Tese

Ativação microglial, lesão da substância branca e expressão de Nogo-A em ratos submetidos à isquemia estriatal

The objective of this investigation was to evaluate the degenerative pattern of several white matter tracts after striatal ischemic injury, correlating degenerative process standards with the microglial activation and expression of Nogo-A. For this purpose, focal ischemia was induced with stereotact...

ver descrição completa

Autor principal: LIMA, Rafael Rodrigues
Grau: Tese
Idioma: por
Publicado em: Universidade Federal do Pará 2012
Assuntos:
Acesso em linha: http://repositorio.ufpa.br/jspui/handle/2011/2996
Resumo:
The objective of this investigation was to evaluate the degenerative pattern of several white matter tracts after striatal ischemic injury, correlating degenerative process standards with the microglial activation and expression of Nogo-A. For this purpose, focal ischemia was induced with stereotactic injection of endothelin in striatum of adult rats, and only in the control animals injected with sterile saline. The animals were perfused 3, 7, 14 and 30 days after ischemia. The brain removed, postfixed, cryoprotected, cut into cryostat sections obtained and submitted to immunohistochemical investigation with the following antibodies: anti-GFAP (1:2000, Dako), anti-Tau-1 (1:500, Chemicon), Anti-MBP (1:100, Chemicon International), Anti-Nogo-A (1:100, Invitrogen), Anti-Iba1 (1:1000, WAKO), ED1 (1:500, Serotec) and Anti-MHC II (Abcam 1:100), besides the viewing of the damage pattern with cresyl violet. Slides are marked by different methods were evaluated qualitatively and quantitatively also some (Anti-Nogo A, anti-ED-1, anti-MHC-II and anti-tau-1), counts were carried out in the striatum and in the corpus callosum. The data were tabulated, statistically analyzed by Tukey test (p <0.05) and micrographs taken of the findings more representative. The slides were stained with cresyl violet revealed an increase in cell density by the infiltration of inflammatory cells to the ischemic area, with a significant increase on day 7. The blades immunostained for GFAP was found progressive increase of the population of astrocytes and an increase in cell volume 7 and 14 days. Oligodendrocyte pathology marked with Tau-1 had peak marking the 3rd day in the striatum and the 7th day in the corpus callosum, and loss of myelin compaction identified by MBP was better at 14, in the different treatment. The microglial activation identified by different immunoblots showed a peak on day 7, both in striatum and in the corpus callosum, but in the corpus callosum with a much smaller number compared to the striatum. The morphology of microglial underwent changes, which found the branched phenotype in control animals, as well as in early and late times after ischemia and amoeboid default / phagocytic day 7, coinciding with the largest number of activated cells. The count of Nogo-A + cells peaked at 3 days observed in the striatum, and there were no differences in the corpus callosum expression Nogo-A 3 to 14 days, only a decrease compared to 30 days. Thus, microinjections of ET-1 induced conspicuous striatal tissue loss, concomitant with progressive microglial activation, astrocytosis, loss of immunoreactivity for myelin basic protein and oligodendrocytes damage in various survival times after focal ischemia. These events affect a few SB tracts, as the corpus callosum. The establishment of the temporal evolution of these events is the neuropathological basis for future studies, in which they should handle the inflammatory response in order to minimize these tissue changes.