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The role of anillin in the growth cone of neurons
Tomášová, Štěpánka ; Libusová, Lenka (advisor) ; Vinopal, Stanislav (referee)
During embryonal development, axons of newly differentiated neurons need to properly interconnect and create a functional neuronal network. To achieve this, the cell requires a growth cone. The growth cone is a highly dynamic structure at the end of growing axons that serves both as the navigator and the propeller. Crosstalk between actin and microtubules is vital for proper axonal pathfinding. But the exact mechanism of this cooperation remains unknown. This diploma thesis investigates the possible role of a candidate scaffolding protein called anillin in this process. Anillin has been studied in two human cell lines. SH-SY5Y neuroblastoma cell line was used for overexpression and siRNA knock-down experiments. Anillin overexpression led to perturbed neurite morphology and growth cone dynamics in SH-SY5Y cells, whereas cells with lower anillin expression had fewer neurites. Next, neurons differentiated from human iPSC (induced pluripotent stem cells) expressing endogenous fluorescently tagged anillin were studied. Local dynamic high concentration spots of anillin have been observed at the base of cell protrusions of differentiating neurons. These anillin flares appeared during cell migration, early neurite initiation, and in newly created growth cones. These results suggest that anillin plays a...
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MAP code and regulation of microtubule-based processes
Karhanová, Adéla ; Lánský, Zdeněk (advisor) ; Tomášová, Štěpánka (referee)
Microtubule associated proteins (MAPs) are considered as key regulators of molecular trafficking in cells. Even though their malfunctioning results in severe pathologies, such as neurodegenerative disorders, the regulatory roles of these proteins remain under debate. Since MAPs bind to the cytoskeleton, this structure has to be vital for the function of MAPs. Microtubules, a highly dynamic type of cytoskeletal structure, have been given extra attention due to their association with cell division and vital functions in neurons. Microtubules can undergo post-translational modifications that affect molecular motors as well as binding of other proteins, such as MAPs. Whether post-translational modifications of microtubules regulate the distribution of MAPs is so far not sufficiently documented. However, MAPs have been shown to cooperatively form cohesive envelopes on the microtubules and thereby regulate the access of motors and severing enzymes. As there are many types of MAPs and they are mutually exclusive, a hypothesis of a regulatory 'MAP code' emerged recently in the literature. Using available literature, this review will try to introduce the new model of MAP code and provide some background information on previous research on this topic.
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The role of anillin in the growth cone of neurons
Tomášová, Štěpánka ; Libusová, Lenka (advisor) ; Vinopal, Stanislav (referee)
During embryonal development, axons of newly differentiated neurons need to properly interconnect and create a functional neuronal network. To achieve this, the cell requires a growth cone. The growth cone is a highly dynamic structure at the end of growing axons that serves both as the navigator and the propeller. Crosstalk between actin and microtubules is vital for proper axonal pathfinding. But the exact mechanism of this cooperation remains unknown. This diploma thesis investigates the possible role of a candidate scaffolding protein called anillin in this process. Anillin has been studied in two human cell lines. SH-SY5Y neuroblastoma cell line was used for overexpression and siRNA knock-down experiments. Anillin overexpression led to perturbed neurite morphology and growth cone dynamics in SH-SY5Y cells, whereas cells with lower anillin expression had fewer neurites. Next, neurons differentiated from human iPSC (induced pluripotent stem cells) expressing endogenous fluorescently tagged anillin were studied. Local dynamic high concentration spots of anillin have been observed at the base of cell protrusions of differentiating neurons. These anillin flares appeared during cell migration, early neurite initiation, and in newly created growth cones. These results suggest that anillin plays a...
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Prokaryotic tubulin homologs
Tomášová, Štěpánka ; Libusová, Lenka (advisor) ; Schwarzerová, Kateřina (referee)
Cytoskeletal proteins form very important structures in eukaryotic cells. It was demonstrated in last years that cytoskeletal proteins are also an important part of prokaryotic cells. One of the cytoskeletal families is the tubulin family. The tubulins are known to form dynamic microtubules which are important for cell division and other processes in eukaryotic cells. Proteins homologous to tubulin were discovered in Prokaryotes, based on their similarity to the tubulin GTP-binding region. This thesis describes the structure and function of known tubulin homologs from bacteria and archaea and compares them to their eukaryotic counterplayers.
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