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Immunohistochemical analysis of the expression of selected signaling molecules in rat brain: the effect of morphine withdrawal
Přítulová, Eliška ; Novotný, Jiří (advisor) ; Mrózková, Petra (referee)
Morphine is one of the most commonly used analgesics for pain, but its clinical use may be accompanied by the development of tolerance and dependence. Abuse of morphine can then lead to the development of severe withdrawal symptoms. Given the knowledge gathered so far, morphine addiction research often examines the impact on areas of the brain involved in the reward system. The main goal of this work was to investigate the effect of long-term administration of morphine and morphine withdrawal on certain signaling molecules that are related to the molecular action of opioids in selected areas of the rat brain. We focused on the cAMP response element-binding protein (CREB) and its active phosphorylated form (pCREB), as well as on G-protein signaling regulator RGS4. Our results indicated that morphine administration may cause a decrease in CREB expression in the basolateral amygdala in morphine-affected rats. We also found a reduction in CREB phosphorylation in the CA1 region of the hippocampus, possibly due to morphine withdrawal for three months. In this study, we did not observe any statistically significant changes in RGS4 expression in the prefrontal cortex and hippocampus due to morphine administration and subsequent morphine withdrawal. Key words: morphine, dependence, withdrawal, rat brain,...
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The role of miRNA in injury and regeneration of spinal cord tissue
Šprincl, Vojtěch ; Romanyuk, Natalyia (advisor) ; Mrózková, Petra (referee)
MicroRNAs are small non-coding RNA molecules of a length about 20-24 nucleotides, that regulate gene expression post-transcriptionally. They interfere mRNA molecules via base- pairing with complementary sequences. Recently it was shown that they play an important role in injury and regeneration of nervous tissue. The aim of this bachelor thesis is to describe possible role of miRNAs in central nervous system injury with focus on spinal cord injury.
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Thermally gated TRP channels in nociceptive neurones
Chvojka, Štěpán ; Vlachová, Viktorie (advisor) ; Mrózková, Petra (referee)
Transduction ion channels are gated in response to a variety of external stimuli and this process is critical for the proper functioning of sensory neurons. These specialized proteins enable the survival of any organism, which depends on having adequate information about the external environment. The thermosensitive TRP (transient receptor potential) ion channels, whose molecular structure has been identified during last decades, enable the transduction of thermal stimuli in primary nociceptive neurons. During the last decade, molecular biological techniques have provided new tools for studying the structure of these specialized transduction ion channels in relation to their function and to understand more deeply their physiological roles. The aim of this bachelor thesis is to give an overview of recent evidence regarding the functional and physiological properties of sensory-neuron specific mammalian TRP ion channels that are activated by thermal stimuli: heat and cold.
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Methods using preference in pain research
Komárková, Lucia ; Vaculín, Šimon (advisor) ; Mrózková, Petra (referee)
The thesis deals with neuropathic pain measurement in laboratory mice and rats. Neuropathic pain is characterized besides evoked allodynia and hyperalgesia also by spontaneous pain and anhedonia, further motivational-affective part of pain is highlighted. Traditional pain threshold testing is not appropriate method to measure these aspects of pain. Therefore, the aim of the thesis is to describe preference methods in pain research. We focused on three methods - conditioned place preference, sucrose preference test and thermal place preference. The first mentioned method creates an association between pain relief and a specific place, which afterwards the animal prefers. The conditioned place preference can be used to detect spontaneous pain. Sucrose preference test monitors decrease in consumption of sugar water after exposure to chronic pain, which can be interpreted as a sign of depression, so thanks to this method we have the opportunity to explore the interaction between pain and depression. The thermal place preference method studies changes of preferences between the cold and hot plate after induction of pain or analgesics. The findings based on preference methods help us to examine all aspects of neuropathic pain and develop more options for pain treatment.
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Experimental rat models in traumatic spinal cord injury research
Kloudová, Anna ; Machová Urdzíková, Lucia (advisor) ; Mrózková, Petra (referee)
A large number of spinal injuries that cause damage to the spinal cord occur every year. Such damage to spinal cord tissue leads to a number of dysfunctions that significantly reduce quality of life and therefore the development of effective therapies remains a very important issue. The use of animal models is considered the major tool of this research field, enabling the testing of new treatments as well as observing ongoing pathophysiological processes. Rats are the most frequently used animal model due to the many possible ways of creating a spinal cord lesion on them. The contusion model uses a computer-controlled device to simulate the spinal cord contusion and therefore is highly precise. However, a compression model seems to be the most suitable model for pathophysiological studying. Besides contusion it enables persistent compression to the spinal cord. Transection and hemisection methods are intended for axonal regeneration research, as only these models can reliably distinguish new growing fibers from the preserved ones. Less often photochemical and excitotoxic models are used. Lastly, dislocation and distraction of vertebrae or damage to the spinal cord using heat or cold can be carried out. Each of the models have their own unique features that make them optimal to use in different...
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Modulation of synaptic transmission, studies on spinal cord slices in vitro
Mrózková, Petra ; Paleček, Jiří (advisor) ; Krůšek, Jan (referee)
Modulation of a synaptic transmission in the spinal cord dorsal horn plays a key role in nociceptive signalling, especially in states of pathological pain. The goal of this study was to develop a method for calcium imaging in spinal cord slices in vitro. This method allowed us to record changes of intracellular free calcium ions concentration (iCa2+ ), that are a major mediator of neuronal plasticity. In this work, we have focused on application of this method in a conventional fluorescence microscope and on the role of different neuromodulators of synaptic activity. Changes of iCa2+ induced by dorsal root electrical stimulation were recorded altogether in 744 dorsal horn (lamina I and II) neurons. In the first series of experiments, stimulation protocols activating preferentially A and A + C dorsal root fibers were used and long-term stability of the calcium responses was verified. The dorsal root stimulation induced in the neurons fast and delayed type of calcium response. Application of AMPA and NMDA receptors antagonists, CNQX (50μM) and MK801 (45μM), reduced the calcium response amplitude and confirmed the importance of glutamate receptors in synaptic activation. In several experiments the effect of capsaicin a TRPV1 receptors agonist, application was tested. Application of even low...
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Effects of sulfated neurosteroids on proteins involved in excitatory synaptic transmission
Naimová, Žaneta ; Smejkalová, Terézia (advisor) ; Mrózková, Petra (referee)
The discovery of steroid compounds capable of synthesize or accumulation in CNS and PNS led to a question about their function. Neurosteroid compounds are capable of modulating synaptic transmission. Effect is direct and fast mediated through nongenomic mechanisms. They are known to affect wide array of channels and receptors - both excitatory and inhibitory. This thesis summarizes findings about effect of sulfated neurosteroid on proteins involved in excitatory synaptic transmission. Thesis covers findings about ionotropic glutamate receptors, TRP channels, metabotropic receptors, sodium and potassium channels. Excessive or insufficient activity of these proteins involved in synaptic transmission can lead to a pathological condition. The purpose of this thesis is to summarize findings about effect of these compounds, point out structural and function characteristic probably responsible for their action and to outline possible pharmacological usage.
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The role of biofluid biomarkers in the differential diagnosis of neurodegenerative diseases
Katonová, Alžbeta ; Čechová, Kateřina (advisor) ; Mrózková, Petra (referee)
Until recently, the diagnosis of Alzheimer's disease and other neurodegenerative diseases was dependent only on clinical expression, the definitive confirmation of which was possible only post- mortem. Currently, biofluid biomarkers represent a reliable source for the diagnosis of serious neurodegenerative diseases already during the patient's lifetime. The identification of reliable biomarkers is essential for increasing diagnostic accuracy, monitoring the clinical condition, more precise selection of individuals for clinical trials and the associated development of causal treatment. The aim of the bachelor's thesis was to conduct a thorough literature review focusing on clinically and experimentally used biological markers in the diagnosis of neurodegenerative diseases and their role in the pathophysiology of these diseases. Key words: Neurodegenerative diseases, biomarkers, proteinopathies, neurocognitive impairment, cognition
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Mechanisms of neurophatic pain states development
Přibáňová, Tereza ; Mrózková, Petra (advisor) ; Kuchtiak, Viktor (referee)
Pain is a natural warning signal that protects organisms from actual or potential damage. Upon the stimulation (burns, cuts, inflammation) of nerve endings - nociceptors, nerve signals are conducted via the peripheral nerve fibres into the spinal cord and brain, where they are then processed as painful, and a reaction occurs. Neuropathic pain, on the other hand, is pain caused by an injury or disease of the somatosensory system itself. Neuropathic pain has a substantial impact on the patient's quality of life and is likely to become more prevalent as the population grows older and the rates of diabetes and chemotherapy treatments rise. However, the treatment of neuropathic pain is often insufficient and comes with a number of undesirable side effects, which constitute a significant clinical problem. Research leading to the understanding of the molecular mechanisms of the development and maintenance of neuropathic pain is necessary in order to enhance the treatment of these states and to make it more effective. There is a myriad of factors responsible for the development of neuropathic pain, namely mechanisms which maintain the balance between inhibitory and excitatory somatosensory signalling, changes in the amount or composition of receptors and channels at the surface of the neuron, and most...
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