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The molecular mechanisms and role of purinergic signaling in hypothalamus
Ivetic, Milorad ; Zemková, Hana (advisor) ; Mysliveček, Jaromír (referee) ; Smejkalová, Terézia (referee)
(EN): Extracellular ATP and purinergic P2 receptors (P2X and P2Y) are involved in a signaling network called "purinergic signaling" which is widely exploited in both somatic and neuronal tissues, and is also operative in endocrine system. The main focus of my thesis is on the role and expression of P2X and P2Y receptors in hypothalamic supraoptic nuclei (SON) producing hormones vasopressin and oxytocin, and the suprachiasmatic nuclei (SCN), the principal circadian pacemaker in mammals. In the first part of my thesis, we tested the hypothesis that P2X and P2Y receptors play a role in the enhanced release of hormones from magnocellular SON neurons stimulated through fasting/refeeding experimental protocol. We studied the effect of 2 h of refeeding after 48 h of fasting on hormone, P2X and P2Y mRNA expression in the SON tissue of 30-day-old rats, and the effect of ATP on electrophysiological properties of SON neurons in brain slices from control and fasted/refed rats. Quantitative real-time PCR revealed that the expression of P2X2 and arginine-vasopressin (AVP) mRNA was increased and P2Y1 mRNA expression was decreased in fasted/refed rats compared to controls, whereas P2X4, P2X7, P2Y2 and oxytocin mRNA levels were not significantly changed. Whole-cell patch clamp recordings showed that the amplitude...
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Neuropeptide Y and food intake regulation
Váchová, Barbora ; Nedvídková, Jara (advisor) ; Telenský, Petr (referee)
Neuropeptide Y (NPY) is a 36-amino acid member of NPY family. Its biosynthesis occurs in the hypothalamus and its expression also in adipose tissue was described in 2007. NPY is an ubiquitous hormone with central and peripheral effects which lead to the maintenance of homeostasis; NPY is also one of the most significant orexigenic hormones. NPY utilizes G-protein coupled receptors Y1, Y2 and Y5 for its food intake regulation; owing to serious side effects of currently used anti-obesity drugs, the ligands of these receptors (Y1R and Y5R antagonists and Y2R agonists) are considered as targeting molecules for a new therapeutic strategy for obesity treatment. This work is focused on the characteristic of NPY as an orexigenic agent and the possibility of using NPY-based drugs for obesity treatment.
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Ca2+ signalling in magnocellular neurones of the rat supraoptic nucleus.
Kortus, Štěpán ; Zápotocký, Martin (advisor) ; Šuta, Daniel (referee) ; Hromádka, Tomáš (referee)
The magnocellular neurosecretory cells (MNCs) of the hypothalamus project axons from the supraoptic nucleus to the posterior pituitary gland, where they secrete either oxytocin or vasopressin into the circulation. Oxytocin is important for delivery at birth and is essential for milk ejection during suckling. Vasopressin primarily promotes water reabsorption in the kidney to maintain body fluid balance. The profile of oxytocin and vasopressin secretion is principally determined by the pattern of action potentials initiated at the cell bodies in the hypothalamus. MNCs principally secrete hormones from terminals in the pituitary, but the secretion also occurs from their dendrites in the supraoptic nucleus, where they diffuse and affect the neighbouring cells. Mechanisms controlling the oxytocin and vasopressin secretion from MNCs have been extensively studied over the last decades and it is assumed that the relationship between Ca2+ signalling, secretion from dendrites, and the firing patterns is essential in understanding the magnocellular neurosecretory system. In this project, we combine mathematical analysis and experimental measurements of Ca2+ activity of MNCs of transgenic rats expressing an arginine vasopressin-enhanced green fluorescent protein (AVP-eGFP) or oxytocin-monomeric red fluorescent...
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The effect of stress on regulation and regeneration of glucocorticoids in animal models differing in response of hypothalamo-pituitary-adrenal axis
Vodička, Martin ; Pácha, Jiří (advisor) ; Svoboda, Jan (referee) ; Bendová, Zdeňka (referee)
Stress reaction is usually activated by the brain, when homeostasis is or perceived to be threatened. The stress signals are transmitted from the brain by two main branches; the sympathoadrenomedullary and the hypothalamo-pituitary-adrenal (HPA) axes and employ neural, humoral and immune pathways to cope with the stressor. Because of its potency, the stress reaction has to be precisely regulated. The HPA axis is regulated by feedback loops where its end product, corticosterone in laboratory rat and mouse, inhibits its activity. The effect of corticosterone does not depend only on the concentration of corticosterone but also on local metabolism of glucocorticoids via oxo-reduction catalyzed by the enzyme 11β -hydroxysteroid dehydrogenase 1 (encoded by the Hsd11b1 gene), which intracellularly regenerates active corticosterone from inactive 11-dehydrocorticosterone, or by extra-adrenal de novo steroidogenesis of glucocorticoids. We focused on analysis of stress response in experimental animals differing in HPA axis responsivity (Fischer 344 rats (F344) vs. Lewis rats (LEW) and germ-free (GF) vs. specific pathogen free mice (SPF)) with special emphasis on regulation of stress response, glucocorticoid regeneration and influence of gut microbiota. We found that stress modulated local regeneration of...
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Ca2+ signalling in magnocellular neurones of the rat supraoptic nucleus.
Kortus, Štěpán ; Zápotocký, Martin (advisor) ; Šuta, Daniel (referee) ; Hromádka, Tomáš (referee)
The magnocellular neurosecretory cells (MNCs) of the hypothalamus project axons from the supraoptic nucleus to the posterior pituitary gland, where they secrete either oxytocin or vasopressin into the circulation. Oxytocin is important for delivery at birth and is essential for milk ejection during suckling. Vasopressin primarily promotes water reabsorption in the kidney to maintain body fluid balance. The profile of oxytocin and vasopressin secretion is principally determined by the pattern of action potentials initiated at the cell bodies in the hypothalamus. MNCs principally secrete hormones from terminals in the pituitary, but the secretion also occurs from their dendrites in the supraoptic nucleus, where they diffuse and affect the neighbouring cells. Mechanisms controlling the oxytocin and vasopressin secretion from MNCs have been extensively studied over the last decades and it is assumed that the relationship between Ca2+ signalling, secretion from dendrites, and the firing patterns is essential in understanding the magnocellular neurosecretory system. In this project, we combine mathematical analysis and experimental measurements of Ca2+ activity of MNCs of transgenic rats expressing an arginine vasopressin-enhanced green fluorescent protein (AVP-eGFP) or oxytocin-monomeric red fluorescent...
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Circadian regulation of STAT3 protein in the SCN and it's activation by leptin in the SCN, other parts of hypothalamus and the pineal gland
Moníková, Veronika ; Bendová, Zdeňka (advisor) ; Jelínková, Dana (referee)
JAK/STAT signaling pathway is one of the most studied intracellular cascades transmitting signals from the extracellular environment to the cell nucleus in order to affect expression of target genes. Circadian clocks localized in the suprachiasmatic nuclei (SCN) of the hypothalamus are sensitive especially to light but they can respond to non-photic stimuli such as growth factors, opioids, leptin and cytokines that have been demonstrated to perform its function via the JAK/STAT signaling pathway. The recent findings of our laboratory demonstrated that STAT3 protein is highly produced by SCN of rat. Primary aim of our experiments was to test the circadian regulation of STAT3 production in SCN and describe the effect of exogenously administered leptin on STAT3 phosphorylation in the SCN, pineal gland and hypothalamic structures responsible for regulated feeding behavior and energy metabolism. Because activation of leptin receptors may stimulate a number of other signaling cascades, we chose phosphorylated forms of kinase ERK1/2 and GSK-3β as other markers of intracellular changes after administration of leptin in the studied structures. Our results proved rhythmic production of STAT3 protein in SCN of rat and indicated circadian regulation of sensitivity to leptin in hypothalamic structures. The data...
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Neuropeptide Y and food intake regulation
Váchová, Barbora ; Telenský, Petr (referee) ; Nedvídková, Jara (advisor)
Neuropeptide Y (NPY) is a 36-amino acid member of NPY family. Its biosynthesis occurs in the hypothalamus and its expression also in adipose tissue was described in 2007. NPY is an ubiquitous hormone with central and peripheral effects which lead to the maintenance of homeostasis; NPY is also one of the most significant orexigenic hormones. NPY utilizes G-protein coupled receptors Y1, Y2 and Y5 for its food intake regulation; owing to serious side effects of currently used anti-obesity drugs, the ligands of these receptors (Y1R and Y5R antagonists and Y2R agonists) are considered as targeting molecules for a new therapeutic strategy for obesity treatment. This work is focused on the characteristic of NPY as an orexigenic agent and the possibility of using NPY-based drugs for obesity treatment.
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