Development of p-Tau Differentiated Cell Model of Alzheimer’s Disease to Screen Novel Acetylcholinesterase Inhibitors
Abstract
:1. Introduction
2. Results
2.1. GA Treatment Induces Abnormal Tau Phosphorylation and Axon Degeneration in SH-SY5Y-Differentiated Neurons
2.2. Novel Compounds Inhibit AChE Enzyme Activity in SH-SY5Y-Differentiated Neurons
2.3. AChE Inhibition Prevents GA-Induced Tau Phosphorylation on S396 but Not on S199
2.4. Dual AChE/GSK-3 Inhibitor 27g Modulates Tau Phosphorylation on Both S199 and S396
2.5. Dual AChE/GSK-3 Inhibitor 27g Prevents Tau-Induced Neurodegeneration and Protects from Neuronal Cell Death
3. Discussion
4. Materials and Methods
4.1. Reagents
4.2. SH-SY5Y Culture and Neuronal Differentiation
4.3. Glyceraldehyde Induced Tau Hyperphosphorylation
4.4. Immunostaining Cell Culture
4.5. Confocal Microscopy
4.6. pTAU Quantification
4.7. AChE Activity Assay (Ellman’s Method)
4.8. Cell Viability Assay
4.9. Data Analysis
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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No. | Compd. | Structures |
---|---|---|
A | XJP-1 | |
Name: (E)-4-((5,6-dimethoxy-1-oxo-1,3-dihydro-2H-inden-2-ylidene)methyl)-1-(3-fluorobenzyl)pyridin-1-ium bromide | ||
B | SAD-2 | |
Name: (Z)-4-((6,7-dimethoxy-4-oxoisothiochroman-3-ylidene)methyl)-1-(4-fluorobenzyl)pyridin-1-ium bromide | ||
C | SAD-6 | |
Name: (Z)-3-((1-benzylpiperidin-4-yl)methylene)-6,7-dimethoxyisothiochroman-4-one | ||
D | 24r | |
Name: 2-((1-(4-fluorobenzyl)piperidin-4-yl)methyl)-5,6-dimethoxy-2,3-dihydrobenzo[b]thiophene 1,1-dioxide | ||
E | 27g | |
Name: 2-((7-((6-chloro-1,2,3,4-tetrahydroacridin-9-yl)amino)heptyl)amino)-6-(3-fluoropyridin-4-yl)-3-methylpyrimidin-4(3H)-one |
Basic Growth Media | Differentiation Media #1 | Differentiation Media #2 | Differentiation Media #3 |
---|---|---|---|
EMEM | EMEM | EMEM | Neurobasal |
15% hiFBS | 2.5% hiFBS | 1% hiFBS | 20 mM KCl |
1x Pen/Strep | 1x Pen/Strep | 1x Pen/Strep | 1x Pen/Strep |
2 mM Glutamine | 2 mM Glutamine | 2 mM Glutamine | 2 mM Glutamax |
10 μM RA | 10 μM RA | 10 μM RA | |
50 ng/mL BDNF |
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Uras, G.; Li, X.; Manca, A.; Pantaleo, A.; Bo, M.; Xu, J.; Allen, S.; Zhu, Z. Development of p-Tau Differentiated Cell Model of Alzheimer’s Disease to Screen Novel Acetylcholinesterase Inhibitors. Int. J. Mol. Sci. 2022, 23, 14794. https://doi.org/10.3390/ijms232314794
Uras G, Li X, Manca A, Pantaleo A, Bo M, Xu J, Allen S, Zhu Z. Development of p-Tau Differentiated Cell Model of Alzheimer’s Disease to Screen Novel Acetylcholinesterase Inhibitors. International Journal of Molecular Sciences. 2022; 23(23):14794. https://doi.org/10.3390/ijms232314794
Chicago/Turabian StyleUras, Giuseppe, Xinuo Li, Alessia Manca, Antonella Pantaleo, Marco Bo, Jinyi Xu, Stephanie Allen, and Zheying Zhu. 2022. "Development of p-Tau Differentiated Cell Model of Alzheimer’s Disease to Screen Novel Acetylcholinesterase Inhibitors" International Journal of Molecular Sciences 23, no. 23: 14794. https://doi.org/10.3390/ijms232314794