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Úloha Adenylát kinázy 1 v aktivaci a metabolismu imunitních buněk larev \kur{Drosophila melanogaster}
KAISLEROVÁ, Nikola
The aim of this thesis was to study the role of Adenylate kinase 1 (Ak1) in the immune system of Drosophila melanogaster larvae upon the infection by parasitoid wasp Leptopilina boulardi. Using the immune specific Ak1 RNA interference, it was analyzed the effect of Ak1 reduction on the immune response and viability of Drosophila. The importance of Ak1 was also evaluated within the metabolism of immune cells. It has been shown that Ak1 is crucial in energy metabolism of immune cells and important for the proper functioning of immune system.
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Vliv cytosolické nukleotidázy cN-IIIB na imunitní odpověď u \kur{Drosophila melanogaster}
DOLEJŠKOVÁ, Tereza
Adenosine is a key signal molecule of the metabolic switch, a crucial process of metabolic changes in Drosophila melanogaster upon parazitoid wasp infection. However, some components of the adenosine creation pathway have yet to be discovered. We studied a potential convertor of AMP to adenosine in immune response, cN-IIIB, a cytosolic nucleotidase known to accept the methylated RNA cap nucleotide 7-methylguanosine as a substrate and to protect cells against undesired incorporation of this nucleotide into nucleic acids. We suggest this trait can be potentially important in the metabolic switch.
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Role makrofágy produkovaného cytokinu IMPL2 v regulaci systemického metabolismu během bakteriální infekce u \kur{Drosophila melanogaster}
VARGOVÁ, Hana
The main goal of this bachelor thesis is to verify whether ImpL2, produced by activated immune cells, is responsible for changes in systemic metabolism during the immune response during the acute phase of bacterial infection in Drosophila melanogaster. The theoretical part includes previously known information about the immune system of mammals and D. melanogaster, especially about polarization and cellular metabolism of macrophages, as well as knowledge about the roles of the ImpL2 gene and its mammalian homologue Igfbp7. The practical part of the bachelor thesis deals with the role of the ImpL2 gene in the induction of changes in systemic metabolism during the immune response. The effect of the cytokine IMPL2 on selected metabolic genes in adipose tissue after Streptococcus pneumoniae infection was tested by means of macrophage-specific RNA interference. The work also clarifies whether this signaling factor has an effect on the concentration of selected metabolites in circulation and in macrophages during the acute phase of bacterial infection.
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Vytvoření a charakterizace transgenních linií drozofil manifestující neurodegenerativní onemocnění spinocerebelární ataxi typu 1 (SCA1)
JIČÍNSKÁ, Veronika
The aim of this thesis was to generate and characterize transgenic lines of Drosophila melanogaster manifesting spinocerebellar ataxia type 1 (SCA1), using GAL4/UAS system. Specific driver lines were used to express ataxin gene in pan neuronal, eye and motor neurons. Adult males from different crosses expressing abnormal ataxin gene with parallel controls aged 5 day and 30 day were used for all experiments. The effects of such expression on mobility, longevity and neurodegenerative changes in the brain was studied. Moreover, specific techniques such are RT-PCR, and western blotting were used to test expression of specific genes and to quantify level of specific proteins. Neurodegenerative changes in transgenic lines were documented using confocal microscopy. The expression of specific genes such as ataxin and matrix metalloproteinase 1 and 2 (dMMP1, dMMP2) was studied in all used fly lines. The results showed that gene expression of hAtXN1 was higher in lines manifesting SCA1 compared to controls. However, the trend of MMP expression was not completely clear and will be the subject of future research.
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Vliv paralytického jedu lumčíka, \kur{Habrobracon hebetor}, na svalovou tkáň zavíječe, \kur{Ephestia kuehniella} a octomilky, \kur{Drosophila melanogaster}
PODLAHOVÁ, Šárka
The ectoparasitoid wasp Habrobracon hebetor uses its effective venom for inducing neuromuscular paralysis to ensure the proper development of its progeny in the host attacked. However, the associated mechanism of the paralysis is not known yet. The action of the venom was studied in the muscle tissue of its host larvae Ephestia kuehniella and in the model organism Drosophila melanogaster using transmission electron microscopy (TEM). Preparation of the samples for TEM was optimized to protect tissue integrity and to minimalize artefacts. The results revealed that the wasp venom affected neither the organization of muscle fibres nor the structure of mitochondria. Furthermore, potential D. melanogaster mutants of glutamate receptor subunits GluRIIA and GluRIIB were analyzed, and the role of glutamate receptors in regulation of paralysis was discussed.
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Důležitost systemické inzulínové rezistence pro imunitní odpověď
STRYCH, Lukáš
The aim of this thesis was to study insulin resistance in larval peripheral tissues of Drosophila melanogaster upon the infection by parasitoid wasp Leptopilina boulardi. Firstly, the state of the insulin signalling during the immune response was analysed by measuring gene expression and phosphorylation of Akt. Subsequently, the importance of insulin resistance in peripheral tissues for the effectivity of the immune response was tested. The effect of Jak-Stat signalling on inducing insulin resistance in larval muscle was evaluated as well.
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Role of adipokinetic hormone during starvation in Drosophila
KUTHANOVÁ, Michaela
The role of adipokinetic hormone (Drome-AKH) in maintaining the levels of basic nutrients, under starvation conditions, was studied using Drosophila melanogaster mutants with AKH deficiency (Akh1) and AKH abundance (EE-Akh). Our results showed lipids as the main energy reserve in Drosophila, and their physiological level and metabolism were shown to be under the control of AKH. AKH abundance in the body resulted in lower levels of triacylglycerols and diacylglycerols than in the controls, probably due to a more intensive metabolism; interestingly, there was a disproportional representation of fatty acids in triacylglycerols and diacylglycerols in Drosophila. Lower level of glycogen and its partial control by AKH suggest its lesser role as the storage substance. However, maintenance of free carbohydrate level in Drosophila seemed to be critical; when glycogen stores are exhausted, carbohydrates are synthesized from other sources. Protein levels and their alterations, under starvation, did not seem controlled by AKH. AKH-deficient flies were more resistant while AKH-abundant flies were more sensitive to starvation; females were found to be more resistant than males, regardless of the AKH level, probably due to higher body mass and higher amount of nutrients. However, in accordance with the level of all nutrients, that of AKH also gradually decreased with prolonged starvation
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CRISPR/Cas9 gene editing in Drosophila melanogaster
KALTENBÖCK, Konstantin
The aim of this thesis was to predict functional sites in the timeless gene of Drosophila melanogaster, to design fitting gRNAs for the identified sites to create transformation vectors for fly transformations and to induce CRISPR/Cas9 mediated target mutations by crossing gRNA expressing flies with Cas9 expressing flies.
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Juvenile hormone role on metabolic switch during immune-response
TSCHERNUTH, Sebastian
The role of Juvenile hormone during immune response was investigated in Drosophila melanogaster larvae. The levels of Juvenile hormone were measured directly by mass spectrometry and indirectly by monitoring Krüppel Homologue expression. Neither of those approaches were successful. We genetically manipulated JH production and signalling, infected the larvae with parasitoid wasps and found a significant difference in immune strength between larvae with and without functional JH production.
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