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Role of β-arrestin in μ-opioid and TRPV1 receptor signalling
Nagy Marková, Vendula ; Novotný, Jiří (advisor) ; Blahoš, Jaroslav (referee) ; Roubalová, Lenka (referee)
β-Arrestin belongs to the protein family which has a huge impact not only on GPCR signaling, but its role exceeds the function of the membrane channel, its own signaling cascade, or as a scaffold protein, etc. Here we aimed to study β-arrestin roles on the MOR behaviour in the plasma membrane or -opioid receptor (MOR) signaling and effect on adenylyl cyclase (AC) function using the siRNA to decrease the expression of β-arrestin isoforms. Furthermore, we focused on investigating the role of β-arrestin on the crosstalk between MOR and TRPV1 channels, which are important parts of pain transduction. For this purpose, we used HEK293 cells that stably expressed MOR-YFP or transiently transfected with TRPV1-CFP. We observed that both β-arrestin isoforms have an effect on the lateral mobility of MOR in the plasma membrane and the silencing of one or another β-arrestin isoforms abolishes the effect of MOR agonists to affect its diffusion in the plasma membrane. Interestingly, silencing of β-arrestin1 diminish the internalization of MOR induced by the endogenous agonist endomorphin-2. On the other hand, silencing of β-arrestin2 did not abolish the endomorphin-2 induced MOR internalization. Moreover, both isoforms exhibit a distinct impact on the inhibition of AC induced by the agonists of MOR....
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Procedure for Granting International Protection in the Form of Asylum in the Czech Republic
Marková, Vendula ; Pítrová, Lenka (advisor) ; Petrmichl, Václav (referee)
Procedure for Granting International Protection in the Form of Asylum in the Czech Republic Abstract The content of this work is mainly the course of the asylum procedure from the submission of application for international protection to possible appeals, but also the definition of fundamental concepts, the historical development of the asylum institution, international, EU and national sources related to this procedure. In the introduction of this work I introduced and explained the different concepts without which it would be difficult to understand the asylum issue. In addition to outlining the historical development in this area, I have also taken the liberty of mentioning several major migration waves. Another important part of this thesis are the individual legal sources of this issue. The key documents of international regulation are described. Within the framework of Community law, the thesis lists the essential directives and regulations with a view to the necessary Europeanisation and subsequent incorporation into our legal system. The present work focuses on Europeanisation and the related influence of the CJEU case law on the interpretation of legal norms in the individual Member States of the European Union. It aims primarily to analyze and understand the various stages of the asylum procedure....
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Current approaches to cystic fibrosis therapy
Chmelíková, Barbora ; Kubíčková, Božena (advisor) ; Marková, Vendula (referee)
This bachelor thesis is focused on possible herapeutic methods for cystic fibrosis with main focus on modulators. The individual modulators are described in terms of their function, their differences and application possibilities. The thesis are also discuss other methods of treatment of cystic fibrosis, both for the respiratory tract and gastrointestinal tract. The thesis itself will be divided into three complementary parts. The first theoretical part defines the basic concepts of terms related to cystic fibrosis, the second follow-up practical part is focused on the comparison of the various methods of treatment of cystic fibrosis and the third part in the form of detailed analysis of the individual modulators. As for the methodology of the work, a method of literature search using secondary verified sources will be used. Keywords: cysticfibrosis, gene therapy, therapy, modulator, potentiator, correctors
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A study of molecular interactions of the μ-opioid receptor: the effect of biased ligands
Marková, Vendula ; Novotný, Jiří (advisor) ; Rudajev, Vladimír (referee)
G protein-coupled receptors (GPCRs) are the largest group of membrane-bound receptors. Transmission of signals into the cell interior is mediated through the interactions of these receptors with other signaling molecules. Nowadays, a great attention is devoted to biased ligands which are able to alter the conformation of the receptor in a specific way and thus distinctly affect its function. This diploma thesis was focused on a study of µ-opioid receptor (MOR), which is important in nociception. The aim of this study was to find out, how the activation of MOR by specific biased ligands (morphine, endomorphin-2 and DAMGO) affects the function and the interactions of MOR with potential molecular partners (for example G proteins or β-arrestin) A method of siRNA interference was used to knock down the following selected signaling molecules: Gαi1, Gαi2, Gαi3, Gαz and β-arrestin2. The effect of biased ligands on lateral mobility of MOR in the plasma membrane and on activity of adenylyl cyclase (AC) was examined under these conditions. We observed a possible involvement of Gαz subunit in the lateral mobility of MOR after the effect of morphine and endomorphin-2. The lateral mobility of MOR was significantly increased in cells lacking Gαi2 or Gαi3 or β-arrestin2. In this case the MOR was in inactive state....
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The role of proteostatic mechanisms in neurodegenerative diseases
Zezulová, Kristýna ; Vodička, Petr (advisor) ; Marková, Vendula (referee)
Protein homeostasis (proteostasis) plays an important role in maintaining normal cell function and viability. Neurons are particularly vulnerable to proteostasis dysregulation, resulting in damage, dysfunction, and neuronal death, as manifested in many neurodegenerative diseases. One of them is Huntington disease, hereditary neurodegeneration with autosomal dominant inheritance. Expansion of the CAG repeats in the huntingtin gene is translated into an abnormally long glutamine chain in huntingtin protein, leading to disruption of neuronal proteostasis. The primary affected area of the brain is the striatum of the basal ganglia. Disease is progressive, the onset of symptoms usually occurs in adulthood, and after many years leads to the death of the patient. Despite intensive research, disease pathology is still not fully understood, treatment is still only symptomatic and new studies, together with a deeper understanding, also raise many new questions. Through the complexity of the issue, the study of proteostasis in neurodegeneration can bring not only possible implications for therapy, but also could go deeper into the understanding of stress, memory or aging processes.
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A study of molecular interactions of the μ-opioid receptor: the effect of biased ligands
Marková, Vendula ; Novotný, Jiří (advisor) ; Rudajev, Vladimír (referee)
G protein-coupled receptors (GPCRs) are the largest group of membrane-bound receptors. Transmission of signals into the cell interior is mediated through the interactions of these receptors with other signaling molecules. Nowadays, a great attention is devoted to biased ligands which are able to alter the conformation of the receptor in a specific way and thus distinctly affect its function. This diploma thesis was focused on a study of µ-opioid receptor (MOR), which is important in nociception. The aim of this study was to find out, how the activation of MOR by specific biased ligands (morphine, endomorphin-2 and DAMGO) affects the function and the interactions of MOR with potential molecular partners (for example G proteins or β-arrestin) A method of siRNA interference was used to knock down the following selected signaling molecules: Gαi1, Gαi2, Gαi3, Gαz and β-arrestin2. The effect of biased ligands on lateral mobility of MOR in the plasma membrane and on activity of adenylyl cyclase (AC) was examined under these conditions. We observed a possible involvement of Gαz subunit in the lateral mobility of MOR after the effect of morphine and endomorphin-2. The lateral mobility of MOR was significantly increased in cells lacking Gαi2 or Gαi3 or β-arrestin2. In this case the MOR was in inactive state....
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β-Arrestin and its role in signal transduction
Marková, Vendula ; Novotný, Jiří (advisor) ; Nerandžič, Vladimír (referee)
β-Arrestin is a ubiquitous protein in cells, where it is involved in signal transduction and can affect different cellular processes. β-Arrestin cooperates with G protein-coupled receptors (GPCRs). Binding of β-arrestin to a receptor after its activation by a relevant ligand results in attenuation of signal transduction through the cognate G proteins, the process called desensitization, which can be associated with receptor intrenalization. Besides that, β-arrestin acts as adaptor for different molecules, which participate in signal transduction. β-Arrestin also has a role in a regulation of transcription in the cell nucleus. Finally, β-arrestin is explored in research focused on the development of a new type of drugs, so called biased ligands. After binding to a GPCR, these ligands can initiated only one specific activity of the receptor and affect relevant signaling cascades. Powered by TCPDF (www.tcpdf.org)
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