Title Page
Contents
국문초록 8
ABSTRACT 10
I. Introduction 12
II. Objectives 15
III. Material and Methods 16
A. Isolation of primary cortical cells 16
B. Three-dimensional neural spheroid culture 16
C. Immunofluorescence 17
D. Cell viability 18
E. Measurement of BDNF mRNA levels by qRT-PCR 19
F. Neurite outgrowth analysis 20
G. Western blotting 20
H. Statistical analysis 21
IV. Results 24
A. Self-assembled 3D spheroids in cortical cells 24
B. Changes in cell viability, BDNF mRNA level, and neurite outgrowth in neural spheroids after DEX exposure 24
C. Changes in the levels of BDNF and sirtuin 1 proteins and ERK1/2 phosphorylation in neural spheroids after DEX exposure 25
C. Changes in the phosphorylation of mTORC1-mediated proteins and the expression of synaptic proteins in neural spheroids after DEX exposure 26
V. Discussion 40
VI. Conclusion 48
References 49
Table 1. List of antibodies used in this study. 22
Figure 1. 3D cortical spheroid fabrication. 28
Figure 2. Cortical spheroids with neural cell types. 30
Figure 3. Changes in cell viability, BDNF mRNA level, and neurite outgrowth after dexamethasone (DEX) exposure. 33
Figure 4. Changes in the levels of BDNF and sirtuin 1 proteins and ERK1/2 phosphorylation after dexamethasone (DEX) exposure. 36
Figure 5. Changes in the phosphorylation of mTORC1-mediated proteins and the expression of synaptic proteins after dexamethasone (DEX) exposure. 39
Figure 6. Schematic diagram of the molecular mechanisms underlying DEX-induced impaired neuroplasticity. 46