Characterization of Notch1, Notch2 and Rbpj roles in the generation of neural stem cells derived from reactive astrocytes

Abstract

Astrocytes are activated by injuries to the central nervous system (CNS) and acquire an undifferentiated state, which is characteristic of neural stem cells (NSCs). In vitro these astrocyte-derived NSC-like cells regain multipotency and long-term self-renew, however, the molecular factors involved in astrocyte dedifferentiation are poorly understood. The purpose of this research project is to study the role of the Notch signaling pathway in the generation of NSCs from reactive astrocytes in vitro. We will induce conditional Notch1, Notch2 and Rbpj ablations from astrocytes in the adult brain to study the separate roles of Notch pathway components on the ability of reactive astrocyte to dedifferentiate. The mutant astrocytes will be isolated from the cortex of adult mice by fluorescence assisted cell sorting (FACS) and cultured in vitro. Reactivity will be inducted by mechanical lesion and reactive astrocytes-derived NSC will be assessed in their proliferative and differentiation potential by immunostaining and RT-qPCR assays. (AU)