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Hippocampus Dysfunction in Quinpirole Sensitization Model of Obsessive-Compulsive Disorder
Brožka, Hana ; Stuchlík, Aleš (advisor) ; Jendelová, Pavla (referee) ; Kelemen, Eduard (referee)
Obsessive-compulsive disorder (OCD) is a serious psychiatric condition manifested by repeated thoughts followed by stereotypic compulsive behavior. Alterations to cortico-thalamo-striato- cortical circuits are most often implicated in the pathophysiology of OCD. However, many studies have also found a changed volume, shape and activity of the hippocampus in OCD patients. This work focused on the activity of hippocampal CA1 cells during stereotypical checking behavior and on cognitive flexibility in a quinpirole (QNP) sensitization model of OCD. The activity of CA1 hippocampal cells during stereotypical checking was assessed in an enriched open-field test in QNP sensitized rats. Arc+ (activity-regulated cytoskeletal associated protein, or Arg 3.1) mRNA expression profiles were determined in CA1 coronal hippocampal sections following stereotypical checking. After the establishment of stereotypical checking (10 sessions), rats were exposed to the arena and sacrificed after 5 minutes. QNP sensitized animals visited the same objects with the same frequency as during previous sessions, while control rats did not. Locomotor activity was comparable between QNP treated rats and controls. Following sacrifice, rat brains were flash frozen and sliced to 20 µm thick sections. Sections, mounted on slides, were hybridized...
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The effect of immunosupression on cell therapy in mouse model of Alzeimer's disease
Gajdoš, Roman ; Jendelová, Pavla (advisor) ; Chmelová, Martina (referee)
Alzheimer's disease is a chronic, progressive, neurodegenerative disease. It belongs to the most common type of dementia and worldwide it is statistically the fifth cause of mortality. The most common morphological markers are insoluble β amyloid plaques, hyperphosforylated tau proteins and formation of neurofibrilar tangles. Among the manifestations of the disease is amyloid angiopathy, synaptic transmission disorders and subsequent apoptosis, deterioration of cognitive functions and brain atrophy. Studies have shown that administration of mesenchymal stem cells (MSC) has an immunomodulatory effects and it can reduce the production and storage of β amyloid and thus improve cognitive functions. In preclinical studies, which are conducted in transgenic mice and often use xenografts, administration of immunosuppresion may lead to variety of positive or negative effects which can affect the results of the experiment. The subject of the master's thesis was to determine the effect of immunosuppression on experimental therapy with MSC in various time windows of AD progression (model 3xTg). At which scale and combination of immunosupression will influence the cell therapy's effects, the length of graft survival, mortality of experimental animals and changes at the cellular level. We have also assessed...
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The use of stem cells in the treatment of Amyotrophic lateral sclerosis
Řehořová, Monika ; Jendelová, Pavla (advisor) ; Klíma, Jiří (referee) ; Jiruška, Přemysl (referee)
Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease caused by degradation of motoneurons (MN). No effective treatment is currently available. Cellular therapy is considered to be a promising experimental treatment that could target the pathology of complex disease through many potential mechanisms. We compared the effect of three types of repeated applications of human mesenchymal stem cells (hMSC): intrathecal administrations, intramuscular administrations and the combination of these applications. Best results were obtained after combined repeated hMSC administrations. We observed the rescue of MN, neuromuscular junctions and decreasing levels of proteins involved in the signaling of necroptosis (Rip1, cl-casp 8), apoptosis (cl-casp 9) and autophagy (beclin 1), decreasing astrogliosis and the level of astrocytic connexin 43. Neural precursors derived from induced pluripotent stem cells (NP-iPS) are considered as other promising candidates for ALS therapy. Intraspinal administration of NP-iPS increased mRNA expression of BDNF and IGF-1, on the other hand decreased expression of proapoptotic casp 3. We also observed their effect on expression of components of perineural nets (PNN). Human embryonic stem cells (hESC) are the other cell candidates for the treatment of neurodegenerative...
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Treatment of spinal cord injury targetting secondary mechanisms
Valášková, Barbora ; Jendelová, Pavla (advisor) ; Hájek, Milan (referee) ; Rokyta, Richard (referee)
1 Abstract Traumatic spinal cord injury (SCI) is severe condition with devastating long- lasting consequences. There is still no available treatment. After initialing mechanical trauma, there is a huge cascade of secondary reactions which amplifies the damage. This thesis is focused on potential therapeutic effect of photobiomodulation, natural anti- inflammatory compounds epigallocatechin-3-gallate (EGCG), curcumin, their combination and extrapure synthetic form of curcumin called "nanocurcumin" on impacts of experimental model of SCI in rats. Photobiomodulation using combination of two synchronized wavelengths 808 and 905 nm improved functional recovery after SCI evaluated by battery of locomotor tests and somatosensory plantar test. The group treated by photobiomodulation obtain better results in all tests. The histopathological analysis showed a positive effect on white and gray matter sparing and our data suggests an upregulation of M2 microglia/macrophages in photobiomodulation treated rats assessed by immunohistochemical and RT-qPCR analysis. Our results demonstrated that the photobiomodulation is a promising non-invasive therapy for improving functional recovery and tissue sparing after SCI. EGCG and curcumin are natural compounds known in Chinese medicine for centuries. Their neuroprotective and...
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Modulation of memory using enzymatic digestion of perineuronal nets in perirhinal cortex
Procházková, Natálie ; Jendelová, Pavla (advisor) ; Chmelová, Martina (referee)
In the adult brain, neuronal plasticity is regulated by a specialized structure of extracellular matrix, the perineuronal nets (PNNs), which restrict synaptic plasticity by binding molecules of inhibitory nature and posing as a physical barrier to alterations in neuronal connectivity. This effect is abolished by removal of PNNs by the enzyme chondroitinase ABC (chABC), which enables to reopen critical period window and leads to memory improvement. Here, we utilized chABC and a novel approach in removing the PNNs in perirhinal cortex, using the enzyme hyaluronidase (Hyase), to assess differences in the use of these enzymes in object recognition (OR) memory improvement and alterations in the structure of neuronal network of wild type mice. Our findings suggest that Hyase may be a more convenient tool to PNN removal than chABC, as Hyase surpasses chABC in promoting the OR memory, influence larger portion of neuronal network by affecting both inhibitory and excitatory neurons, and provides extended temporal window for experimental modulation of activity-dependent synaptic plasticity. Key words: perineuronal nets, synaptic plasticity, chondroitinase ABC, hyaluronidase, perirhinal cortex, memory
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Cellular and molecular basis of a skin regeneration in amphibians and mammals
Hybešová, Michaela ; Krylov, Vladimír (advisor) ; Jendelová, Pavla (referee)
Wound healing of skin in mammals and its regeneration in Amphibians are crucial biomedicine topics in the last few decades. The most important aspect in humans is the scrarring proccess and the effort to substitute it with the regeneration producing functional and differentiated tissues. To modulate the formation of scar it is neccessary to compare both proccesses. The core animal model is axolotl (Caudata) where regeneration takes place during whole lifespam. On the other hand, in frogs (Anurans) this phenomenon is restricted up to metamorphosis. After metamorphosis, the immune system of Anurans is similar if compared to mammals. Similarly, the transition from early embryonic development in mammals, where the fetus was able to completely regenerate damaged tissue to an adult type of healing, goes hand-in-hand with the development of the immune system and structural differentiation of damaged tissue. Thus, the inflammatory cells and their regulation, the formation of ECM, which includes fibroblast proliferation, and the production of appropriate cytokines are key factors that distinguish the process of healing with and without scarring.
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Effect of iron oxide nanoparticles with ascorbic acid on neural stem cells
Jiráková, Klára ; Moskvin, Maksym ; Horák, Daniel ; Jendelová, Pavla
Cells labelled with iron oxide nanoparticles (ION) can be tracked by magnetic resonance imaging (MRI) in several applications. However, various studies demonstrated toxicity and oxidative stress induction associated with nanoparticles exposure. We analysed biologic effects after the exposure of two types of iron oxide nanoparticles (with and without an antioxidative agent, an ascorbic acid) on human neural stem cells. The labelled cells in gel phantoms were detected in MRI and they showed decreased relaxation rates in comparison with control. ION slightly decreased cell proliferation in comparison with unlabelled cells, which was dependent on concentration and presence of ascorbic acid. None of the nanoparticle type showed negative effect on cell viability and both demonstrated minor effect on reactive oxygen species (ROS) formation. Unfortunately, ascorbic acid bound to nanoparticles did not show any effect on ROS attenuation. Cells exposed to both types of nanoparticles showed increased positivity for a phosphorylated form of H2AX a marker of double strand breaks. We showed that ION in low concentrations do not affect cell viability, but have negative effect on cells on DNA level. Their potential use for oxidative stress reduction is dependent on the concentration of ascorbic acid bound to the nanoparticles and this should be further increased.
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Anti-inflammatory compounds and stem cells in treatment of spinal cord injury
Kárová, Kristýna ; Jendelová, Pavla (advisor) ; Sameš, Martin (referee) ; Chvátal, Alexandr (referee)
Despite intense scientific efforts, spinal cord injury (SCI) remains to be a severe neurological condition that has no treatment. Currently, therapy is based on alleviating pressure by surgical spinal cord decompression, administration of methylprednisolone and physical therapy. In this study, therapeutic effects of anti-inflammatory compounds and of three types of stem cells were tested in a balloon compression model of spinal cord injury in rats. Natural compounds epigallocatechin-3-gallate (EGCG) or curcumin were administered in situ and then intraperitoneally every day for up to 28 days. Human bone marrow mesenchymal stem cells (MSCs), human spinal neural precursors (SPC-01) and neural precursors derived from human induced pluripotent stem cells (iPS-NPs) were transplanted intrathecally (MSCs) or via spinal injection into immunosuppressed rats 7 days after induction of SCI. To determine effects of therapies, changes in motor function was tested by open field test BBB, flat beam test and score, Plantar test and rotarod. Morphometric analysis was used to assess gray/matter sparing and cavity size. Immunohistochemistry was used to determine survival and differentiation of transplanted cells, activation of classical pathway of NFκB (p65 nuclear translocation), astroglial activation (GFAP) and...
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Cellular Reprogramming as a Tool for Harvesting Patient-specific Stem Cells
Pisal, Rishikaysh ; Mokrý, Jaroslav (advisor) ; Hampl, Aleš (referee) ; Jendelová, Pavla (referee)
Cellular reprogramming as a tool for harvesting patient specific stem cells In the year 2006, Dr. Yamanaka surprised the entire field of medicine, by reporting a technique of inducing pluripotency in somatic cells. In his article, he had displayed that fibroblasts could be reprogrammed to pluripotent stem cell state, by ectopic expression of four transcription factors namely OCT4, SOX2, c-MYC and KLF4. His discovery made a paradigm shift in the field of reprogramming because previous methods of reprogramming were dependent on use of human oocytes and this raised ethical concerns. Moreover, his technique of cellular reprograming broadened the spectrum of application of somatic cells in regenerative medicine. Objectives of my research were focused on; development of an optimised protocol for detection of mycoplasma that commonly infects animal tissue culture; detailed characterization of reprogrammed clones; targeted differentiation of iPSC towards myogenic lineage, and construction of an expression vector, optimised for miRNA expression. For detecting mycoplasma infection, we adapted the protocol of Uphoff et al. (2002). By skipping the DNA extraction step (reported in the original protocol) and instead directly using cell culture supernatant and a robust polymerase enzyme for performing PCR, we...
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