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Biological significance of neuroactive steroids in animal models of brain diseases
Rambousek, Lukáš ; Valeš, Karel (advisor) ; Votava, Martin (referee) ; Chvátal, Alexandr (referee)
Neurosteroids play an important role in the brain physiology and pathophysiology. They control inhibitory and excitatory neurotransmission. The presented thesis aims to investigate the biological significance of neuroactive steroid 3_5_-pregnanolone glutamate (3_5_P-Glu). We investigate the effect of 3_5_P-Glu in naïve rats. Next, we evaluate the effects of 3_5_P-Glu in animal model of schizophrenia and excitotoxic lesion of hippocampus induced by N-methyl- D-aspartate (NMDA). Furthermore, we morphologically characterize the NMDA lesion model. 3_5_P-Glu did not induce significant psychotomimetic side effects such as hyperlocomotion, sensorimotor grating deficit or memory impairment. Next, 3_5_P-Glu showed dose dependent pro-cognitive effects in animal model of schizophrenia; however, it had no effect on hyperlocomotion in this model. 3_5_P-Glu also ameliorated spatial learning deficit of rats induces by NMDA lesion of hippocampi in the Carousel maze and had mild effect on NMDA induced damage of hippocampus when applied before. Additionally, the morphological analysis of hippocampal NMDA lesion revealed overexpression of NMDA receptor NR1 and NR2B and downregulation of GABAA receptor _5 subunits. The lesion was very conservative, did not spread to other structures and did not affect GABAergic...
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Membrane properties of neural stem/progenitor cells during in vitro differentiation and after transplantation into the rat brain
Prajerová, Iva ; Chvátal, Alexandr (advisor) ; Vyklický, Ladislav (referee) ; Mazurová, Yvona (referee)
AOSTRACT The aim oflhis .....orl:. was to elucidate the dilTercntiation mechanisms of neural stcmlpmgenitor cells in vitro and their potentiallO survive and ditfcn:ntiate in \'il'o, al\er trarlSplnntatioo into thc injured mt conex. IImnunohistochcmistI)' "'lIS used for cell idcntiticatioos. and lhe p'0pCl ties ofK' aod Na' voltagc-gated ion ehnnnels were studiC<! using the pat,h-(:Iamp teclmique, We Ilave demonstrnted lhal inunortalised green lluort:SCCnll)r()lein (GFP)INE-4C neural stem cells derivtXI fmm tbc neurocpithelium of p5l'<lctieicm mouse cmbryos iII cmbryonic day (E)9 are ablc to dilferentiate i"to !leUrQns in \1"0, After tmnsplantalioo illto the site of a photlxhcmical lC'iion of aduh rol5, GFP!NE4C cdls slIn'ive and give Tise to neurons. astrocytt5 and oHgodmdrocytcs. Prima!)' embryonic ncura.l Slem cells were isoLaled frum O6IGFP ntice, in which GFP is expressed un(\cr tbc IXlfItrol of06. 11 promoter of thc mDachl gene. which is in\'Olved in tbe development of Ihc concx. At E1 2. D6 is specifically exprcssed in loc neurd.! stem cclLs of the (\(nal tclcnocephalon. from whieh cortical neurons arise, We have shown lila! D61GFP neurJI stem odLs isolated fmm 1312 embfyos are able to give rlse to neuroos and glial fibrillary acidic protcin (GFAP)-posilivc cel1s ill \'i"o nnd that after...
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The study of functional and molecular properties of NMDA receptors
Vyklický, Vojtěch ; Vyklický, Ladislav (advisor) ; Chvátal, Alexandr (referee) ; Tureček, Rostislav (referee)
N-methyl-D-aspartate (NMDA) receptors are a subtype of ionotropic glutamate receptors, which mediate fast synaptic transmission in the vertebrate central nervous system and play a key role in synaptic plasticity. NMDA receptor overexcitation leads to cell death that underlies many serious neurological and psychiatric disorders. The aim of our research was to characterize novel drugs that modulate NMDAR activity. We have tested a series of newly synthesized steroidal compounds for their activity on recombinant receptors. We have demonstrated a relationship between the structure of the analogues of a naturally occurring neurosteroid pregnanolone sulfate and their ability to modulate NMDA receptors. The results of our experiments characterize the role of substituents at the steroid A and D ring. We have found specific substituents on carbons C3 and C17 of the steroidal skeleton that lead to a substantial increase of steroid efficacy and the ability to positively affect the ratio between the inhibition of stationary and non-stationary receptor activation. These two states represent receptors long-term activated (tonically) and synaptically activated (phasically), respectively. Using electrophysiological and optical methods in combination with mathematical modeling we have shown that the plasma membrane plays an...
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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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The electroencephalographic changes in animal serotonergic model of psychosis-the study of EEG connectivity in relation to behavior
Tylš, Filip ; Páleníček, Tomáš (advisor) ; Mareš, Pavel (referee) ; Chvátal, Alexandr (referee)
The present thesis deals with the electrophysiological and behavioral correlates of the effects of serotonergic psychedelics in an animal model of psychosis. In the general part, we describe the phenomenology and neurobiology of psychotic diseases and the altered states of consciousness induced by serotonergic psychedelics. Furthermore, we outline the pharmacological models of acute psychosis with a focus on serotonergic substances, particularly the active substances from hallucinogenic mushroom - the indolamine psilocybin and its active metabolite psilocin. We briefly explain the method of quantitative electroencephalography (QEEG) and summarize the main findings of QEEG in animals and humans intoxicated by serotonergic psychedelics. The experimental part of the thesis describes the pharmacokinetics of psilocin, its effects on the behavioral pattern, locomotion and sensorimotor processing of information, as well as sex differences in the behavioral response. A medium dose of psilocin caused atypical behavioral figures, decreased locomotion and disturbance in sensorimotor processing of information, with the females being more resistant to this effect. Another focus of the thesis is the dynamics of QEEG changes in time (EEG absolute power spectra and EEG coherences), as recorded from the cortex of...
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Functional properties and plasticity of the rat auditory cortex
Pysanenko, Kateryna ; Tureček, Rostislav (advisor) ; Maršálek, Petr (referee) ; Chvátal, Alexandr (referee)
Auditory cortex (AC) is the ultimate target of afferent auditory pathways and plays a crucial role in the perception and localization of complex sounds. In the thesis were investigated and discussed three aspects of the AC function: i) diversity of the intrinsic passive and active electrical properties of core and belt AC neurons, ii) the modulatory function of the descending connections from the auditory cortex to the inferior colluculus and iii) the effect of a complex acoustical environment applied during the critical period on the responsiveness of auditory cortex neurons in rats. Within the AC can be distinguished two fundamental areas: centrally located core area and peripheral belt area. Recordings of sound-evoked AC activity revealed striking differences between response patterns of neurons from the core and belt areas. Most of core neurons displayed short latency, phasic responses, unlike the prevalence oflonger latency tonic responses in the belt area. It has been hypothesized that incoming signals could be processed differently by neurons in these areas. In thethesis we have investigated the intrinsic electrical properties of layer V pyramidal neurons by patch-clamp recording in acute rat AC slices. Results showed that the pyramidal neurons from the core AC are more excitable at rest...
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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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The study of functional and molecular properties of NMDA receptors
Vyklický, Vojtěch ; Vyklický, Ladislav (advisor) ; Chvátal, Alexandr (referee) ; Tureček, Rostislav (referee)
N-methyl-D-aspartate (NMDA) receptors are a subtype of ionotropic glutamate receptors, which mediate fast synaptic transmission in the vertebrate central nervous system and play a key role in synaptic plasticity. NMDA receptor overexcitation leads to cell death that underlies many serious neurological and psychiatric disorders. The aim of our research was to characterize novel drugs that modulate NMDAR activity. We have tested a series of newly synthesized steroidal compounds for their activity on recombinant receptors. We have demonstrated a relationship between the structure of the analogues of a naturally occurring neurosteroid pregnanolone sulfate and their ability to modulate NMDA receptors. The results of our experiments characterize the role of substituents at the steroid A and D ring. We have found specific substituents on carbons C3 and C17 of the steroidal skeleton that lead to a substantial increase of steroid efficacy and the ability to positively affect the ratio between the inhibition of stationary and non-stationary receptor activation. These two states represent receptors long-term activated (tonically) and synaptically activated (phasically), respectively. Using electrophysiological and optical methods in combination with mathematical modeling we have shown that the plasma membrane plays an...
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The role of neuroglia in the development of drug dependence
Stratilová, Mária ; Novotný, Jiří (advisor) ; Chvátal, Alexandr (referee)
Drugs of abuse, such as opioids and amphetamines, represents nowadays a serious global problem which has economical, psychological, social and medical impact. Investigation of drug effects on the nervous system has been a subject of many studies during the last few decades. The interest in neurons regarding this issues is rather constant, but currently there is increasing number of studies focused on neuroglia and their role in drug addiction. Many studies demonstrated that glia are not only a part of neuropile and do not have only supporting function, but they play an important role in communication between neurons, participate in modulation of neurotransmission and could produce factors such as cytokines and chemokines. However, there is a not much information about the effect of drugs of abuse on neuroglia and the presumed role of these cells in the development of addiction in not quite clear. This review aims to provide a brief survey of current knowledge on this topic.
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