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Title Page

ABSTRACT

Contents

I. INTRODUCTION 12

II. MATERIALS AND METHODS 20

A. Chemicals and antibodies 20

B. Constructs 20

C. Animals 22

D. Cell culture 23

E. Transfection 23

F. Tissue & Cell sample preparation 24

G. Immunoblot analysis 25

H. Immunohistochemistry 25

I. Immunocytochemistry 26

J. Reverse transcription polymerase chain reaction (RT-PCR) 27

K. Statistical analysis 28

III. RESULTS 29

1. Circadian rhythm was disrupted in P301S mutant human tau transgenic mice 29

2. Circadian disruption was observed also in the cortex and hippocampus of P301S tau transgenic mice 34

3. The status of pathologic tau was altered according to ZT 40

4. Oscillations of tau solubility was relevant to the chaperon Hsp70 protein 43

5. Bmal1 modulated tau status and stability via Hsp70 upregulation 48

6. Hsp70 was involved in the modulation of tau status and stability 54

IV. DISCUSSION 62

V. REFERENCES 70

국문 요약 76

List of Figures

Figure 1. The circadian rhythm-related physiology. 17

Figure 2. The molecular mechanism of circadian clock. 18

Figure 3. The formation of neurofibrillary tangles (NFTs) in tauopathies. 19

Figure 4. Full picture for drawing ROI of SCN. 31

Figure 5. Circadian rhythm expressions of master clock molecules in... 32

Figure 6. Transcription levels of circadian oscillators in diencephalon... 33

Figure 7. Disrupted rhythmicity of core clock genes in the cortex of... 37

Figure 8. Disrupted rhythmicity of core clock genes in the... 39

Figure 9. ZT-related alteration of pathologic tau status. 41

Figure 10. Densitometry analysis of pathologic tau status. 42

Figure 11. The diurnal alteration of Hsp70. 44

Figure 12. Oscillations of tau solubility relevant to the chaperon Hsp70 protein. 45

Figure 13. Reverse correlation between the insoluble tau and Hsp70. 46

Figure 14. Co-localization of Hsp70 and tau. 47

Figure 15. Tau-aggregation enhanced by forskolin treatment. 50

Figure 16. Increase in Hsp70 by overexpression of Bmal1. 51

Figure 17. Bmal1 regulates mutant tau status as well as its... 52

Figure 18. Degradation of mutant tau by Bmal1 was mediated by HSF1... 53

Figure 19. Hsp70-mediated tau disaggregation and following its degradation. 56

Figure 20. No alteration of Tau mRNA levels by Hsp70. 57

Figure 21. Co-localization of Hsp70 and tau in tau overexpressing... 58

Figure 22. Involvement of autophagy and proteasome in Hsp70-... 59

Figure 23. The regulation of tau degradation by Bmal1-Hsp70. 60

Figure 24. Disaggregation of tau prior to its degradation mediated by Hsp70. 61

Figure 25. Hypothetical model for the interrelation mechanism... 63

Figure 26. Prediction of Bmal1 binding on the promoter region of Hsp70. 65

Figure 27. Hsc70, a possible another downstream target of Bmal1. 66

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