Indução de diferenciação causa mudança na localização subcelular de Runx em células-tronco neurais

• Ano de defesa: 2015
• Autor(a) principal: Massinhani, Fernando Henrique [UNIFESP]
• Orientador(a): Porcionatto, Marimelia Aparecida [UNIFESP]
• Tipo de documento: Dissertação
• Tipo de acesso: Acesso aberto
• dARK ID: ark:/48912/001300001rqjb
• Idioma: por
• Instituição de defesa: Universidade Federal de São Paulo (UNIFESP)
• Palavras-chave em Português: Células-tronco neurais; Diferenciação celular; Fatores de transcrição; Runx
• Link de acesso: http://repositorio.unifesp.br/handle/11600/48252; https://sucupira.capes.gov.br/sucupira/public/consultas/coleta/trabalhoConclusao/viewTrabalhoConclusao.jsf?popup=true&id_trabalho=2357930

Runx family members are transcription factors evolutionarily conserved that regulate the expression of genes involved in cell differentiation and cell cycle progression, that may act as transcriptional activators or repressors. The aim of this thesis was to verify whether the expression of the Runx family members (Runx1, 2 and 3) by neural stem cells (NSC) derived from adult mice subventricular zone (SVZ) is related to the differentiation of these cells into neurons. Using NSC cultured as neurospheres we observed Runx family members gene and protein expression by PCR and Western blotting respectively. In order to analyze the subcellular localization of RUNX, neurospheres were induced to adhere to the substrate and were maintained in culture for 1, 7 and 14 days to promote differentiation. Finally, we investigated the presence of Delta-Notch pathway components in the SVZ and their location relative to RUNX3 neural stem cell. By immunocytochemistry we observed that RUNX-1, -2 and -3 exhibited similar pattern of localization, being found predominantly in the nucleus of the cells cultured up to 7 days and in the cytoplasm after 14 days. RUNX3 was localized in different cell types, identified by specific markers for NSC, neuroblasts and neurons. Our results show predominance of nuclear localization after 7 days in vitro (d.i.v) and cytoplasmic expression after 14 d.i.v, regardless of cell type. We also identified the presence of HES1, TLE1 and NOTCH1 in the SVZ which colocalized with RUNX3 in neural stem cells. So far, our results suggest that Runx family members are expressed by NSC in vitro and that the localization and expression are probably related to the stage of differentiation of these cells into neurons. Furthermore, we observed that the members of Notch-Delta (HES1, TLE1 and Notch cytoplasmic portion) have the same subcellular location which RUNX3.