Biomedical subjects

R Kvetnanský

Publications and source records attributed to R Kvetnanský.

At least 19 recordsLinked to original sources

PubMed2006

The effects of stress on muscarinic receptors. Heterologous receptor regulation: yes or no?

1 Stress is usually comprehended as an event affecting mainly the catecholaminergic system, the hypothalamo-pituitary-adrenocortical (HPA) axis and the receptor systems connected to these neurotransmitters/hormones. Other neurotransmitter/hormone systems can be affected too. Here we review the available data on the effects of different stressful stimuli (physical, chemical, psychological/social, cardiovascular, affecting multiple system) on muscarinic receptors (MR). 2 The data suppose the existence of specific mechanisms that regulate the signalization through MR during different type of stress. 3 Physical stressors (cold vs. heat) reveal opposite type of changes on peripheral-tissue MRs. Chemical stressors (oxidative stress) are tightly connected with MR and it is especially interesting that the sensitivity of MR to oxidative stress is subtype-specific. It is also suggested that heterologous regulation can occur with psychological/social stressors on the organism. Cardiovascular system-disturbing stressors cause imbalance between autonomic receptors or down-regulate MR in the peripheral tissue. Immobilization caused opposite effects on MR in the central nervous system and periphery, where the changes are supposed to be due to heterologous regulation between receptor systems. 4 In conclusion, some data indicate that in specific conditions MR are regulated as a consequence of other changes rather than as a primary effect of stress. On the contrary, in some situations, MR are the first targets to respond to the stress. 5 These findings on stress-induced activity of the cholinergic system and changes in muscarinic receptors support the view that stress is a specific response of the organism.

Animals↗

PubMed2006

Developmental changes of some G-protein coupled receptors affected by c-fos knock-out.

The role of c-fos gene disruption on binding characteristics of selected G protein-coupled receptors has been investigated. The following receptors were studied: muscarinic receptors (MR), alpha1-adrenoceptors (AAR), beta-adrenoceptors (BAR), D1-like dopamine receptors (D1R), D2-like dopamine receptors (D2R). We have tested the hypothesis that c-fos gene disruption would not influence the receptor density in central nervous system (CNS: brain cortex and cerebellum) and in the periphery (lung, heart). We found that both in the periphery and in the CNS there were important changes in receptor density. Both MR and BAR were increased in the lung and heart. The effects of c-fos gene disruption in CNS were more selective. In general, the receptors that activate Gq-phospholipase C-proteinkinase C pathway (AAR, MR) were affected, while the others (that activate/or inhibit adenylyl cyclase: BAR, D1R, D2R) were not. These results show that disruption of c-fos gene could significantly change the expression of G protein-coupled receptors. Moreover, these changes could be comprehended as one of the adaptive mechanisms that help the organism to cope with c-fos gene disruption.

Animals↗

PubMed2004

Mapping of genetic determinants of the sympathoneural response to stress.

Activation of the sympathoadrenal system (SAS, comprising the sympathetic nervous system and the adrenal medulla) in response to stressful stimuli is an important defense mechanism as well as a contributor to several cardiovascular diseases. There is variability in the SAS response to stress, although the extent to which this is genetically regulated is unclear. Some rodent models, including the hereditary hypertriglyceridemic (hHTg) rat, are hyperresponsive to stress. We investigated whether quantitative trait loci (QTLs) that affect sympathoadrenal response to stress could be identified. Second filial generation rats (n = 189) derived from a cross of the hHTg rat and the Brown Norway rat had plasma norepinephrine (NE) and epinephrine (Epi) levels, indices of activation of the sympathoneural and adrenal medulla components, respectively, measured in the resting state and in response to an immobilization stress. Responses were assessed early (20 min) and late (120 min) after the application of the stress. A genome scan was conducted using 153 microsatellite markers. Two QTLs (maximum peak LOD scores of 4.17 and 3.52, respectively) influencing both the early and late plasma NE response to stress were found on chromosome 10. Together, the QTLs accounted for approximately 20% of the total variation in both the early and late NE responses in the F(2) rats. Interestingly, the QTLs had no effect on plasma Epi response to stress. These findings provide evidence for a genetic determination of the response of a specific component of the SAS response to stress. Genetically determined variation in sympathetic nervous system response to stress may contribute to cardiovascular diseases.

Animals↗

PubMed2004

Effect of immobilization on in vitro thyrotropin-releasing hormone release from brain septum in wild-type and corticotropin-releasing hormone knock-out mice.

There is considerable evidence linking alcohol consumption, sedation, and thyrotropin-releasing hormone (TRH) in the brain septum. We have shown that ethanol in clinically relevant concentrations can in vitro induce TRH release from the septum by a mechanism involving neuronal swelling. Corticotropin-releasing hormone-deficient (CRH-KO) mice serve as an interesting model to help us understand the role of CRH in the regulation of different neuroendocrine systems. The aim of this study was to compare TRH release activity in the brain septum at basal and stress conditions in CRH-KO mice and their wild-type (WT) littermates. Experimental mice were decapitated immediately or 3 h after single (2 h) or repeated (seven times for 2 h daily) immobilization stress. The brain septum was immediately cut out and incubated to measure basal-, ethanol-, and hyposmosis-stimulated TRH release in vitro. Ethanol in isosmotic medium or hyposmotic medium stimulated TRH release from mice septal explants from WT and CRH-KO mice. The response was disturbed immediately after immobilization and recovered 3 h later. Our results show that immobilization stress transiently affects the TRH system in brain septum. Inborn absence of CRH does not affect septal TRH and its response to ethanol before and 3 h after immobilization.

Animals↗

PubMed2004

Effect of space flight and head-down bedrest on neuroendocrine response to metabolic stress in physically trained subjects.

The aim of this study was to evaluate the association of plasma epinephrine (EPI) and norepinephrine (NE) responses to insulin induced hypoglycemia (ITT) 3 weeks before the space flight (SF), on the 5th day of SF, on the 2nd and 16th days after the landing in the first Slovak astronaut, and before and on the 5th day of prolonged subsequent head-down (-6 degrees) bed rest (BR) in 15 military aircraft pilots. Blood samples during the test were collected via cannula inserted into cubital vein, centrifuged in the special appliance Plasma-03, frozen in Kryogem-03, and at the end of the 8-day space flight transferred to Earth in special container for hormonal analysis. Insulin hypoglycemia was induced by i.v. administration of 0.1 IU/kg BW insulin (Actrapid HM) in bolus. Insulin administration led to a comparable hypoglycemia in pre-flight, in-flight conditions and before and after bed rest. ITT led to a pronounced increase in EPI levels and moderate increase in NE in pre-flight studies. However, an evidently reduced EPI response was found after insulin administration during SF and during BR. Thus, during the real microgravity in SF and simulated microgravity in BR, insulin-induced hypoglycemia activates the adrenomedullary system to less extent than at conditions of the Earth gravitation. Post-flight changes in EPI and NE levels did not significantly differ from those of pre-flight since SF was relatively short (8 days) and the readaptation to Earth gravitation was fast. It seems, that an increased blood flow in brain might be responsible for the reduced EPI response to insulin. Responses to ITT in physically fit subjects indicate the stimulus specificity of deconditioning effect of 5 days bed rest on stress response. Thus, the data indicate that catecholamine responses to ITT are reduced after exposure to real as well as simulated microgravity.

Adult↗

PubMed2003

Effect of head-down bed rest on the neuroendocrine response to orthostatic stress in physically fit men.

The role of neuroendocrine responsiveness in the development of orthostatic intolerance after bed rest was studied in physically fit subjects. Head-down bed-rest (HDBR, -6 degrees, 4 days) was performed in 15 men after 6 weeks of aerobic training. The standing test was performed before, after training and on day 4 of the HDBR. Orthostatic intolerance was observed in one subject before and after training. The blood pressure response after training was enhanced (mean BP increments 18+/-2 vs. 13+/- 2 mm Hg, p<0.05, means +/- S.E.M.), although noradrenaline response was diminished (1.38+/-0.18 vs. 2.76+/-0.25 mol.l(-1), p<0.01). Orthostatic intolerance after HDBR was observed in 10 subjects, the BP response was blunted, and noradrenaline as well as plasma renin activity (PRA) responses were augmented (NA 3.10+/-0.33 mol.l(-1), p<0.001; PRA 2.98+/-1.12 vs. 0.85+/-0.15 ng.ml(-1), p<0.05). Plasma noradrenaline, adrenaline and aldosterone responses in orthostatic intolerant subjects were similar to the tolerant group. We conclude that six weeks of training attenuated the sympathetic response to standing and had no effect on the orthostatic tolerance. In orthostatic intolerance the BP response induced by subsequent HDBR was absent despite an enhanced sympathetic response.

Adult↗

PubMed2003

[Use of the in vivo microdialysis technique in basic and clinical research].

Microdialysis is in vivo technique that permits monitoring of local concentrations of metabolites and drugs at specific sites in the body that makes it an attractive tool for the basic and clinical research. Microdialysis entered the experimental studies of the brain, and later also of the peripheral tissues such as adipose and muscle tissues, kidney, lung, eye, skin, and blood. There are several critical factors in the experimental implementations of microdialysis: the probe, the perfusion solution, tissue integrity, and the sensitivity of microdialysate analysis method. When the experimental conditions are optimised to give valid results, microdialysis can provide numerous data from the relatively small number of individual subjects. It can bring about detailed clinical information reflecting free metabolite and drug concentrations in studied tissues and/or in plasma. With the progress of analytical methods, applications and importance of the microdialysis technique in clinical and pharmacokinetic research and diagnostics will increase.

Humans↗

PubMed2003

[Use of knock-out mice in studies of stress reactions].

Modern human population intimately recognizes the stress reaction of the organism, which has developed as a response to variety physical and/or psychical stressors. This reaction represents not only a complex of adaptive mechanisms enabling the successful overcoming of the conflicting situation, but in its pathological form it can significantly contribute to the development of serious diseases, from carcinogenic, cardiovascular, gastrointestinal, up to the psychical. New, developing scientific disciplines permit to widen knowledge of the basis of this undesirable side of the stress. Transgenic animals, which either has integrated the gene of interest, or studied gene is not functioning (i.e. knockout animals), permit at least partially to study the physiological importance of the product of this gene in the organism. In our review we described in detail three lines of knockout mice--with the knockout gene for the dopamine-beta-hydroxylase, corticoliberin and c-fos--that significantly contribute to the regulation of the stress response.

Animals↗

PubMed2002

[Catecholamine synthesis and expression of enzymes of the catecholamine pathway in the rat heart at rest and during stress].

Catecholamines participate in a variety of cell regulations and physiological mechanisms, but also in the development of neurological, psychiatrical, endocrine and cardiovascular diseases. Diseases of the cardiovascular system, such as hypertension, myocardial infarction, ischaemia, etc. represent a serious medical problem and a frequent cause of the human death. It is alarming that development of these diseases depends preferentially on the wrong life-style and affect lower age groups. One of the factors that participate on unwilling increase of the incidence of cardiovascular diseases is the stress. Under the stress, organism mobilizes its energetical sources, increases cardiac performance and activates other processes, which enable to handle this load. Catecholamines belong to the important mediators of the stress response. On the one side they are required to buffer the stress situation, but the pathological changes in the regulation of their synthesis, secretion and action significantly participate on the development of several diseases (e.g. also of the cardiovascular origin). The aim of this review is to show the role of catecholamines in the heart not only under the physiological, but also in pathophysiological conditions. Stress as a modern civilization factor participates on the development of several diseases. Understanding of the mechanism of development of these diseases is the first step to the development of an appropriate therapy. Changes in the catecholamine levels during stress, as well as enzymes, which participate on the synthesis of catecholamines, are undoubtedly the important part of this mechanism.

Animals↗

PubMed2001

Identification of tyrosine hydroxylase gene expression in rat spleen.

This study was aimed to identify tyrosine hydroxylase (TH) gene expression in the rat spleen under basal and stress conditions. Using the reverse transcription polymerase chain reaction we did not detect TH mRNA in rat spleen either in control, or immobilized animals. Semi-nested PCR revealed a clear signal, demonstrating that TH mRNA is formed in the spleen, although in low abundance. We also detected both, TH immunoreactive protein and TH activity in the rat spleen that were in higher abundance than expected from the mRNA levels. This study identifies, for the first time, TH gene expression in rat spleen. Since TH protein and activity are present in the spleen in much higher abundance compared to corresponding mRNA, the majority of TH protein is most probably supplied by the sympathetic innervation of spleen.

Animals↗

PubMed2001

Effect of novel stressors on gene expression of tyrosine hydroxylase and monoamine transporters in brainstem noradrenergic neurons of long-term repeatedly immobilized rats.

Responses of central noradrenergic (NE) neurons to stressors like immobilization (IMO), cold exposure, insulin-induced hypoglycemia, and cellular glucoprivation caused by 2-deoxy-D-glucose (2-DG) were investigated in intact and long-term repeatedly immobilized (LTR, 2 h daily IMO for 41 days) rats. Expression of tyrosine hydroxylase (TH), norepinephrine transporter (NET) and vesicular monoamine transporter (VMAT2) genes were determined by using in situ hybridization histochemistry in brainstem A1, A2, A5 and locus coeruleus (LC) neurons. TH mRNA levels were increased by single IMO or 2-DG administration in all areas studied. Cold was effective only in LC and A2 neurons while insulin had no effect. LTR immobilization elevated TH mRNA levels in all investigated cell groups. These elevations were equally high to those elicited by a single IMO in each noradrenergic group, except the LC where LTR IMO was less effective than the single IMO. The levels of NET and VMAT2 mRNAs were elevated only in the A1 and A2 cell groups of LTR IMO rats. A newly applied IMO in LTR rats did not alter TH, NET, and VMAT2 mRNA levels in any NE cell group investigated. Novel stressors like cold and 2-DG exaggerated the increased TH mRNA levels only in the LC of LTR IMO rats, unlike in the other NE cell groups. The present data indicate that repeated exposure of rats to homotypic stressor induces an adaptation of NE neurons, whereas single exposure of such animals to heterotypic novel stressor produces an exaggerated response of the system at the level of TH (in LC) and NET (in A1, A2) gene expression.

Animals↗

PubMed2001

Stress-triggered activation of gene expression in catecholaminergic systems: dynamics of transcriptional events.

Stress triggers important adaptive responses that enable an organism to cope with a changing environment. However, when prolonged or repeated, stress can be extremely harmful. The release of catecholamines is a key initial event in responses to stressors and is followed by an increase in the expression of genes that encode catecholamine-synthesizing enzymes. This process is mediated by transcriptional mechanisms in the adrenal medulla and the locus coeruleus. The persistence of transcriptional activation depends on the duration and repetition of the stress. Recent work has begun to identify the various transcription factors that are associated with brief or intermediate duration of a single or repeated stress. These studies suggest that dynamic interplay is involved in converting the transient increases in the rate of transcription into prolonged (potentially adaptive or maladaptive) changes in gene expression.

Adrenal Medulla↗

PubMed2001

Altered function of peripheral organ systems in rats exposed to chronic mild stress model of depression.

1. In depression, psychiatric symptoms are frequently associated with impaired cardiovascular function and perhaps also increased risk for cancer diseases. Pathophysiological basis of this comorbidity is not clearly understood. Molecular events involved, particularly factors modified by chronic stress exposure, may only be evaluated in animal models of depression. 2. Present experiments were aimed to study parameters related to cardiovascular system (tyrosine hydroxylase (TH) gene expression in adrenal glands) and carcinogenesis (retinoic acid receptors in the liver) in the chronic mild stress model of depression. 3. Chronic mild stress induced a rise in adrenal TH gene expression in both male and female rats. Gender dependent changes were found in retinoic acid receptor binding with stress-induced activation in females but not males. Ovariectomized animals exhibited higher retinoic acid receptor binding. slightly elevated TH mRNA levels and failed to respond to chronic mild stress exposure with further increase in TH mRNA levels. Similarly, chronic mild stress induced an anhedonic state manifested by decreased sucrose preference in control but not ovariectomized rats. 4. Presented data document that central neurochemical and behavioral changes in animals exposed to chronic mild stress model of depression are associated with changes in adrenal TH gene expression and with gender dependent changes in retinoic acid receptor status in the liver. Such alterations may participate in the development of pathological changes and could participate on increased risk for cardiovascular and oncologic comorbidity in depressive patients.

Adrenal Medulla↗

PubMed2001

Existence of cardiac PNMT mRNA in adult rats: elevation by stress in a glucocorticoid-dependent manner.

Phenylethanolamine N-methyltransferase (PNMT) is the enzyme that synthesizes epinephrine from norepinephrine. The aim of this study was to determine potential PNMT gene expression in the cardiac atria and ventricles of adult rats and to examine whether the gene expression of this enzyme is affected by immobilization stress. PNMT mRNA levels were detected in all four parts of the heart, with the highest level in the left atrium. Both Southern blot and sequencing verified the specificity of PNMT detected by RT-PCR. Single immobilization for 2 h increased gene expression of PNMT in both atria and ventricles. In atria, this effect was clearly modulated by glucocorticoids, because either adrenalectomy or hypophysectomy prevented the increase in PNMT mRNA levels in response to immobilization stimulus. This study establishes, for the first time, that PNMT gene expression occurs in cardiac atria and also, to a small extent, in ventricles of adult rats. Immobilization stress increases gene expression in atria and ventricles. This increase requires an intact hypothalamus-pituitary-adrenocortical axis, indicating the involvement of glucocorticoids.

Adrenalectomy↗

PubMed2001

Repeated immobilization stress reduces rat vertebral bone growth and osteocalcin.

We previously showed that psychological stressors alter plasma levels of osteocalcin (pOC), a bone-specific mineral binding protein, in ways that differ with the type of stressor. To determine effects of chronic stress, we examined vertebrae, pOC, and corticosterone levels from conscious rats subjected to foot-restraint immobilization (Immo) daily for 1-42 times. After 40-42 Immo, basal pOC was decreased by 25% compared with unstressed rats, and the subsequent rise in pOC during Immo was blunted. Corticosterone was elevated 10-fold during Immo. Immo for seven times did not change vertebral OC concentration, but caused a slight decrease in calcium and phosphorous concentrations in younger rats. Rats Immo for 42 times exhibited reduced body weight, vertebral weight, and vertebral OC concentration but no significant differences in vertebral mineral concentrations. Body fat content was visibly decreased. We do not know the source of or the stimulus for the initial rise in pOC. We conclude that both decreased growth and bone OC concentration are due to repeatedly elevated stress hormones.

Animals↗

PubMed2000

Identification of type 1 IP(3) receptors in the rat kidney and their modulation by immobilization stress.

Inositol 1,4,5-trisphosphate receptor (IP(3)-receptor) is a calcium channel, transporting calcium from intracellular stores to the cytoplasm. In kidney, IP(3)-receptors are involved in the signal transduction of various hormones. In our work we studied the effect of immobilization stress on the IP(3)-receptor's protein content in renal cortex and the medulla of normotensive and hypertensive rats. We detected both mRNA and type 1 IP(3)-receptor protein in medulla, but not in renal cortex. We found that this receptor was approximately twice as abundant in normotensive as in genetically hypertensive rat kidney. Immobilization stress decreased the amount of type 1 IP(3)-receptor in the renal medulla of normotensive rats approximately five times, while no effect due to single and/or repeated stress was observed in the renal medulla of spontaneously hypertensive rats. The results indicate that expression of type 1 IP(3)-receptor in renal medulla is modulated by hypertension and immobilization stress.

Alternative Splicing↗

PubMed2000

Nuclear all-trans retinoic acid receptor status in rat liver: a comparison of effects of three different stressors--immobilization, laparotomy, and 2-deoxy-D-glucose.

Retinoic acids and their cognate nuclear receptors exert a substantial regulatory role in cell growth and development as well as in the neuroendocrine system. These effects are primarily mediated by all-trans retinoic acid receptors (RARs), members of the steroid/thyroid hormone receptor superfamily of ligand inducible transcription factors. The present study was undertaken in order to compare the effects of immobilization stress (IMO), laparotomy, and 2-deoxy-D-glucose (2DG)-induced intracellular glucopenia on both nuclear RAR affinity and concentration in the rat liver. IMO when compared to non-stressed group of rats, significantly reduced the RAR maximal binding capacity (Bmax) in liver, with the equilibrium association constant (Ka) remaining unchanged. No significant changes of the RAR Bmax and the Ka, were observed in liver of rats that underwent laparotomy. In contrast, a single dose of 2DG (500 mg/kg) resulted in a significant increase of the RAR Bmax 10 h after 2DG application, with the Ka remaining unchanged. Shorter intervals, 1 or 5 h after 2DG application were ineffective on both the RAR Bmax and Ka. In the 2DG-adapted rats (6 doses of 2 DG, 500 mg/kg; 1 dose/day), decapitated 24 h after the last 2DG dose, the RAR Bmax was found significantly higher when compared to control group of animals. No further effect of the next dose of 2DG to repeatedly injected rats on the RAR Bmax and Ka was observed in animals killed 5 h after the seventh dose of 2DG. 2DG-induced intracellular glucopenia markedly up-regulates RARs in liver, but does not change the affinity of the receptor. Thus, the effect of 2DG on RAR concentration in liver specifically differs from that of IMO or laparotomy.

Animals↗

PubMed2000

Insulin, catecholamines, glucose and antioxidant enzymes in oxidative damage during different loads in healthy humans.

Exercise, insulin-induced hypoglycemia and oral glucose loads (50 g and 100 g) were used to compare the production of malondialdehyde and the activity of antioxidant enzymes in healthy subjects. Twenty male volunteers participated in the study. Exercise consisted of three consecutive work loads on a bicycle ergometer of graded intensity (1.5, 2.0, and 2.5 W/kg, 6 min each). Hypoglycemia was induced by insulin (Actrapid MC Novo, 0.1 IU/kg, i.v.). Oral administration of 50 g and 100 g of glucose was given to elevate plasma glucose. The activity of superoxide dismutase (SOD) was determined in red blood cells, whereas glutathione peroxidase (GSH-Px) activity was measured in whole blood. The concentration of malondialdehyde (MDA) was determined by HPLC, catecholamines were assessed radioenzymatically and glucose was measured by the glucose-oxidase method. Exercise increased MDA concentrations, GSH-Px and SOD activities as well as plasma noradrenaline and adrenaline levels. Insulin hypoglycemia increased plasma adrenaline levels, but the concentrations of MDA and the activities of GSH-Px and SOD were decreased. Hyperglycemia increased plasma MDA concentrations, but the activities of GSH-Px and SOD were significantly higher after a larger dose of glucose only. Plasma catecholamines were unchanged. These results indicate that the transient increase of plasma catecholamine and insulin concentrations did not induce oxidative damage, while glucose already in the low dose was an important triggering factor for oxidative stress.

Adult↗