Annotation: Human cells of neuronal origin represent an excellent tool for the investigation of neuropathogenesis of TBE. The maturation, replication process of tick-borne encephalitis virus (TBEV) and ultrastructural changes induced by infection in the neuroblasts cell line (UKF-NB-4) was studied by electron microscopy. I compared electron microscopical aspects (appearance) of TEM images of neuroblasts cells prepared by (1) conventional chemical fixation, resin-embedding and sectionig; (2) rapid freezing of cell monolayers at high pressure and sectioning of freeze substituted samples. The most interesting fact, however, is that vitrification preserves the cell in close to native state, whereas chemical fixation and dehydration can not take place without extensive intra- and intermolecular cross-linking and aggregation. The appearance of the cytoplasm and nucleoplasm of neu-roblasts cells were different in conditions (1) and (2). The excellent ultrastructure of the cytoplasmic and nuclear membranes and organels of neuroblasts cells processed by (2) confirmed the potentional of the method for preservation of cellular fine structures. The infection of neuroblastoma cells was associated with number of major morphological changes, including proliferation of membranes of the rough endoplasmic reticulum and rearrangement of cytoskeletal structures. The viral particles were located mainly in the cisterna´s of ER but also in the cytoplasm. In the cytoplasm I observed virions in the asso-ciations with microtubules and neurosecretory dense core vesicles. The transport of viral particles inside of the transport vesicles was obsereved from ER to Golgi apparatus. Free nucleocapsids were not confirmed. The observed pattern corresponded to both trans and cis type of maturation. The TBEV-infected neuroblasts cells exhibited either apoptotic or necrotic morphological changes. I observed the apoptotic signs (condensation, margination and fragmentation of chromatin in nucleus) and other alterations, such as disorganisation of cytoplasm, presence of the vacuoles and high density of cytoplasm. This report also de-scribes scanning electron microscope study of the surface features of neuroblasts cells. We observed virus-mediated cytopathic effect. The cells infected with TBEV were rounded with rough and rugged topography.
| Identifer | oai:union.ndltd.org:nusl.cz/oai:invenio.nusl.cz:53235 |
| Date | January 2010 |
| Creators | TESAŘOVÁ, Martina |
| Source Sets | Czech ETDs |
| Language | Czech |
| Detected Language | English |
| Type | info:eu-repo/semantics/masterThesis |
| Rights | info:eu-repo/semantics/restrictedAccess |
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