNA levels of ß1- and ß2-adrenoceptors were recorded, whereas ß3-adrenoreceptor mRNA level was significantly elevated in right atria of CMS rats. At the same time, CMS produced a significant increase of ß1- and ß2-adrenoreceptor mRNA levels in left atria, but did not affect ß3-adrenoceptor mRNA level.The results presented here suggest that stress-induced depression expressed differential effects on catecholamine biosynthetic enzymes and ß-adrenoceptors at molecular level in right and left atria of adult rat males. Elevated gene expression of PNMT in left atria of rats exposed to CMS can lead to altered physiological response and may play a role in the pathophysiology of cardiovascular function.

Flouxetine treatment acts selectively increasing myocardial beta1-adrenoceptor mRNA expression in stress-induced depression.

Changes in gene expression of beta1- and beta2-adrenoceptors (beta1 - and beta2-AR) in right and left atria and ventricles after fluoxetine treatment in stress-induced depression of adult rat males were studied. Elevated beta1-AR mRNA levels in the left atria and significantly higher levels of beta2-AR mRNA in the left atria and ventricles were observed in stress-induced depression in comparison with those of unstressed controls. Fluoxetine treatment led to increasing expression of beta1-AR mRNA in the right atria and left ventricles, while the level of beta2-AR mRNA remained unchanged. These findings suggest that fluoxetine therapy plays an important role in cardiac beta-adrenergic subsensitivity and gene regulation of beta-AR in animals with heightened sympathetic nervous activity.

Regulation of catecholamine-synthesising enzymes and beta-adrenoceptors gene expression in ventricles of stressed rats.

Stress exposure activates the sympathoneural system, resulting in catecholamine release. Chronic stress is associated with development of numerous disorders, including cardiovascular diseases. Here we investigated the expression of mRNAs for catecholamine biosynthetic enzymes tyrosine-hydroxylase, dopamine-beta-hydroxylase and phenylethanolamine N-methyl-transferase, and for beta(1)- and beta(2)-adrenoceptors in the right and left ventricles of rats exposed to chronic unpredictable mild stress. The tyrosine-hydroxylase and dopamine-beta-hydroxylase mRNA levels were not affected by stress, whereas the phenylethanolamine N-methyltransferase mRNA levels significantly increased in both right and left ventricles. No changes in beta(1)-adrenoceptor mRNA levels in either right or left ventricles were observed. At the same time, stress produced a significant increase of beta(2)-adrenoceptor mRNA levels in left ventricles. These results suggest that elevated expression of phenylethanolamine N-methyltransferase in both ventricules and beta(2)-adrenoceptor genes in left ventricles could provide a molecular mechanism that leads to altered physiological response, which is important for the organism coping with stress.

Effects of repeated maprotiline and fluoxetine treatment on gene expression of catecholamine synthesizing enzymes in adrenal medulla of unstressed and stressed rats.

1 Repeated maprotiline (a noradrenaline reuptake inhibitor) and fluoxetine (a serotonin reuptake inhibitor) treatment on gene expression of catecholamine biosynthetic enzymes were examined in adrenal medulla of unstressed control and chronic unpredictable mild stressed rats. 2 Maprotiline did not change gene expression of catecholamine biosynthetic enzymes in control and stressed rats. 3 Fluoxetine increased gene expression of tyrosine hydroxylase (TH) and dopamine-ß-hydroxylase (DBH), but did not phenylethanolamine N-methyltransferase in both unstressed and chronic unpredictable mild stressed animals. 4 In conclusion, we have demonstrated that repeated administration of fluoxetine enhanced gene transcription of TH and DBH and subsequently stimulates noradrenaline synthesis in adrenal medulla of control and stressed rats.

Effect of immobilization stress on gene expression of catecholamine biosynthetic enzymes in heart auricles of socially isolated rats

Chronic stress is associated with the development of cardiovascular diseases. The sympathoneural system plays an important role in the regulation of cardiac function both in health and disease. In the present study, the changes in gene expression of the catecholamine biosynthetic enzymes tyrosine hydroxylase (TH), dopamine-â-hydroxylase (DBH) and phenylethanolamine N-methyltransferase (PNMT) and protein levels in the right and left heart auricles of naive control and long-term (12 weeks) socially isolated rats were investigated by Taqman RT-PCR and Western blot analysis. The response of these animals to additional immobilization stress (2 h) was also examined. Long-term social isolation produced a decrease in TH mRNA level in left auricles (about 70 percent) compared to the corresponding control. Expression of the DBH gene was markedly decreased both in the right (about 62 percent) and left (about 81 percent) auricles compared to the corresponding control, group-maintained rats, whereas PNMT mRNA levels remained unchanged. Exposure of group-housed rats to acute immobilization for 2 h led to a significant increase of mRNA levels of TH (about 267 percent), DBH (about 37 percent) and PNMT (about 60 percent) only in the right auricles. Additional 2-h immobilization of individually housed rats did not affect gene expression of these enzymes in either the right or left auricle. Protein levels of TH, DBH and PNMT in left and right heart auricles were unchanged either in both individually housed and immobilized rats. The unchanged mRNA levels of the enzymes examined after short-term immobilization suggest that the catecholaminergic system of the heart auricles of animals previously exposed to chronic psychosocial stress was adapted to maintain appropriate cardiovascular homeostasis.

Effect of immobilization stress on gene expression of catecholamine biosynthetic enzymes in heart auricles of socially isolated rats.

Chronic stress is associated with the development of cardiovascular diseases. The sympathoneural system plays an important role in the regulation of cardiac function both in health and disease. In the present study, the changes in gene expression of the catecholamine biosynthetic enzymes tyrosine hydroxylase (TH), dopamine-beta-hydroxylase (DBH) and phenylethanolamine N-methyltransferase (PNMT) and protein levels in the right and left heart auricles of naive control and long-term (12 weeks) socially isolated rats were investigated by Taqman RT-PCR and Western blot analysis. The response of these animals to additional immobilization stress (2 h) was also examined. Long-term social isolation produced a decrease in TH mRNA level in left auricles (about 70%) compared to the corresponding control. Expression of the DBH gene was markedly decreased both in the right (about 62%) and left (about 81%) auricles compared to the corresponding control, group-maintained rats, whereas PNMT mRNA levels remained unchanged. Exposure of group-housed rats to acute immobilization for 2 h led to a significant increase of mRNA levels of TH (about 267%), DBH (about 37%) and PNMT (about 60%) only in the right auricles. Additional 2-h immobilization of individually housed rats did not affect gene expression of these enzymes in either the right or left auricle. Protein levels of TH, DBH and PNMT in left and right heart auricles were unchanged either in both individually housed and immobilized rats. The unchanged mRNA levels of the enzymes examined after short-term immobilization suggest that the catecholaminergic system of the heart auricles of animals previously exposed to chronic psychosocial stress was adapted to maintain appropriate cardiovascular homeostasis.

[The effect of Hatha yoga on poor posture in children and the psychophysiologic condition in adults]. / Delovanje Hata joge na lose drzanje tela kod dece i psihofizicku kondiciju kod odraslih.

Hatha Yoga's effects on the posture of 15 ten year-old children and also its effects on the psychophysical condition of 15 grown-ups was studied. As symptoms, during the first examination, 12 of the 15 children had head protrusion, 14 had shortened back extensors, all 15 had bent shoulders, relaxation of the frontal abdominal wall and shortened flexors of both the calf and thigh. The condition of all the children was remarkably better after six months of practice, some of the symptoms having completely disappeared (head protrusion, asymmetry of the shoulders, mamillas and hips, shortening of the pectoralis and back extensors), 9 children still had slight to medium relaxation of the frontal abdominal wall, 8 children still had bent shoulders, and 1 child still had shortened calf and thigh extensors. The adults were in a weak or very weak psychophysical condition, they tired easily, they complained of sleep disturbances, fluctuation of emotional state and irritability. After 3 months of practice, the vital capacity of 8 of the adults tested (53.3%) had increased by 435 ml. The time duration of apnoea had lengthened for all of the practicing adults, but with a truly large variation among them (a median of 14%). The deep waist-bend length of all the practicing adults had lengthened by an average of 9.5 cm, and the average length increase for the 3-minute running test was 42 m. All those who practiced, had experienced an alleviation of psychic difficulties.