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Role of NMDA NR1 subunit in pathophysiology of schizophrenia
Vrajová, Monika ; Horáček, Jiří (advisor) ; Blahoš, Jaroslav (referee) ; Vyklický, Ladislav (referee)
Our work is focused on the role of NR1 subunit of N-methyl-D-aspartate receptor in pathophysiology of schizophrenia. In animal model using separately or in combination, antisense oligodeoxynucleotide (aODN) for NR1, NR2A and NR2B subunit of NMDAR, we affected expression of these proteins in rat hippocampus. We assessed prepulse inhibition of acoustic startle reaction (PPI) in rats and protein expression of NMDAR subunits and expression of PSD proteins. There were significant differences in expression of PSD-95 and NR1 between groups. Application of aODN (NR2A, NR2B) was associated with a significant decrease of PSD-95. PPI and expression of NR2A, NR2B and PSD-93 were not changed after aODN application.The next part of the work concentrates on a human post mortem study. To assess actual changes in the expression of the NR1 subunit and its isoforms, we measured absolute differences in the levels of mRNA/protein for panNR1, as well as the individual mRNA/protein isoforms in the post mortem left/right hippocampus of patients with schizophrenia in comparison with non-psychiatric subjects. There were no significant differences in the panNR1 subunit mRNA expression, but the absolute left/right differences were much more pronounced in the patients with schizophrenia. The expression of splice variants in the...
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Neurosteroid modulation of ligand-gated ion-channel activity
Krausová, Barbora ; Vyklický, Ladislav (advisor) ; Moravec, Jan (referee)
The term neurosteroids refers to steroids that are synthetized in the nervous tissue from cholesterol or steroidal precursors from peripheral sources. These compounds affect the neuronal excitability by modulating the function of some ligand-gated ion channels. NMDA (N methyl D aspartate) receptors are glutamate gated ion channels involved in excitatory synaptic transmission, synaptic plasticity and excitotoxicity. GABAA ( aminobutyric acid type A) receptors mediate most of the inhibitory synaptic transmission in the mammalian brain and are targeted by many clinically important drugs. Function of NMDA and GABAA receptors can by affected by neurosteroids, both positively and negatively. The aim of this work is to summarize the current knowledge about the neurosteroid effects on the function of GABAA a NMDA receptors and suggest the physiological role and the potential therapeutic use of the neurosteroids as a regulator of some functions of the central nervous system.
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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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název v anglickém jazyce není uveden
Šámal, Filip ; Haninec, Pavel (advisor) ; Vyklický, Ladislav (referee) ; Zvěřina, Eduard (referee)
Traumatic injury of peripheral nerves represents an important area of present-day clinical and experimental interest. Despite a large quantity of experimental data acquired in an intense research of this topic carried out in recent years, traumatic affection of peripheral nerves continues to be a complex surgical problem. Injury of peripheral nerves often results in a permanent and serious impairment in the patients' lives connected with the loss of the affected peripheral nerve function. If an injured nerve is given a timely and correct treatment, these consequences can be prevented to a certain extent. In the current practice this implies the use of all available methods of peripheral nerves surgery, primarily microsurgical operation technique while observing some of the basic principles essential for a good functional reinervation. This mainly applies to tension-free suture. In some anatomic locations and despite observing the above-mentioned facts, it is still difficult to reach good functional results. This primarily applies to brachial plexus injury which is one of the most complicated and serious of all lesions of peripheral nerves. Out of the total number of nerve injuries they are represented in large sets by three to seven percent. In his set, Midha reports 1.2% of patients affected by the...
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The influence of expression and polymorphism of proinflammatory genes on kidney transplant outcome
Hřibová, Petra ; Vyklický, Ladislav (advisor) ; Jirsa, Milan (referee) ; Stříž, Ilja (referee)
Acute rejection and chronic transplant nephropathy are the main complications after kidney transplantation. A broad spectrum of cytokines, chemokines and growth factors are involved in their etiology. The aim of this submitted dissertation was to find the influence of polymorphisms in selected genes and their intrarenal expression on kidney graft outcome. All studies were designed to follow a large cohort of individuals in order to be able to elucidate some unclear and inconsistent published results. 1. Cytokines and chemokines TGF-β1, TNF-α, IL-6, IL-10, MCP-1 and RANTES are up-regulated in different rates in acute rejection, chronic transplant nephropathy and also in other causes of kidney graft dysfunction. 2. High intrarenal expression of TGF-1 and MCP-1 mRNA in CAN predicts a higher risk of kidney graft dysfunction in the long-term. Also, the kidney graft survival is significantly shorter. 3. Intrarenal gene expression profile of TGF-1, TNF-, IL-6, IL-10, MCP-1 and RANTES is different during various causes of graft dysfunction. The intrarenal expression level cannot be used for diagnostic purposes, but it can alert higher immunological activity in kidney graft, which can lead to earlier failure of renal functions. 4. We did not confirm an association of TNF--308G/A, MCP-1 -2518 A/G, RANTES-...
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A study of the modulation of recombinant and native NMDA receptors by physical and chemical factors
Cais, Ondřej ; Vyklický, Ladislav (advisor) ; Blahoš, Jaroslav (referee) ; Chvátal, Alexandr (referee) ; Šťastný, František (referee)
Excitatory synaptic transmission in mammalian CNS is mostly mediated by ionotropic glutamate receptors. NMDA receptors, one of three subclasses of this ligand-activated ion channels family, are involved in memory formation and learning and also play a role in the pathogenesis of various neurodegenerative diseases. Current knowledge about NMDA receptors function is predominantly based upon results of in vitro experiments conducted at room temperature, far from physiological. The aim of this PhD thesis was to describe the temperature dependence of NMDA receptors. We determined the rate constants that characterise each step in the mechanism of recombinant NR1/NR2B receptors activation in the temperature range 25-45řC. The receptor desensitization, resensitization and glutamate unbinding turned out to be the most temperature sensitive of these processes. In addition to that, we described the temperature dependence of deactivation kinetics in various experimental models of NMDA receptors (both recombinant and native). The second part of the thesis focused on the modulation of NMDA receptors function by steroid compounds derived from pregnanolone sulfate, an endogenously occuring neurosteroid. We tested 21 steroids that showed various degree of ability to inhibit (or, in one case, potentiate) the current...
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Metabotropic glutamate receptors: mechanism of activation
Hlaváčková, Veronika ; Blahoš, Jaroslav (advisor) ; Vyklický, Ladislav (referee) ; Martásek, Pavel (referee) ; Konvalinka, Jan (referee)
Any living organism receives constantly many signals that have to be evaluated and weighted to respond in an appropriate way. To perform all functions needed for precise control of homeostasis and for communication with the surrounding environment, signals coming from the outside are recognized and transferred into modulation of intracellular signaling cascades. These mediate response to the extracellular stimulus as well as intercellular communication. Cell communication is mediated by several types of receptors, located either intracellularly (including nuclear receptors) that modulate gene transcription and receptors localized on plasma membrane. Cell membrane receptors are transmembrane proteins that are divided into three superfamilies according to their structure and principles of signal transduction. These are ion channel-linked receptors, enzyme-linked receptors and G-protein-coupled receptors (GPCRs). GPCRs comprise the biggest family of membrane receptors and are one of the largest gene families in general. They are encoded by about 1% of genes in mammals. Many of them bind sensory ligands (rhodopsin, taste and olfactory receptors), but others also recognize ions, amino acids, nucleotides, peptides and large glycoproteins (1). They play a crucial role in such distant physiological functions as...
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