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Title: Microglial clearance function: dependence on phenotypes
Author: Oliveira, Ana Filipa Martins
Advisor: Brites, Dora
Jesus, Ana
Keywords: Unconjugated bilirubin
Microglia morphology
Cell migration
Nuclear factor-kappaB
Glycoursodeoxycholic acid
Defense Date: 2011
Publisher: Faculdade de Ciências e Tecnologia
Abstract: Microglia are active sensors of the brain and respond promptly to even minor disturbance in their microenvironment. A feature of this response is the accumulation of these cells at the site of lesion. Neonatal jaundice is a common condition of the newborn and may determine injury to neurons and glial cells, such as microglia, when levels of unconjugated bilirubin (UCB) are excessive. With the objective to evaluate whether microglia have a protective or deleterious role, we decided to assess, using the Boyden chamber, the chemotactic effect of free unbound UCB (fUCB), as well as the migration ability of UCB-treated microglia in the absence or in the presence of chemotatic compounds, such as ATP and S100B. Also, we intended to evaluate the effect of glycoursodeoxycholic acid (GUDCA) as a modulator. To characterize our usual model of microglia isolation, phenotypic evaluation of cultures with different days in vitro (DIV) was performed by estimating cell morphology, nuclear factor-kappaB (NF-κB) activation and phagocytic ability. We observed that fUCB did not act as a chemotactic compound for microglia and that cells treated with UCB showed decreased migration ability. Co-incubation with GUDCA prevented this effect and enhanced microglia migration. However, reduced effects were observed in the presence of ATP and abolished when using S100B. Isolated microglia with 2 DIV showed features of activation, but presentedramified morphology of the “resting” state, less NF-κB activation and increased phagocytosis at 13 DIV. Data indicate that microoglia exposure to UCB leads to a reduced migration ability and that co-incubation with GUDCA prevents this deleterious effect, resulting in an increased migration. Characterization of microglia phenotypes, along the time in culture, point to 13 DIV cells as the most suitable for studies intended to evaluate microglia reactivity to UCB, and probably to other stimuli.
Description: Dissertação para obtenção do Grau de Mestre em Genética Molecular e Biomedicina
Appears in Collections:FCT: DCV - Dissertações de Mestrado

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