|
|
Characterization of immune cells and monitoring changes of inflammatory proteins in minipig model of Huntington's disease
Butalová, Nikola ; Motlík, Jan (advisor) ; Janda, Jozef (referee)
The Huntington disease (HD) is a hereditary neuro-degenerative disorder caused by a mutation of the huntigtin gene that codes a protein of the same name. The mutated form of the huntigtin gene plays its part in many pathological interactions and influences a number of cellular mechanisms, including the immune system that could serve as a modifier of the neuropathology of the disease. The cells of the monocyte-macrophage system express cytokines whose production changes in relation to the activation of the cell. The presence of the mutated huntingtin protein in these cells renders them hyper-responsive to immunity incentives leading to changes in the production of cytokines. These differences are discernible a few years prior to the appearance of the symptoms. Therefore, the changes in the levels of certain cytokines could serve as appropriate biomarkers for monitoring of the onset of the disease and its progression. The HD pathogenesis includes an inflammation of the central neutral system. Inflammatory changes in peripheral tissues could reflect inflammatory processes in the central neural system. A miniature TgHD pig could represent an appropriate model organism for studying of the impact of the mHtt on the immune system. This model enables to observe a slow progression of the disease. Changes in...
|
|
|
Monitoring of the development of the Huntington's disease in transgenic minipigs with N-terminal part of human mutated huntingtin: biochemical and motoric changes of F0, F1 and F2 generation
Kučerová, Šárka ; Ellederová, Zdeňka (advisor) ; Klempíř, Jiří (referee)
Huntington's disease (HD) belongs to neurodegenerative disorders. It is a monogenic disease caused by trinucleotic CAG expansion in exon 1 of gene coding protein huntingtin. Even though the cause of HD is known since 1993, the pathophysiology and cure for HD reminds to be found. The animal models are being used for better understanding of HD. The most common animal models for HD are rodents, especially mice but it was also important to create large animal models, which will be more like human. Therefore, TgHD minipig was created in Academic of Science in Liběchov in 2009. This model was created by microinjection of lentiviral vector carrying N-terminal part of human HTT with 124 repetitive CAG in exon 1. This model is viable and in every generation, is part of the offspring transgenic. In this thesis, I specialized to biochemical and behavioral changes of this model. I compared transgenic and wild type siblings. I found that biochemical changes are manifested mostly by increased level of mtHtt fragments in testes and brain. In behavioral part of this thesis I established new methods for testing behavioral changes in this model. The introduced methods showed some changes between wild type and transgenic animals at the tested ages but these changes were not significant due to the low number of...
|
|
|
The role of mitochondria in the pathogenesis of Huntington's disease
Řeháková, Kateřina ; Hansíková, Hana (advisor) ; Kohoutová, Michaela (referee)
Huntington's disease is a neurodegenerative disease affecting the nervous system. It is caused by the mutation of the huntingtin protein coding gene. The mutation is manifested by the multiplication of CAG triplets. Huntingtin is present more in the cytoplasm. It interacts with many proteins and has roles also in transcription and cell transport. Huntingtin also participates in correct regulation of embryonic development and development of nerve tissue. Mutant huntingtin causes oxidative stress, mitochondrial biogenesis and OXPHOS disorders. Diagnosis of Huntington's disease is based on a laboratory examination of the presence of an allele predisposing to the disease. Clinical imaging is also an important part of diagnostics. The patient exhibits uncontrollable choreatic body movements and dementia. The aim of the thesis was to describe the main characteristics of Huntington's disease with the focus of HD on mitochondrial energy metabolism. In the practical part, the aim was to analyze mitochondrial respiration by high resolution polarography in a set of 3 fibroblast lines from patients with Huntington's disease. Polarographic analyses showed that respiration of HD fibroblasts was decreased in comparison with controls. We found that complex II of oxidative phosphorylation was most affected in HD...
|
|
|
Protein analysis of selected mitochondrial proteins in the muscle tissue of porcine model of Huntington's disease
Dosoudilová, Žaneta ; Klempíř, Jiří (advisor) ; Baxa, Monika (referee)
Huntington's disease (HD) is an autosomal dominant hereditary neurodegenerative disease characterized by motor, cognitive and behavioral disorders. HD is caused by expansion of CAG triplet (cytosine-adenosine-guanine) located in a gene on the short arm of the fourth chromosome. This expansion encodes an aberrant polyglutamine chain in the protein huntingtin. Physiological and mutated huntingtin (in case of HD) are expressed in almost all tissues and influences many cellular functions. The prevalence of HD in population is about 1 per 10.000. The disease is currently incurable and its mechanisms are not sufficiently understood. Besides affecting the central nervous system HD also affects peripheral tissues, including skeletal muscles. HD disrupts mitochondrial function and damages oxidative phosphorylation system, which has the task of producing energy in the form of ATP in cells. Research of transgenic minipig model for HD could help elucidate the mechanisms of disease's pathogenesis and potential therapeutic strategy. In this diploma thesis, immunodetection with help of specific antibodies to detect changes in amount of 14 selected mitochondrial proteins in skeletal muscle tissue of three age groups of transgenic HD minipigs - 24, 36 and 48 months old was used. Gradual progression in reduced...
|
|
|
Interplay between mutated huntingtin and precancerogenous status in mammalian cells.
Butalová, Nikola ; Baxa, Monika (advisor) ; Koudelková, Lenka (referee)
Huntington's disease is a serious hereditary disorder that causes mortification of neurons. The disease affects individuals around the age of 40. Its characteristics are involuntary movement of the limbs and a progressive dementia. This disorder is currently without any treatment and always ends with patient dying within a period of 15 years after the first symptoms are discovered. Special relation between Huntington's disease and malign neoplasia was observed at the end of the 20th century. This relation shows lower degree of cancer among the patients with this neurodegenerative disorder compared to the general population. An expanded sequence of a CAG section probably protects these persons against advancement of cancer. Creation of an applicable experimental model with characteristic highly resembling human body was necessary for superior research of this disease. This model can be represented by a transgene mini pig carrying a mutated protein huntingtin - tgHD pig. Another model of a mini-pig showing hereditary occurrence of malign lesions - MeLiM pig was created to research cancer. These two specific lines of mini-pigs were crossbred resulting in not only piglets with melanoma and transgene piglets with mtHTT but also transgene piglets having melanoma at the same time. Furthermore the same...
|
|
|
Modelovanie ochorenia a štúdium regeneračných procesov v Huntingtonovej chorobe a ALS in vivo
Hruška-Plocháň, Marián
Neurological disorders affect more than 14% of the population worldwide and together with traumatic brain and spinal cord injuries represent major health, public and economic burden of the society. Incidence of inherited and idiopathic neurodegenerative disorders and acute CNS injuries is growing globally while neuroscience society is being challenged by numerous unanswered questions. Therefore, research of the CNS disorders is essential. Since animal models of the CNS diseases and injuries represent the key step in the conversion of the basic research to the clinics, we focused our work on generation of new animal models and on their use in pre-clinical research. We generated and characterized transgenic minipig model of Huntington's disease (HD) which represents the only successful establishment of a transgenic model of HD in minipig which should be valuable for testing of long term safety of HD therapeutics. Next, we crossed the well characterized R6/2 mouse HD model with the gad mouse model which lacks the expression of UCHL1 which led to results that support the theory of "protective" role of mutant huntingtin aggregates and suggest that UCHL1 function(s) may be affected in HD disturbing certain branches of Ubiquitin Proteasome System. Traumatic spinal cord injury and Amyotrophic Lateral...
|
|
|
Changes in beta-catenin expression during ontogenesis in the transgenic minipigs for human mutant huntingtin
Žižková, Martina ; Motlík, Jan (advisor) ; Jarkovská, Karla (referee)
Huntington's disease (HD) is an inherited autosomal dominant neurodegenerative disorder caused by an unstable expansion of the CAG repeat sequence within the huntingtin gene. Huntingtin associates with ubiquitin-proteasome system that ensures degradation of particular proteins including β-catenin which is an important molecule whose equilibrated degradation is necessary for the proper functioning of the Wnt signaling pathway. The binding of β-catenin to the destruction complex is altered in HD, leading to the toxic stabilization of β-catenin. The main goal of my thesis was to determine whether the accumulation of β-catenin due to the presence of mutant huntingtin is also characteristic of Liběchov minipigs, a large animal model of Huntington's disease stably expressing N-truncated human mutant huntingtin. Using immunoblot and specific antibodies, we have revealed age-dependent accumulation of mutant huntingtin in transgenic minipigs. Unlike endogenous huntingtin, no decrease of the level of mutant huntingtin was observed in the striatum of transgenic animals. Surprisingly, this was followed by a decrease of phosphorylated β-catenin. Nevertheless, our results demostrate the accumulation of β-catenin in mesenchymal stem cells isolated from the oldest boars during ontogenesis. Furthermore, we have revealed a...
|
|
|
Huntington's disease modeling and stem cell therapy in spinal cord disorders and injury
Hruška-Plocháň, Marián ; Motlík, Jan (advisor) ; Bjarkam, Carsten (referee) ; Roth, Jan (referee)
Neurological disorders affect more than 14% of the population worldwide and together with traumatic brain and spinal cord injuries represent major health, public and economic burden of the society. Incidence of inherited and idiopathic neurodegenerative disorders and acute CNS injuries is growing globally while neuroscience society is being challenged by numerous unanswered questions. Therefore, research of the CNS disorders is essential. Since animal models of the CNS diseases and injuries represent the key step in the conversion of the basic research to the clinics, we focused our work on generation of new animal models and on their use in pre-clinical research. We generated and characterized transgenic minipig model of Huntington's disease (HD) which represents the only successful establishment of a transgenic model of HD in minipig which should be valuable for testing of long term safety of HD therapeutics. Next, we crossed the well characterized R6/2 mouse HD model with the gad mouse model which lacks the expression of UCHL1 which led to results that support the theory of "protective" role of mutant huntingtin aggregates and suggest that UCHL1 function(s) may be affected in HD disturbing certain branches of Ubiquitin Proteasome System. Traumatic spinal cord injury and Amyotrophic Lateral...
|
|
|
Biomedical models of Huntington disease
Žižková, Martina ; Motlík, Jan (advisor) ; Moravec, Jan (referee)
Huntington's diease is a dominant inherited neurodegenerative disorder that is caused by an expansion of a CAG repeats within a huntingtin gene. Mutant protein causes a neuron degeneration in a brain of HD pacients which leads to a motor abnormalities and personality decay. This disease is very malign because of its late onset. An equal therapy does not exist yet, but a lot of research teams focus on designig a suitable medical treatment. It is necessary to create animal models of Huntington disease which can be used for testing the therapies. In my work I aim to summarize the animal models of HD which are used in research. A rodent model is the most common due to its low price and easy breeding. However, more important are human related large animals like sheep, pigs or non-human primates. The principal criterion of animal model is its method of creation. We can divide the models into two categories, genetic and non-genetic. The memebers of the first one are able to reproduce better expression of human Huntington disease. Generation of animal models of HD leads to better comprehension the principles of HD, and developing an equal therapy for HD pacients.
|
guest ::
