Publicação
Betacellulin and Neurogenesis in the Adult Central Nervous System
| Resumo: | Neural stem cells (NSCs) reside in special niches in the adult brain, including subventricular zone (SVZ) of the lateral ventricle and subgranular zone (SGZ) of the dentate gyrus. Blood vessels are an important coumpound of the neurogenic niches as they secreate proteins such as betacellulin (BTC) that stimulate NSC proliferation, self-‐ renewal and differentiation. BTC is a member of the epidermal growth factor (EGF) family of ligands, and has been widely studied in many different contexts. Recently, it has been demonstrated that, in vitro, BTC can induce NSC proliferation, promote self-‐renewal and prevent spontaneous differentiation. In vivo, BTC can also promote neurogenesis. BTC is released into the neurogenic niche by endothelial cells of the microvasculature and by the choroid plexus (CP). It is thought that BTC activity in NSCs is mediated through ErbB1 and ErbB4 receptors whose activation stimulate the AKT and MEK signalling pathways. In my thesis I carried out analysis understanding the influence of BTC and other growth factors on NSCs in vitro using the neurosphere assay and measuring neurospheres number and size. I also studied the importance of AKT and MEK signalling pathways in NSCs cultivated in medium supplemented with different growth factors including BTC. I observed an increase in cell cycle arrest when inhibitors of both signalling pathways were added together in all culture conditions. Studies to better characterize the interaction between neurogenic niche and BTC were also carried out using newly developed visualisation techniques, SeeDB and CLARITY that allowed us to have a 3D perspective. In parallel, I made BTC conditional and BTC reporter contructs for generating mice using newly developed CRISPR/ Cas 9 technique. These mice will allow us to better understand parallel, I made BTC conditional and BTC reporter contructs for generating mice using newly developed CRISPR/ Cas 9 technique. These mice will allow us to better understand the relative importance of BTC in adult NSCs and whether BTC transcription is modulated |
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| Autores principais: | Santos, Inês da Costa |
| Assunto: | Neurogénese adulta ZSV Betacelulina SeeDB Clarity CRISPR/CAS 9 |
| Ano: | 2015 |
| País: | Portugal |
| Tipo de documento: | dissertação de mestrado |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade de Coimbra |
| Idioma: | inglês |
| Origem: | Estudo Geral - Universidade de Coimbra |
| Resumo: | Neural stem cells (NSCs) reside in special niches in the adult brain, including subventricular zone (SVZ) of the lateral ventricle and subgranular zone (SGZ) of the dentate gyrus. Blood vessels are an important coumpound of the neurogenic niches as they secreate proteins such as betacellulin (BTC) that stimulate NSC proliferation, self-‐ renewal and differentiation. BTC is a member of the epidermal growth factor (EGF) family of ligands, and has been widely studied in many different contexts. Recently, it has been demonstrated that, in vitro, BTC can induce NSC proliferation, promote self-‐renewal and prevent spontaneous differentiation. In vivo, BTC can also promote neurogenesis. BTC is released into the neurogenic niche by endothelial cells of the microvasculature and by the choroid plexus (CP). It is thought that BTC activity in NSCs is mediated through ErbB1 and ErbB4 receptors whose activation stimulate the AKT and MEK signalling pathways. In my thesis I carried out analysis understanding the influence of BTC and other growth factors on NSCs in vitro using the neurosphere assay and measuring neurospheres number and size. I also studied the importance of AKT and MEK signalling pathways in NSCs cultivated in medium supplemented with different growth factors including BTC. I observed an increase in cell cycle arrest when inhibitors of both signalling pathways were added together in all culture conditions. Studies to better characterize the interaction between neurogenic niche and BTC were also carried out using newly developed visualisation techniques, SeeDB and CLARITY that allowed us to have a 3D perspective. In parallel, I made BTC conditional and BTC reporter contructs for generating mice using newly developed CRISPR/ Cas 9 technique. These mice will allow us to better understand parallel, I made BTC conditional and BTC reporter contructs for generating mice using newly developed CRISPR/ Cas 9 technique. These mice will allow us to better understand the relative importance of BTC in adult NSCs and whether BTC transcription is modulated |
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