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A transcriptomic-based comparison of barley cultivars differing with respect to their low temperature acclimation capacity
Janská, Anna ; Ovesná, Jaroslava (advisor) ; Vaňková, Radomíra (referee) ; Honys, David (referee)
The PhD thesis is focused on a transcriptomics-based comparison of barley cultivars differing with respect to their low temperature acclimation capacity, with a particular focus on genes transcribed in the leaf and crown. The crown was of interest because of its importance for the winter survival of the plant. To involve both the first and the second phase of hardening, the test plants were exposed first to +3řC for 21 days, followed by - 3řC for one day. Freezing damage was assessed by measuring electrolyte leakage (Papers 2 and 3), using a modified version of a protocol developed by Prášil and Zámečník (1998). The same protocol was adapted to evaluate crown regrowth (Paper 2); for this purpose, the plants were cooled, then replanted and cut above the crown, and their survival rate calculated over the following week. Each RNA sample was queried by hybridization to an Affymetrix 22 K Barley1 GeneChip Genome Array (Close et al. 2004). The data were statistically analysed with the help of the software packages R, MAS 5.0 (Ihaka & Gentleman 1996) (Papers 2 and 3), Gene Spring GX 7.3 (Agilent Technologies, Santa Clara CA) and MapMan (Thimm et al. 2004; Usadel et al. 2005) (Paper 2), the "Self-Organizing Maps" algorithm (Kohonen et al. 1996) (Paper 3) and MIPS FunCat (Ruepp et al. 2004) (Paper 2). Paper...
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The role of PGC-1α during the development of a cardioprotective phenotype in a rat adapted to mild cold
Bajsová, Barbora ; Žurmanová, Jitka (advisor) ; Szeiffová Bačová, Barbara (referee)
Cold exposure elicits a thermoregulatory response in an organism. If the cold stimulus is sufficient, there is increased heat generation by shivering thermogenesis. Under prolonged action of the cold stimulus, shivering thermogenesis is replaced by non-shivering thermogenesis. Non-shivering thermogenesis in the rat takes place predominantly in brown adipose tissue (BAT), where the uncoupling protein 1 (UCP1) is activated in mitochondria under the control of adrenergic signaling. The activity of UCP1 is crucial for heat production. The coactivator PGC-1α plays an important role in mitochondrial biogenesis and in the energy metabolism of BAT and heart, and its function is associated with cardioprotection. Recently, the cardioprotective effects of mild cold adaptation at 8±1 řC were discovered in our laboratory, which reduced the size of ischemia-reperfusion injury without negative side effects. However, the molecular nature of the events induced by adaptation to mild cold is unknown neither in BAT nor in the heart itself. Therefore, the aims of the thesis were 1) to characterize the development of BAT at the level of expression of selected proteins and mitochondrial markers during acute cold exposure and chronic cold adaptation with subsequent recovery and 2) to determine the role of PGC-1α and...
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The effect of chronic cold on Ca2+-ATPase (SERCA2) in rat heart
Šeovićová, Maja ; Horníková, Daniela (advisor) ; Nováková, Olga (referee)
Acute cold exposure is a significant stressor activating heat production by shivering after the prolonged exposure cellular oxidative stress increases. Chronic exposure to cold lasting at least 2 weeks leads to the development of cold acclimatization. The main thermogenic role is taken over by non-shivering thermogenesis taking place in brown adipose tissue, which significantly increases its weight due to cold. Cardiac hypertrophy, hypertension and impaired renal function are frequently observed pathologies of acclimatization at 4-5 řC. Our laboratory recently introduced a model of mild chronic cold acclimatization at 8 řC, during which no damage to the heart or kidneys occurs and has proven cardioprotective effect on reducing infarct size. Hence, the influence of this cold acclimatization model on the other cellular and molecular processes needs to be investigated. The cardioprotective effect of cold acclimatization includes changes in β-AR signaling, activation of anti-apoptotic pathways or augmentation of the antioxidant system. The aim of this thesis was to investigate the effect of cold acclimation and subsequent reacclimation on proteins regulating Ca2+ levels in the rat heart (SERCA2 and phospholamban) and on the stimulation of regulatory proteins β-arrestin 1/2 and protein kinase PDPK1. The...
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The role of beta1 adrenergic receptors in heart of cold acclimated rat
Liptáková, Andrea ; Žurmanová, Jitka (advisor) ; Nováková, Olga (referee)
During cold acclimation the heat production is shifted from shivering to non-shivering thermogenesis, which is mediated by adrenergic signaling. It has also been observed, that cold acclimation may increase the organismal resistence to pathological stimuli and may affect functional parameters of cardiovascular system. However, acute exposure to sever cold is often associated with detrimental effects on the body. We have recently shown that chronic exposure to cold increases the heart's resistance to ischemia-reperfusion injury without negative side effects when mild temperatures are used, however the mechanism of protection is not yet known. The aim of this work was to determine whether: i) if the sensitivity of the heart to ischemia changes already after the first day of cold exposure and does not show any negative effects, ii) if β1-adrenergic signaling plays a role in chronic regimen of cold-induced cardioprotection. The results of this work showed that i) one day of exposure to mild cold did not change the sensitivity of the heart to ischemia and ii) metoprolol treatment reduced the infarct size in the control group, but did not affect the heart of cold-adapted rats. Key words : Heart, rat, beta 1 adrenergic receptors, cold acclimation
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The effect of cold exposure on beta-adrenegic signaling in the rat myocardium
Podojilová, Anna ; Novotný, Jiří (advisor) ; Ujčíková, Hana (referee)
and keywords The aim of this work was to evaluate the effect on rat myocardial β-adrenergic system of short-term (10 hours and 3 days) and long-term (5 weeks) cold exposure of rats with possible subsequent two-week recovery at normal temperature. The subtypes of β-adrenergic receptors (β-AR), their cognate G-proteins and adenylate cyclase have been characterized. β-AR are important components of cardiac regulatory mechanisms. They are involved in stimulating G-protein (Gs) and adenylate cyclase to increase cardiac contractility and frequency during stressful situations, including cold exposure. Heart tissue contains all three β-AR subtypes (β1-AR, β2-AR, β3-AR). While β1-AR couples with only Gs, β2-AR and β3-AR interact with the inhibitory G-protein (Gi). Electrophoresis and Western blotting showed a significant increase in β1-AR after a three-day exposure to cold. There was also a significant increase in β3-AR concentrations after a five-week cold exposure and this increase lasted for two weeks. There were no significant changes in the amnounts of β-AR cognate G-proteins (Gαs, Gβ, Gαi1/2 a Gαi3). In contrast, expression of adenylyl cyclase isoform V and VI significantly decreased during short-term exposure to cold. Using the saturation experiment with the β-antagonist [3H]CGP 12177, β-AR were...
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Mitochondrial respiration at cold acclimated rats. Comparison of tissues.
Flégrová, Eliška ; Žurmanová, Jitka (advisor) ; Nováková, Olga (referee)
Acclimation to cold or hardening is known for many decades through its beneficial effects on human health. In contrast, sudden exposure to cold, cold shock, is a great risk of cerebral and cardiac injury, especially in the elderly. There is very little published data on the cellular and molecular mechanisms induced by cold adaptation in heart and brain. The aim of this work was to describe and compare different properties heart, liver, brain and brown adipose tissue mitochondria of rats housed at 25 ± 1 řC and at mild cold (9 ± 1 řC, 5 weeks). The high-resolution oxygraphy, spectrophotometry and Western blotting analyses were used. We found differences in the respiratory control between the heart and liver. Cold acclimation decreased activity of the Krebs cycle enzymes. Fatty acid contribution to the respiration reached the maximum in brown fat and the minimum in the hippocampus. However, further study is necessary.
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Ischemia-reperfusion injury in cold acclimated rats
Vebr, Pavel ; Žurmanová, Jitka (advisor) ; Alánová, Petra (referee)
The effect of cold acclimation on body of mammals has been studied for many decades by using relatively low temperatures for acclimation (6-10 řC). The results of these experiments have shown the important role of the adrenergic and thyroid system during acclimation and negative impact on renal system at the same time. In contrast, a recent study on winter swimmers suggests a possibility of positive influence of hardening on cardiovascular system. There is no available study investigating a relationship between cold adaptation and ischemia-reperfusion injury. The aim of this study was to establish a protocol of isolated rat heart and its fixation at our workplace. Furthermore, to find the impact of mild cold acclimation on the ischemia-reperfusion injury of rat. Methods of ex vivo heart perfusion and fixation were successfully established. The effect of 5 weeks long cold acclimation in 10 ± 2 řC on left ventricle ischemia-reperfusion injury was observed. Powered by TCPDF (www.tcpdf.org)
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The role of the adrenergic system in the process of cold acclimation
Dynybylová, Anna ; Novotný, Jiří (advisor) ; Brabcová, Iveta (referee)
The ability to keep constant body temperature is necessary for mammals. There are effective strategies developed during the evolution - shivering and nonshivering thermogenesis, which start during cold exposure. This thesis is focused on the nonshivering thermogenesis which occurs in brown adipose tissue and is controlled by norepinephrine released from the sympathetic nerves if the cold stress persists for a long time. The principle of the heat production is based on function of the uncoupling protein 1 which disrupts the proton gradient in mitochondria thereby releasing heat the waste product. Similar mechanism of energy dissipation is used not only during cold exposure but also after digestion of high fat diet, which may provide clues for potential therapeutic treatment of obesity and associated metabolic diseases. This study summarizes current knowledge about the role of adrenergic signalling in the process of cold acclimation. Key words: cold acclimation, nonshivering thermogenesis, brown adipose tissue, sympathetic nerve system, uncoupling protein 1 (UCP1), obesity
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A transcriptomic-based comparison of barley cultivars differing with respect to their low temperature acclimation capacity
Janská, Anna ; Ovesná, Jaroslava (advisor) ; Vaňková, Radomíra (referee) ; Honys, David (referee)
The PhD thesis is focused on a transcriptomics-based comparison of barley cultivars differing with respect to their low temperature acclimation capacity, with a particular focus on genes transcribed in the leaf and crown. The crown was of interest because of its importance for the winter survival of the plant. To involve both the first and the second phase of hardening, the test plants were exposed first to +3řC for 21 days, followed by - 3řC for one day. Freezing damage was assessed by measuring electrolyte leakage (Papers 2 and 3), using a modified version of a protocol developed by Prášil and Zámečník (1998). The same protocol was adapted to evaluate crown regrowth (Paper 2); for this purpose, the plants were cooled, then replanted and cut above the crown, and their survival rate calculated over the following week. Each RNA sample was queried by hybridization to an Affymetrix 22 K Barley1 GeneChip Genome Array (Close et al. 2004). The data were statistically analysed with the help of the software packages R, MAS 5.0 (Ihaka & Gentleman 1996) (Papers 2 and 3), Gene Spring GX 7.3 (Agilent Technologies, Santa Clara CA) and MapMan (Thimm et al. 2004; Usadel et al. 2005) (Paper 2), the "Self-Organizing Maps" algorithm (Kohonen et al. 1996) (Paper 3) and MIPS FunCat (Ruepp et al. 2004) (Paper 2). Paper...
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Cold hardiness of larvae of the fruit fly, \kur{Drosophila melanogaster} (Diptera: Drosophilidae)
KORBELOVÁ, Jaroslava
We assessed survival of larvae of the fruit fly, Drosophila melanogaster at low temperatures (0°C and 5°C) after rearing them under seven different acclimation regimes. Larvae that have developed in a standard diet at 25°C showed 50% mortality after 12,6 min of the exposure to 0°C (Lt50 = 0.21 h). In contrast, larvae that have developed in a diet enriched with glycerol at 15°C, and were cold acclimated at 5°C during last two days of their development, had Lt50 = 38.6 h. It means that it was possible to increase the Lt50 at 0°C more than 180-fold using simple manipulations with rearing temperatures and diet composition. The physiological differences in duration of larval development, fresh mass, dry mass, hydration and total contents of proteins, lipids and glycogen between the larvae belonging to different acclimation variants are described. The samples for future detailed metabolomic analysis were prepared.
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