Suprachiasmatic nuclei (SCN) are primary center of mammalian circadian rhythms. To maintain a 24 hour period of its rhythms, SCN are synchronized with phase of external environment. Regular changes of light and darkness are known to be the main external synchronizer that determines the period of SCN rhythms. Information about light is being transferred from retina to the ventrolateral region of SCN through excitatory synapses where ionotropic glutamate receptors (iGluRs) play a primary role in the signal transduction. Posttranscriptional modifications of RNA can alter the functional properties of iGluRs, thus this process contributes to synaptic plasticity. The extent of posttranscriptional modifications of iGluRs can be in vitro affected by neuronal activity altered by pharmacological manipulation. The aim of this study was to determine possible changes of posttranscriptional modifications of iGluRs in in vivo rat SCN model and how this process can be regulated. RNA posttranscriptional modifications of GluA2 subunit of AMPA receptor (AMPAR) and GluK2 subunit of kainate receptor were assessed using PCR and subsequent sequencing of amplified DNA. Using quantitative PCR, we also determined mRNA expression of GluA1 and GluA2 subunits of AMPAR and the editing enzyme ADAR2 in SCN. Our results showed...

Our body is influenced by many cyclical changes in the environment, such as day and night or seasons. To predict these changes and react to them in time, the organism is equipped with inner clock, which rhythmically influences many physiological processes, such as sleep or metabolic rhythms. Disrupting our inner rhythms at molecular and behavioral levels contributes to many serious disorders. It is necessary that all mechanisms of the inner circadian clock are developed and set up properly. Circadian clocks are set up by the mother, who passes rhythmical information about day and night cycle on to her embryo. Though a great attention is devoted to revealing the nature of this synchronization between the mother and her pup, the mechanisms of this process have not been fully understood yet. The aim of this thesis is to contribute to actual understanding of this synchronization. Experiments, performed in this thesis, relate to studying the ability of maternal signals to synchronize embryos with the environment. Feeding and light regime of pregnant rats was manipulated and the effect of these changes on the neuronal activity within the suprachiasmatic nuclei of 19-day embryos was analyzed.

Hippocampus is one of the main components of the limbic system. It plays an important role in the process of memory formation and recent literature shows that, like in other peripheral organs and brain structures, there is a circadian clock present in the hippocampus. The aim of the thesis was to study the hippocampal circadian clock under various conditions. The first part of the thesis examines the influence of glucocorticoid hormones on the clock gene expression in the hippocampus. Glucocorticoids are mammalian steroid hormones secreted from the adrenal glands that affect many processes in the organism. The glucocorticoid secretion is under the control of the circadian system, causing their levels to exhibit a pronounced diurnal rhythm. The hippocampus provides feedback to the hypothalamo-pituitary-adrenal (HPA) axis and is thus involved in the regulation of glucocorticoid hormone secretion. Hippocampal neurons also express glucocorticoid hormone receptors (GR). This thesis explores the effect of the absence of glucocorticoid hormones on the clock gene expression in the hippocampus. The absence of glucocorticoids was due to adrenalectomy and its effect was further compared with the effect of endogenous glucocorticoid replacement by daily injections of the GR agonist dexamethasone, emulating...

Circadian clock in the suprachiasmatic nucleus of the hypothalamus (SCN) regulates daily rhythms in behavior and physiology and is an important part of the mechanisms regulating mammalian homeostasis. SCN are synchronized with a 24hour cycle mainly by light, but they can also be regulated by a variety of nonphotic signals, such as growth factors, opioids, cytokines, or lipopolysaccharide (LPS), which act by inducing the JAK/STAT signaling pathway. STAT family proteins (i.e. signal transducers and activator of transcription) regulate many aspects of cellular physiology, from growth and differentiation to immune response. However, the JAK/STAT signaling pathway has not yet been studied in the SCN and the function of STAT proteins in the SCN has not yet been clarified. In the first part of the thesis, we focused on localization of STAT3 and STAT5 proteins in the rat SCN and determination of rhythm in proteins and mRNA. Our experiments showed the daily rhythm in the levels of STAT3 protein in SCN astrocytes of rat with low but significant amplitude and with maximum in the morning. In addition, we revealed strong but nonrhythmic expression of STAT5A protein in astrocytes and STAT5B protein in nonastrocytic cells of SCN. It was also found that Stat3 mRNA show, similarly to protein, circadian rhythm in...

In this diplomat theses are considered the effects of light on human beings, especially on physical performance. The goal of this work was to design a suitable method for evaluation of the effects of light on physical performance. Furthermore, to verify designed methods in laboratory environment and the assessment of collected data. After creating this method several people were tested under different light conditions and collected date evaluated. The results are suggesting that high intensity lighting with low wave lengths leads to an increase of physiological activity in organism and therefore to a potential decreasing physical performance.

The circadian clock generates circadian rhythms, which participate on regulation of a number of signalling pathways. Disruption of the circadian regulatory mechanism is linked to a development and a progression of certain types of cancer including colorectal tumorigenesis. Progression of tumorigenesis depends on the cell cycle machinery related to cell proliferation and apoptosis. MiRNAs play a role in initiation and progression of tumorigenesis because they interfere in regulatory pathways associated with tumorigenesis. The aim of the thesis was to determinate existence of circadian rhytms in clock controlled genes (Tef, Dbp), miRNAs (miR-1-3p, miR-16-5p, miR-34a-5p, miR-155-5p, miR-192-3p) and genes of the cell cycle machinery (Ccnd1, Ccne1, Ccna1, Ccnb1) and apoptosis (Casp3, Bcl2, Bad). Further, to compare detected circadian rhythms during aging and neoplastic transformation of colon by quantitative RT-PCR. We have observed circadian expression of Tef, Dbp, Ccne1, Ccna1, Ccnb1, Casp3 and Bcl2 in young mice colon, Tef, Dbp, miR-1-3p, Ccne1, Ccna1 in old mice colon and Tef and Dbp in colorectal tumors. In summary, circadian expression of clock controlled genes varied but was maintained in mice colorectal tumors. In aging we demonstrated weakening of circadian rhythms of the genes of the cell...

The biological clock are responsible for entraining of circadian rhytms in many physiological and behavioral functions, including the activity of immune system. In mammals, the biological clock are located in the suprachiasmatic nuclei of the hypothalamus and also on the periphery. The main oscillator in suprachiamatic nuclei synchronizes the phases of the peripheral oscillators in tissues and the other brain areas and it also drives synthesis of melatonin in the pineal gland. Its endogenous period Tau (τ) is being synchronized with external 24-hour day under normal conditions, mostly by the light-dark cycles during the day and night and also by non-photic stimuli. By the application of endotoxin, disruption of normal endogenous conditions in organism causes activation of the immune response, behavioral changes and phase shifts of the circadian rhythmicity. This work examines effect of acute application endotoxin lipopolysaccharid on the cellular signaling cascades in the cells of suprachiasmatic nuclei and pineal gland. Our data indicate, that levels of phosphorylated forms of kinases GSK3β, ERK1/2 and levels of phosphorylated forms transcription factors STAT3 are substantially modificated after administration of lipopolysaccharid in the suprachiasmatic nuclei and especially in the pineal gland of...

The smallest free living eukaryote known as Ostreococcus tauri became along with some related species great experimental models for iron uptake research in marine picoplankton. The ecological context of such topic is very interesting considering the nature of adaptation to iron limitation and its connection to the copper metabolism. Our experiments has simulated iron and copper limiting conditions of the ocean, as a control we used iron and copper repleted cultivation media. The maximum cell counts were two- to threefold higher in iron-repleted medium compared to iron- depleted one. There was the only exception showing no difference in growth - O. lucimarinus coming from open ocean with high irradiance levels and very low iron concentrations, which all together made it the best adapted species. The reinoculation of cell cultures after a week cultivation into to the same iron/copper containing media led to unmasking the copper effect on growth, which was much weaker than encountered with iron (again except of O. lucimarinus). Iron sufficiency positively affects heme b and chlorophyll a and b content with no significant copper dependency. The circadian timing of heme b and chlorophyll a and b content shows the increasing trend during the day followed by decrease at night. This might be caused by...

The main mammalian circadian pacemaker stored in suprachiasmatic nuclei of the hypothalamus (SCN) is adapted to changes in the external environement by synchronization of its endogenous period with periodic changes of light and dark during day and night. The information about light travels via glutamatergic retinohypothalamic tract to the ventrolateral part of the SCN. Activation of ionotropic glutamate receptors in this area provably mediates the transfer of information about light on the transcriptional mechanism of light-sensitive cells. The role of the NMDA type of ionotropic glutamate receptors is well studied in this field and it is known that some NMDA receptor subunits show a circadian rhythm and an increased expression after a light pulse. Signalization via AMPA type receptors is much less elucidated. The aim of this thesis was to determine which AMPA receptor subunits are expressed in the SCN of the rat and if these subunits show a daily rhythm of expression and a reactivity to light pulse, as well as to outline the possible roles of distinct AMPA receptor subunits in the SCN. Keywords: circadian rhythms, suprachiasmatic nuclei, glutamate receptors, AMPA

Glucocorticoids are mammalian steroid hormones secreted from the adrenal gland. The basal levels of glucocorticoids show a pronounced diurnal rhythm with maximum at the beginning of the active period and minimum at its end. Glucocorticoids have an influence over a variety of metabolic functions and their secretion is tightly regulated. This regulation also depends on the circadian system, which utilizes glucocorticoids to entrain the peripheral tissues by inducing rhythmic gene expression. The mechanisms by which glucocorticoids influence mammalian circadian system has not yet been precisely defined, especially concerning the influence of glucocorticoid signalling on gene expression in different tissues and the dynamics of glucocorticoid receptor (GR) occupancy. This thesis studies the influence of ablation of glucocorticoid signalization induced by adrenalectomy on the clock gene expression of in the central clock in the suprachiasmatic nucleus and peripheral clocks in the hippocampus and distal colon. The effect of adrenalectomy on gene expression is compared with the effect of restricting the feeding time, which has also been shown to affect glucocorticoid levels in the body. Other experiments were aimed at elucidating impact of changing the activity of GR on gene expression using synthetic GR...