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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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Studies of polyomavirus trafficking from late endosomes towards the cell nucleus
Štach, Martin ; Forstová, Jitka (advisor) ; Němečková, Šárka (referee)
Mouse polyomavirus (MPyV) is a model virus of the Polyomaviridae family. Polyomaviruses are small non-enveloped DNA viruses. They cause severe problems to immunocompromised patients. Their oncogenic potential is known in animals and humans. Trafficking of MPyV within the cell is not clear yet. The virus enters via smooth monopinocytic vesicles and continues to early and late endosomes. From there, the virus is transported to the ER by unknown mechanism. It bypasses Golgi aparatus (GA). One possible pathway is from late endosomes to trans-Golgi network (TGN) facilitated by Rab9 GTPase and then in COPI vesicles to the ER. In this thesis, the effect of inhibitors of retrograde transport (Brefeldin A, Golgicide A) on MPyV infection was evaluated. Brefeldin A is not completely specific; it has effect on whole endosomal system. Golgicide A causes specific disruption of transport via TGN and GA. Both inhibitors suppressed infection of MPyV. Confocal microscopy revealed colocalization of some MPyV virions with markers of TGN and COPI vesicles. MPyV didn't colocalize with cis-Golgi marker. Unfortunately, the effect of overexpression of Rab9 dominant negative mutant couldn't been evaluated due to its high cytotoxicity. However, overexpression of wild type Rab9 slightly increased infectivity. The results...
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Studies of polyomavirus trafficking from late endosomes towards the cell nucleus
Štach, Martin ; Forstová, Jitka (advisor) ; Němečková, Šárka (referee)
Mouse polyomavirus (MPyV) is a model virus of the Polyomaviridae family. Polyomaviruses are small non-enveloped DNA viruses. They cause severe problems to immunocompromised patients. Their oncogenic potential is known in animals and humans. Trafficking of MPyV within the cell is not clear yet. The virus enters via smooth monopinocytic vesicles and continues to early and late endosomes. From there, the virus is transported to the ER by unknown mechanism. It bypasses Golgi aparatus (GA). One possible pathway is from late endosomes to trans-Golgi network (TGN) facilitated by Rab9 GTPase and then in COPI vesicles to the ER. In this thesis, the effect of inhibitors of retrograde transport (Brefeldin A, Golgicide A) on MPyV infection was evaluated. Brefeldin A is not completely specific; it has effect on whole endosomal system. Golgicide A causes specific disruption of transport via TGN and GA. Both inhibitors suppressed infection of MPyV. Confocal microscopy revealed colocalization of some MPyV virions with markers of TGN and COPI vesicles. MPyV didn't colocalize with cis-Golgi marker. Unfortunately, the effect of overexpression of Rab9 dominant negative mutant couldn't been evaluated due to its high cytotoxicity. However, overexpression of wild type Rab9 slightly increased infectivity. The results...
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|
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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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