|
|
Stereological quantification of striatal NPY-positive interneurons and their genetic modification by the AAV5 viral vector
Smolík, Matyáš ; Janíčková, Helena (advisor) ; Chvojková, Markéta (referee)
Different types of interneurons in the striatum are still relatively poorly understood, especially compared to other parts of the brain such as the cortex or hippocampus. One of the lesser-known types of striatal interneurons that has received increasing attention recently, are neuropeptide Y-expressing interneurons (NPY+ interneurons). Available work indicates that these interneurons are controlled by striatal cholinergic interneurons and thus they may serve as mediators of cholinergic modulation in the striatum. The first aim of this thesis is to determine the absolute number of NPY+ interneurons in the mouse dorsal striatum by stereology, using a transgenic mouse model that shows GFP expression in NPY+ neurons. The second aim of this thesis is to use stereology to determine the volume of the dorsal striatum that we are able to target with viral expression when using a standard stereotactic injection protocol commonly used in our laboratory. The third aim of this study is to test the behavioural effect of knock down of the beta2 subunit of the nicotinic acetylcholine receptor expressed by NPY+ interneurons in the dorsal striatum. The work should serve as a first step in a study that will ultimately allow us to develop a quantitative idea of the absolute and relative number of NPY+ interneurons in...
|
|
|
Angiogenesis in cerebral ischemia border zone
Nulíčková, Radka ; Otáhal, Jakub (advisor) ; Brime, Tufikameni (referee)
Title of work: Angiogenesis in the border zones of cerebral ischemia The aims of work: The aim of this diploma thesis was to determinate the parameter of the angiogenesis after focal brain ischemia induced by photothrombosis in P7 rat pups. This thesis should answer the question, if there is some difference in vascular density according to the distance from the lesion and if there is some diference between the vascular density in normal brain and the brain with the lesion. Materials and Methods: Cerebral ischemia was induced in P7 rat pups with photothombic method. Intravenous administration of bengal rose dye was followed by illumination of beam of green laser over the cortex for 5 minutes at 0,5s on/off cycles. Bengal rose dye is potent photosensitive substance and the ilumination resulted in agregation of trombocytes and thrombosis. Cerebral ischemia evolved due to the thrombosis. After two months, the animals were overdosed with urethane and transcardially perfused. Then thein brain were cryoprotected and then cut into 50µm slices and prepaired for histological staining. The stained slices were the investigated under mikroskope. The microvessels in the cortex were counted and the density was estimated by stereological method. Results: Two brain with ischemic lesion and two control brain were...
|
|
|
Anatomical study of shade and sun European beech leaf under elevated CO2.
Horská, Jana ; Lhotáková, Zuzana (advisor) ; Urban, Otmar (referee)
The present Master thesis focuses on evalution of CO2 concentration and irradiance on selected leaf anatomical parameters of European Beech (Common Beech) Fagus sylvatica L.. The process of photosynthesis is remarkably determined by numerous extrenal factors, among them by atmoshperic CO2 concentration and irradiance and is closely correlated with leaf anatomical parameters. One of these most important anatomical parameters affecting the net assimilation rate is an internal leaf surface, which corresponds to mesophyll area avialable for gas exhcange. Experimental material of the study was sampled from the leaves of juvenile trees of F. sylvatica planted in 2005 and growing under ambient (390 ppm, AC) and elevated (700 ppm, EC) CO2 concentrations on the experimental site of the Global Change Research Center AS CR at Bílý Kříž in the Beskydy Mountains. Sun and shade leaves were sampled from trees of both CO2 treatments in two seasons 3 years apart (2009 and 2012). To determine leaf anatomical parameters, the stereological methods were applied, which yield unbiased estimation of measured parameters, particularly the Fakir method for internal leaf surface determination. The EC effect was observed on the leaves sampled in 2009 only in the decrease of proportion of intercellular spaces in mesophyll. In...
|
|
|
The Role of Lipids and Lipid Metabolizing Enzymes in Plant Autophagy
Krupař, Pavel ; Martinec, Jan (advisor) ; Hála, Michal (referee)
Plant autophagy is a crucial evolutionary conserved process for recycling cytoplasmic material under stress conditions or during development. The autophagic pathway is negatively regulated by TOR kinase, a versatile molecule modulating a wide range of cellular processes. In mammals, TOR kinase may be activated by phosphatidic acid, a vital signalling lipid. This thesis aims to prove the possible involvement of phospholipids in plant autophagy. I analysed the rate of primary root inhibition in knock-out mutants coding phospholipases in A. thaliana with induced autophagy, measured activity of lipid metabolising enzymes in wild type and atg10 mutant and observed autophagosome formation in selected mutants. Autophagosomes were labelled by fluorescent protein in vivo and by indirect immunolabelling in fixed samples. Using advanced stereological approach, I optimized a method for obtaining an unbiased estimate of autophagosome number in plant root cells.
|
|
|
Improved Methods of Image Acquisition and Analysis of Tissues and Cells by Confocal and Multi-Photon Microscopy
Chernyavskiy, Oleksandr ; Kubínová, Lucie (advisor) ; Hašek, Jiří (referee) ; Malínský, Jan (referee)
Univerzita Karlova v Praze Přírodovědecká fakulta Studijní program: Vývojová biologie (P1520) Studijní obor: Vývojová biologie (1501V000) Oleksandr Chernyavskiy Zdokonalené metody pro snímání obrazových dat a analýzu tkání a buněk pomocí konfokální a multifotonové mikroskopie Improved Methods of Image Acquisition and Analysis of Tissues and Cells by Confocal and Multi-Photon Microscopy Abstrakt disertační práce Školitel: RNDr. Lucie Kubínová CSc Praha, 2015 Abstract The aim of this study was to develop methods and approaches for image acquisition with subsequent image analysis of data, obtained by confocal and two- photon excitation microscopy as well as their combination, enabling new possibilities of visualization and assessment of information on biological tissues and cell structures in 3D and their measurement. We focused on methods that exploited advantages of confocal and multi-photon excitation microscopy. Our further aim was to demonstrate the applicability of non-invasive approach for in vivo applications, usefulness and the relevance of these methods in several special biological applications with emphasis on improved image acquisition, analysis and evaluation of real biological specimens. The present work was not oriented on just one specific biological problem, but rather to methodological...
|
|
|
Anatomical study of shade and sun European beech leaf under elevated CO2.
Horská, Jana ; Lhotáková, Zuzana (advisor) ; Urban, Otmar (referee)
The present Master thesis focuses on evalution of CO2 concentration and irradiance on selected leaf anatomical parameters of European Beech (Common Beech) Fagus sylvatica L.. The process of photosynthesis is remarkably determined by numerous extrenal factors, among them by atmoshperic CO2 concentration and irradiance and is closely correlated with leaf anatomical parameters. One of these most important anatomical parameters affecting the net assimilation rate is an internal leaf surface, which corresponds to mesophyll area avialable for gas exhcange. Experimental material of the study was sampled from the leaves of juvenile trees of F. sylvatica planted in 2005 and growing under ambient (390 ppm, AC) and elevated (700 ppm, EC) CO2 concentrations on the experimental site of the Global Change Research Center AS CR at Bílý Kříž in the Beskydy Mountains. Sun and shade leaves were sampled from trees of both CO2 treatments in two seasons 3 years apart (2009 and 2012). To determine leaf anatomical parameters, the stereological methods were applied, which yield unbiased estimation of measured parameters, particularly the Fakir method for internal leaf surface determination. The EC effect was observed on the leaves sampled in 2009 only in the decrease of proportion of intercellular spaces in mesophyll. In...
|
|
|
Will an increase in body temperature during status epilepticus in rat pups affect the extent and nature of damage to the hippocampus?
Chott, Robert ; Kubová, Hana (advisor) ; Otáhal, Jakub (referee)
Febrile seizures are epileptic seizures, arising in connection with febrile conditions in children of prechool age. In adults with epilepsy is often present a history of febrile status epilepticus, seizure whose duration is longer than 20 minutes. To study the role of febrile status epilepticus (FSE) in the development of epilepsy and neuronal damage, it is necessary to have a relevant animal models. This work is focused on the morphological analysis of the new created model of febrile status epilepticus, using a combination of short-term hyperthermia and chemical induced status epilepticus at 10 days old rats. In adulthood, the animals were examined by video/EEG monitoring, and then morphometric analysis. The aim of this study was to determine the importance of short-term hyperthermia during SE for neuropathological changes using stereological measurements of hippocampal volume.
|
|
|
Angiogenesis in cerebral ischemia border zone
Nulíčková, Radka ; Otáhal, Jakub (advisor) ; Brime, Tufikameni (referee)
Title of work: Angiogenesis in the border zones of cerebral ischemia The aims of work: The aim of this diploma thesis was to determinate the parameter of the angiogenesis after focal brain ischemia induced by photothrombosis in P7 rat pups. This thesis should answer the question, if there is some difference in vascular density according to the distance from the lesion and if there is some diference between the vascular density in normal brain and the brain with the lesion. Materials and Methods: Cerebral ischemia was induced in P7 rat pups with photothombic method. Intravenous administration of bengal rose dye was followed by illumination of beam of green laser over the cortex for 5 minutes at 0,5s on/off cycles. Bengal rose dye is potent photosensitive substance and the ilumination resulted in agregation of trombocytes and thrombosis. Cerebral ischemia evolved due to the thrombosis. After two months, the animals were overdosed with urethane and transcardially perfused. Then thein brain were cryoprotected and then cut into 50µm slices and prepaired for histological staining. The stained slices were the investigated under mikroskope. The microvessels in the cortex were counted and the density was estimated by stereological method. Results: Two brain with ischemic lesion and two control brain were...
|
|
| |
|
| |
guest ::
