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Distribution of mitochondrial uncoupling proteins in selected tissues from mice and rat
Alán, Lukáš ; Flachs, Pavel (referee) ; Ježek, Petr (advisor)
Mitochondrial uncoupling proteins (UCPs) belong to the superfamily of mitochondrial anion-carriers. The longest known is UCP1, predominantly expressed in brown adipose tissue, where it takes part in nonshivering thermogenesis. In the late 1990s were discovered other sequence homologs of UCP1 with tissue specific distribution. The Function of these "new" uncoupling proteins is still uncertain. It is assumed that each of the isoforms has a specific function depending on the type of tissue. This thesis showed differences in tissue transcription pattern between rat and mice using RT-PCR absolute quantification. Significant differences in pattern were found in lungs, brain and muscle. In each case UCP expression was higher in mice tissues. Mice lungs express mainly UCP2. The difference in mice brain is caused by ucp4 and ucp5 genes transcription and finally in muscle is highest content of UCP3 mRNA. We investigated whether any of ucp transcript can complement ucp2 transcripton in spleen or lungs of ucp2 -/- mice. We did not find any difference which can explain, that in isolated lung mitochondria of fasted ucp2-/- mice were uncoupled in state 4. In the last project, we found relationship between ucp2 transcription in insulinoma INS-1E cells and oxygen levels of the cultivation atmosphere.
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Steatosis of liver and mitochondrial dysfunction
Páleníčková, Eliška ; Balková, Patricie (referee) ; Novák, František (advisor)
Aim: To determine the effect of diet-induced steatosis in the development of mitochondrial dysfunction in the liver and hepatic sensitivity to the partial ischemia. Methods: Male Wistar rats (361 ± 8.8 g) were fed standard (SD) or high-fat diet (HFD). Partial ischemia was induced by short-term clamp (20 min) of vein porta two days before the end of the experiment. Results: Ten-week HFD administration lead due to increased ketogenesis the altered glucose tolerance elevated serum NEFA. We demonstrated the inhibitory effect of HFD on the respiratory capacity of mitochondria in vitro. HFD negatively affected the activity of antioxidant systems and stimulated the formation of lipoperoxides. Partial ischemia had no efect on the mitochondrial oxidative capacity but significantly elevated the oxidative stress. Conclusion: HFD administration lead to the development of fatty liver that was still not accompanied by biochemical markers of liver injury. Nevertheless, we proved the impairment of to mitochondrial respiratory capacity, signs of structural damage of mitochondria and the increased sensitivity to oxidative damage of the liver. Subject words: biochemistry, physiology Keywords: mitochondria, HFD, ischemia, respiratory chain, antioxidant systém, ROS
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Evolution of eukaryotic ABC transporters
Žárský, Vojtěch ; Doležal, Pavel (referee) ; Tachezy, Jan (advisor)
In the past twenty years there has been a lot of research done on ABC transporters. This group of membrane transporters was recognized as highly important due to its ubiquity in living organisms and due to the involvement of some of the ABC transporters in multidrug resistance of cancer cells and pathogens against chemotherapeutics. This medical aspect of ABC transporters was naturally the most important one for the majority of researchers. On the other hand, the biological aspects and evolution of many ABC transporters remained untouched. In this work I give an overview of ABC transporters of parasitic protozoa and focus on the evolutionary aspect of eukaryotic transporters and on mitochondrial ABC transporters, which are the most conserved ones among eukaryotic ABC transporters.
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Ferrous nanoparts localization in inner organelles
Solař, Jan ; Janoušek, Oto (referee) ; Čmiel, Vratislav (advisor)
This bachelor‘s thesis describes a behavior of iron nanoparticles in human mesenchymal stem cells. First section deals with methods of fluorescent labelling of organelles and nanoparticles and settings of confocal microscope for their detection. Next section describes the iron nanoparticles metabolism and accumulation in cells. Finally, there is a section about the developement of the software utility for the localization and for the quantitative analysis of the fluorescence organelles and nanoparticles inside the living cells.
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Similarity/dissimilarity analysis of genomic data on the basis of graphical representation
Těthal, Jiří ; Provazník, Ivo (referee) ; Maděránková, Denisa (advisor)
The work deals with the identification of species of animal through the density of nucleotids of mitochondrial gene CO1. In the first part the theory summarized information about DNA barcoding and the mitochondria, cellular organel that with this method is closely related. The second part deals with virtually comparing different sequences using the density of nucleotides. For this it was created program, which uses two functions, the first of the specified nucleotide sequence calculates the density and one can compare the density by distance methods.
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Comparison of mitochondrial DNA for species identification
Labounek, René ; Provazník, Ivo (referee) ; Maděránková, Denisa (advisor)
The work deals with the method of recognizing species on the analysis of mitochondrial DNA segment. This analysis and classification using segment gene called CO1 in literatures such as barcode of life. In the beginning of work is analyzed the mitochondrial theory of heredity and conditions of formation of barcode. Practical use is based on this theory in creating database of barcodes generated to different animal species. Data used for creating the library are drawn from public databases NCBI and BOLD Systems. The next part of this work concerns about methods of comparison of the individual barcodes to the others and especially to the barcode of human. Three main computing methods were used tore these analyses: Needleman-Wunsch algorithm, Smith- Waterman algorithm and comparison of similarities using distance matrix. This work also concerns about transformation of DNA molecule sequences from symbols to numeric formats, which is required for the distance matrix comparison method. Algorithms for searching for a barcode of a species and vice versa were created to ease the work with data.
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Genetic diversity of chosen horse breeds
JANKOVSKÁ, Tereza
The objective of the thesis is to clarify the findings about genetical biodiversity of horses, evaluation of the diversity and description of current state of genetic resources preservation. Firstly, the history is described, domestication of horses, breeds and biodiversity. From the genetic point of view, the thesis contains description of the genetical diversity, microsatellites, markers and mitochondria. There are described genetic sources and their preservation. Current quantities and thein progress are also mentioned.
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Assembly and annotation of a mitochondrial genome of kinetoplastid protist \kur{Perkinsela}.
DAVID, Vojtěch
This thesis is based on a collaborative project between Laboratory of Molecular Biology of Protists led by Prof. Julius Lukeš and the laboratory of John Archibald (Department of Biochemistry & Molecular Biology, Dalhousie University, Canada). My contribution to the project, as the name of the thesis suggests is to process DNA and RNA sequencing data from Neoparamoeba pemaquidensis isolate that correspond to the mitochondrion of Perkinsela, also called Ichthyobodo-related organism (IRO). The main aim is to characterize mitochondrial gene content and to attempt to understand the mechanism of U-insertion/deletion RNA editing in Perkinsela and its evolution.
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