Neural stem cells (NSCs) in the brain have the ability to differentiate into neurons, oligodendrocytes and strocytes. Autophagy is a major degradation pathway for cellular homeostasis and remodeling. However, little is known how autophagy is associated with differentiation of NSCs. This study was aimed to examine whether autophagy plays a critical role for self-renewal and differentiation of adult hippocampal neural stem (HCN) cells. Suppression of autophagy by knockdown of Atg7, a key autophagy gene, impaired differentiation of HCN cells into neuron, oligodendrocyte and astrocyte. After induction of differentiation of HCN cells to astrocytes by retinoic acid, considerable amount of autophagosomes was observed in the early time point, but later autophagy level returned to the basal state. Recent studies suggested that astrocytes are dynamic regulators of neuronal activity such as in neuronal development, activity and differentiation. Our findings suggest that autophagy plays a key role in regulation of NSC differentiation, and especially astrogenesis depends on autophagy. Our study will help to elucidate the importance of autophagy in regulation of HCN cell differentiation. ⓒ 2017 DGIST
Table Of Contents
1. Introduction 1-- 2. Materials and methods 8-- 2.1 Primary culture of adult rat HCN cells 8-- 2.2 Media for maintenance and differentiation of HCN cells 8-- 2.3 Generation of stable cell line 9-- 2.4 RNA extraction and qRT-PCR 10-- 2.5 Immunocytochemistry 12-- 2.6 Western blotting 12-- 3. Results 14-- 3.1 HCN cells show intact differentiation ability into neurons, oligodendrocytes and astrocytes 14-- 3.2 Characterization of HCN cells differentiation into astrocytes 17-- 3.3 Autophagy is highly activated in the early stage of differentiation and decreased later 20 -- 3.4 Atg7 stable knockdown impairs astrogenesis 22-- 4. Discussion 26-- Reference 29