Natasa Hlavacova

Chronic treatment with the mineralocorticoid hormone aldosterone results in increased anxiety-like behavior

Aldosterone is the last component of the renin-angiotensin-aldosterone system inducing its peripheral effects via mineralocorticoid receptors (MR). Brain MR bind preferentially glucocorticoids. So far, the role of MR in behavioral functions has been investigated almost exclusively in relation to glucocorticoids. Recently, aldosterone itself has been linked to affective disorders. The aim of this study was to test the hypothesis that chronic elevation of circulating levels of aldosterone leads to increased anxiety. We have investigated the effects of chronic aldosterone treatment on (1) anxiety-like behavior, and (2) basal and stress-induced levels of selected hormones. Forty male Wistar rats were subcutaneously implanted with osmotic minipumps and treated with aldosterone (2 microg/100 g/day) or vehicle for two weeks. Aldosterone concentrations in plasma showed a mild (approximately four-fold) increase at the end of two-week aldosterone treatment. This mild hyperaldosteronism resulted in a significant enhancement of anxiety as demonstrated by alterations in all indicators of anxiety-like behavior measured in the open field and elevated plus-maze tests, without significant changes in measures of general locomotor activity. Aldosterone treatment affected not only the spatiotemporal measures of anxiety, but also the ethological parameters related to exploration and risk assessment. Chronic treatment with aldosterone was associated with increased water intake and decreased plasma renin activity, but failed to modify basal or stress-induced activity of the hypothalamic-pituitary-adrenocortical axis. The results provide evidence on anxiogenic action of prolonged increase in circulating aldosterone concentrations. Thus, aldosterone may represent an important target for future antidepressant and anxiolytic drug development.

Eplerenone, a selective mineralocorticoid receptor blocker, exerts anxiolytic effects accompanied by changes in stress hormone release

The aim of this study was to investigate the impact of chronic treatment with eplerenone, a mineralocorticoid receptor antagonist and clinically used antihypertensive drug, on animal correlates of mood disorders, namely anxiety-like behaviour, stress hormones release and brain plasticity. Male rats (n = 40) were injected subcutaneously twice daily with eplerenone (50 mg/kg body weight) or vehicle for 11 days. Open-field and elevated plus-maze tests were used as both anxiety-related paradigms and stress stimuli to evaluate hormone responses. Eplerenone-treated rats showed reduced anxiety-like behaviour manifested by both conventional and ethological parameters related to exploration and risk assessment behaviour in the elevated plus-maze test and partially in the open-field test. Eplerenone treatment resulted in an elevation of plasma aldosterone and oxytocin levels. Chronic treatment with eplerenone prevented the stress-induced rise in plasma corticosterone levels and vasopressin concentrations in the posterior pituitary. Eplerenone treatment failed to induce substantial changes in hippocampal brain derived neurotrophic factor protein concentrations. In conclusions, chronic treatment with eplerenone (1) exerts anxiolytic effects and (2) influences corticosterone, oxytocin and vasopressin concentrations in a manner consistent with the anxiolytic outcome.

Endocrine Factors in Stress and Psychiatric Disorders

Glucocorticoids and other steroids produced in the adrenal cortex are altered in chronic stress situations associated with enhanced anxiety. A useful tool to evaluate changes in adrenal steroids during stress and anxiety under both laboratory and real‐life stress situations is determination of steroids in saliva. The main advantages of this technique are its noninvasiveness and its measurement of biologically active free hormone levels. Salivary cortisol is a valuable indicator of the activity of the hypothalamic‐pituitary‐adrenocortical axis, which is known to be altered in psychiatric disorders. Measurements of salivary cortisol helped to reveal changes that would otherwise remained undetected, such as increase in cortisol release in spontaneously occurring panic attacks. By selecting only the subjects with high and low trait anxiety, we have brought evidence confirmed by others that high trait anxiety may be associated with an inability to respond with adequate cortisol release during stress. Papers on the relationship between salivary dehydroepiandrosterone and trait anxiety or anxiety disorders are rare, and this stress hormone deserves more attention. Almost nothing is known on aldosterone and anxiety. We have modified the methodology of aldosterone radioimmunoassay by concentrating the saliva and validated it biologically by demonstrating daily variation and gender differences. We have provided the first data on the relationship between aldosterone and trait anxiety. Obtained results show a significant negative correlation between morning salivary aldosterone concentrations and trait anxiety scores in women (luteal phase), but not in men. A more proper elucidation of the association between aldosterone and anxiety seems to be an important target of further research.

Subchronic treatment with aldosterone induces depression-like behaviours and gene expression changes relevant to major depressive disorder.

The potential role of aldosterone in the pathophysiology of depression is unclear. The aim of this study was to test the hypothesis that prolonged elevation of circulating aldosterone induces depression-like behaviour accompanied by disease-relevant changes in gene expression in the hippocampus. Subchronic (2-wk) treatment with aldosterone (2 μg/100 g body weight per day) or vehicle via subcutaneous osmotic minipumps was used to induce hyperaldosteronism in male rats. All rats (n = 20/treatment group) underwent a modified sucrose preference test. Half of the animals from each treatment group were exposed to the forced swim test (FST), which served both as a tool to assess depression-like behaviour and as a stress stimulus. Affymetrix microarray analysis was used to screen the entire rat genome for gene expression changes in the hippocampus. Aldosterone treatment induced an anhedonic state manifested by decreased sucrose preference. In the FST, depressogenic action of aldosterone was manifested by decreased latency to immobility and increased time spent immobile. Aldosterone treatment resulted in transcriptional changes of genes in the hippocampus involved in inflammation, glutamatergic activity, and synaptic and neuritic remodelling. Furthermore, aldosterone-regulated genes substantially overlapped with genes affected by stress in the FST. This study demonstrates the existence of a causal relationship between the hyperaldosteronism and depressive behaviour. In addition, aldosterone treatment induced changes in gene expression that may be relevant to the aetiology of major depressive disorder. Subchronic treatment with aldosterone represents a new animal model of depression, which may contribute to the development of novel targets for the treatment of depression.

Oxytocin levels in the posterior pituitary and in the heart are modified by voluntary wheel running

We hypothesized that voluntary wheel running results in increased secretion of oxytocin, a peptide involved in the stress response. An additional hypothesis was that prolonged exercise affects oxytocin levels in the heart, which is in line with the potential role of oxytocin in cardiovascular functions. Voluntary wheel running lasted 3 weeks and daily running distances increased progressively reaching maximum levels about 8 km (Sprague-Dawley rats) and 4 km (Lewis strain). The exercise resulted in significant reduction of epididymal fat, slight increase in glucose transporter GLUT4 mRNA levels and significant enhancement of plasma density. Voluntary exercise failed to influence plasma oxytocin levels either in Lewis or Sprague-Dawley rats, but it resulted in a significant decrease of oxytocin concentrations in the posterior pituitary. Plasma oxytocin concentrations were not modified even if the measurements were made in the dark phase of the day. In voluntary wheel running Sprague-Dawley rats, the content of oxytocin in the right heart atrium was lower than in controls. Thus, the present findings demonstrate that prolonged voluntary wheel running results in a decrease in pituitary oxytocin content without evident changes in hormone concentrations in peripheral blood. However, prolonged exercise used has a significant impact on oxytocin levels in the heart.

Neuroendocrine Activation during Combined Mental and Physical Stress in Women Depends on Trait Anxiety and the Phase of the Menstrual Cycle

The aim of this study was to investigate the influence of trait anxiety and menstrual cycle phase on neuroendocrine activation during combined mental and physical stress procedure in 40 healthy female subjects. Women at the upper (anxious) and lower (nonanxious) limits of the normal range of a trait anxiety scale were exposed to the stress procedure consisting of a mental component (Stroop test) and handgrip exercise. Salivary cortisol levels, cardiovascular parameters, and cognitive performance in the Stroop test were evaluated. Stress‐induced cortisol levels and the rise in systolic blood pressure were affected by both trait anxiety and menstrual cycle phase. The stress model used induced a significant cortisol elevation only in anxious women in the follicular phase. This group of women also exhibited greater increases in systolic blood pressure in response to handgrip exercise as compared to anxious ones in the luteal phase and to nonanxious women in either phase. In nonanxious women, stress‐induced cortisol levels positively correlated with cognitive performance. In contrast, a negative correlation trend was observed in anxious subjects. Thus, in subjects with low but not high trait anxiety, enhanced cortisol concentrations seem to be associated with better cognitive performance. The results suggest that women with high trait anxiety exhibit greater cardiovascular and hormonal sensitivity to stress stimuli during the follicular phase.

Attenuated neuroendocrine response to hypoglycemic stress in patients with panic disorder.

Background/Aims: There is a lack of information on the effects of metabolic stress exposure on hormone release in patients with panic disorder. The aim of this study was to test the hypothesis that neuroendocrine activation during hypoglycemic stress is altered in panic disorder patients compared to healthy subjects. Methods: Hormone responses to an intravenous bolus of insulin (0.1 IU/kg) were evaluated in both fully remitted, medication-free panic disorder patients and healthy controls (n = 9/group). Blood samples for determination of cortisol, growth hormone, prolactin, adrenaline and noradrenaline concentrations were obtained at rest and over a 90-min period after insulin injection. Results: In patients with panic disorder, basal prestress hormone levels were comparable to those in healthy subjects with the exception of plasma adrenaline, which was higher in panic disorder patients compared to controls. The degree of hypoglycemia induced by insulin administration was similar in patients and healthy subjects. Hypoglycemia-induced increases in growth hormone, prolactin and cortisol concentrations were significantly attenuated in panic disorder patients compared to healthy individuals. No differences between patients and controls were observed in adrenalin release induced by hypoglycemia. Conclusions: The present data demonstrate that somatotropic, lactotropic and corticotropic activation during hypoglycemic stress is blunted in patients with panic disorder. It is suggested that in contrast to the effects of relatively mild stress conditions used in other studies published in the literature, intensive stressors inducing a broad spectrum of hormonal changes fail to provoke an adequate neuroendocrine response in patients with panic disorder.

Aldosterone Signals the Onset of Depressive Behaviour in a Female Rat Model of Depression along with SSRI Treatment Resistance.

Depression is a serious condition that occurs more frequently in women and is often associated with treatment resistance. The main hypotheses of this study are that (a) aldosterone is an early marker of depression onset and (b) a tryptophan (TRP) depletion model of depression previously validated in male rats is treatment resistant in females. To explore possible underlying mechanisms, we have focused on factors shown to be altered in patients with treatment-resistant depression. Female Sprague-Dawley rats were treated with a control or low-TRP-containing diet for various time periods up to 21 days. The results show that aldosterone secretion increased after 4 days of TRP depletion and prior to corticosterone. Optimal effects of TRP depletion occurred at 14 days. In addition to neurochemical and behavioural changes observed previously in males, TRP depletion in females was associated with a significant decline in serum magnesium concentrations, increased serum interleukin-6, enhanced gene expression of orexin A in the frontal cortex and induced a rise in N-methyl-D-aspartate (NMDA) receptor Bmax in the amygdala. Depression-like behaviour, NMDA receptor upregulation, enhancement of the kynurenine-to-kynurenic acid ratio and magnesium were resistant to paroxetine treatment (10 mg/kg/day in drinking water for 14 days). In conclusion, aldosterone may represent an important early marker for the onset of depression-like behaviour. With respect to treatment resistance, the underlying mechanisms may involve pro-inflammatory cytokines, the kynurenine pathway, magnesium, glutamate neurotransmission and the orexin pathway. This model of treatment-resistant depression may be useful for the future development of new compounds with novel antidepressant properties.

Increased anxiety induced by listening to unpleasant music during stress exposure is associated with reduced blood pressure and ACTH responses in healthy men.

The relationship between anxiety and the neuroendocrine response to stress stimuli is still not fully understood. The aim of this study was to evaluate the contribution of an acute increase in state anxiety to neuroendocrine activation under stress conditions. To do so, it was necessary to find a stress condition of the same character and intensity with and without a rise in state anxiety. We decided to examine the effects of listening to music on anxiety and to apply a new methodological approach. A group of 14 healthy volunteers participated in a counterbalanced crossover design study. The stress procedure consisted of mental (Stroop test, mental arithmetic) and physical (handgrip exercise) tasks combined with listening to music played forward (pleasant) or backwards (unpleasant). The results confirmed our hypothesis, namely the condition with listening to unpleasant music was anxiogenic, while the other was not. In case of increased state anxiety, the rise in ACTH concentrations in response to mental challenge and the increase in systolic blood pressure induced by handgrip exercise was reduced compared to the situation with unchanged anxiety. Concentrations of testosterone, oxytocin, vasopressin and aldosterone were slightly increased in response to the stress paradigm accompanied with increased anxiety. In conclusion, the present data demonstrate that an acute increase in state anxiety contributes to neuroendocrine activation under stress conditions. Moreover, the results show that listening to music may both positively and negatively influence the perception of stress and the level of anxiety, which might have functional consequences.

The Evidence for Altered BDNF Expression in the Brain of Rats Reared or Housed in Social Isolation: A Systematic Review

There is evidence that development and maintenance of neural connections are disrupted in major mental disorders, which indicates that neurotrophic factors could play a critical role in their pathogenesis. Stress is a well-established risk factor for psychopathology and recent research suggests that disrupted signaling via brain-derived neurotrophic factor (BDNF) may be involved in mediating the negative effects of stress on the brain. Social isolation of rats elicits chronic stress and is widely used as an animal model of mental disorders such as schizophrenia and depression. We carried out a systematic search of published studies to review current evidence for an altered expression of BDNF in the brain of rats reared or housed in social isolation. Across all age groups (post-weaning, adolescent, adult), majority of the identified studies (16/21) reported a decreased expression of BDNF in the hippocampus. There are far less published data on BDNF expression in other brain regions. Data are also scarce to assess the behavioral changes as a function of BDNF expression, but the downregulation of BDNF seems to be associated with increased anxiety-like symptoms. The reviewed data generally support the putative involvement of BDNF in the pathogenesis of stress-related mental illness. However, the mechanisms linking chronic social isolation, BDNF expression and the elicited behavioral alterations are currently unknown.