Estudi de la unió de la toxina èpsilon de "Clostridium perfringens" a diferents teixits i el seu pas a través de la barrera hematoencefàlica

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(1)Estudi de la unió de la toxina èpsilon de Clostridium perfringens a diferents teixits i el seu pas a través de la barrera hematoencefálica Jonatan Dorca Arévalo. ADVERTIMENT. La consulta d’aquesta tesi queda condicionada a l’acceptació de les següents condicions d'ús: La difusió d’aquesta tesi per mitjà del servei TDX (www.tdx.cat) ha estat autoritzada pels titulars dels drets de propietat intel·lectual únicament per a usos privats emmarcats en activitats d’investigació i docència. No s’autoritza la seva reproducció amb finalitats de lucre ni la seva difusió i posada a disposició des d’un lloc aliè al servei TDX. No s’autoritza la presentació del seu contingut en una finestra o marc aliè a TDX (framing). Aquesta reserva de drets afecta tant al resum de presentació de la tesi com als seus continguts. En la utilització o cita de parts de la tesi és obligat indicar el nom de la persona autora.. ADVERTENCIA. La consulta de esta tesis queda condicionada a la aceptación de las siguientes condiciones de uso: La difusión de esta tesis por medio del servicio TDR (www.tdx.cat) ha sido autorizada por los titulares de los derechos de propiedad intelectual únicamente para usos privados enmarcados en actividades de investigación y docencia. No se autoriza su reproducción con finalidades de lucro ni su difusión y puesta a disposición desde un sitio ajeno al servicio TDR. No se autoriza la presentación de su contenido en una ventana o marco ajeno a TDR (framing). Esta reserva de derechos afecta tanto al resumen de presentación de la tesis como a sus contenidos. En la utilización o cita de partes de la tesis es obligado indicar el nombre de la persona autora.. WARNING. On having consulted this thesis you’re accepting the following use conditions: Spreading this thesis by the TDX (www.tdx.cat) service has been authorized by the titular of the intellectual property rights only for private uses placed in investigation and teaching activities. Reproduction with lucrative aims is not authorized neither its spreading and availability from a site foreign to the TDX service. Introducing its content in a window or frame foreign to the TDX service is not authorized (framing). This rights affect to the presentation summary of the thesis as well as to its contents. In the using or citation of parts of the thesis it’s obliged to indicate the name of the author..

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Fig. 1. Schematic drawing of native and recombinant ewere generated as fusion proteins with the GST (from pGEX-4T-1 vector), with or without GFP, and lacking the N-terminal peptide(remove the GST (black arrows) and the resultingproteolytic cleavage of N- a
Fig. 1. Schematic drawing of native and recombinant ewere generated as fusion proteins with the GST (from pGEX-4T-1 vector), with or without GFP, and lacking the N-terminal peptide(remove the GST (black arrows) and the resultingproteolytic cleavage of N- a p.226
Fig. 2. Localisation of e-prototoxin–GFP and e-toxin–GFP in brain after i.v. injection in mice
Fig. 2. Localisation of e-prototoxin–GFP and e-toxin–GFP in brain after i.v. injection in mice p.228
Fig. 3. Localisation of Alexa 546-labelled BSA in blood vessels after einjected mice BSA remained confined to the blood vessel’s lumen (C, E), in the case ofi.v
Fig. 3. Localisation of Alexa 546-labelled BSA in blood vessels after einjected mice BSA remained confined to the blood vessel’s lumen (C, E), in the case ofi.v p.229
Fig. 4. Localisation of e-toxin–GFP on glial brain cells after i.v. injection in mice
Fig. 4. Localisation of e-toxin–GFP on glial brain cells after i.v. injection in mice p.230
Fig. 5. Cytotoxic effect of e-toxin on glial cells. MTT cell deathassays were performed on mixed glial primary cell cultures byexposing the cells to 250 nM e-prototoxin (open circles), e-toxin at5 nM (black triangles), 50 nM (black squares) or 250 nM (blac
Fig. 5. Cytotoxic effect of e-toxin on glial cells. MTT cell deathassays were performed on mixed glial primary cell cultures byexposing the cells to 250 nM e-prototoxin (open circles), e-toxin at5 nM (black triangles), 50 nM (black squares) or 250 nM (blac p.231
Fig. 6. Identification of the cell type killed by e(G, I, K) and immunofluorescence against GFAP for astrocytes (H, J, L) were performed
Fig. 6. Identification of the cell type killed by e(G, I, K) and immunofluorescence against GFAP for astrocytes (H, J, L) were performed p.232
Fig. 1. Glutamate release from mouse brain synaptosomes. Gluta-mate release from mouse brain synaptosomes was measured on lineby a spectrofluorimetric method (see Section 2)
Fig. 1. Glutamate release from mouse brain synaptosomes. Gluta-mate release from mouse brain synaptosomes was measured on lineby a spectrofluorimetric method (see Section 2) p.241
Fig. 2B and E, respectively) were used to stain isolated
Fig. 2B and E, respectively) were used to stain isolated p.242
Fig. 3. Binding of e-toxin to myelinic structures in mouse brain. Sections from mouse cerebellum were incubated with e-toxin-GFP (A and C)
Fig. 3. Binding of e-toxin to myelinic structures in mouse brain. Sections from mouse cerebellum were incubated with e-toxin-GFP (A and C) p.243
Fig. 4. Effect of pronase E, N-glycosidase F and Triton X-100 on the e-prototoxin-GFP binding to myelin
Fig. 4. Effect of pronase E, N-glycosidase F and Triton X-100 on the e-prototoxin-GFP binding to myelin p.244
Fig. 5. Binding of e-toxin to peripheral nerve fibres. (A) Cross-section of mouse left vagus nerve incubated with e-prototoxin-GFP.(B) Teased fibre from mouse sciatic nerve incubated with e-proto-
Fig. 5. Binding of e-toxin to peripheral nerve fibres. (A) Cross-section of mouse left vagus nerve incubated with e-prototoxin-GFP.(B) Teased fibre from mouse sciatic nerve incubated with e-proto- p.245
Fig. 6. Binding of eDetail of a section from sheep striatum showing the binding oflayer; PL: Purkinje cell layer and ML: molecular layer
Fig. 6. Binding of eDetail of a section from sheep striatum showing the binding oflayer; PL: Purkinje cell layer and ML: molecular layer p.246
Table 1Click here to download high resolution image

Table 1Click

here to download high resolution image p.277
Table 2Click here to download high resolution image

Table 2Click

here to download high resolution image p.278
Figure 1Click here to download high resolution image

Figure 1Click

here to download high resolution image p.279
Figure 2Click here to download high resolution image

Figure 2Click

here to download high resolution image p.280
Figure 3Click here to download high resolution image

Figure 3Click

here to download high resolution image p.281
Figure 4Click here to download high resolution image

Figure 4Click

here to download high resolution image p.282
Figure 5Click here to download high resolution image

Figure 5Click

here to download high resolution image p.283
Figure 6Click here to download high resolution image

Figure 6Click

here to download high resolution image p.284

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