A2-Astrocyte Activation by Short-Term Hypoxia Rescues α-Synuclein Pre-Formed-Fibril-Induced Neuronal Cell Death
Abstract
:1. Introduction
2. Materials and Methods
2.1. Animal
2.2. Primary Cortical Neuronal Culture
2.3. Primary Astrocyte Culture
2.4. Preparation of α-Syn PFF
2.5. Preparation of Hypoxia
2.6. qPCR Analysis
2.7. TUNEL Staining/Assay
2.8. Cell Viability Assay
2.9. AlamarBlue Assay
2.10. Immunofluorescence Analysis
2.11. Western Blot Analysis
2.12. Statistics
3. Results
3.1. Long-Term Hypoxia Induced Cell Death of Astrocytes
3.2. Short-Term Hypoxia Activated Astrocytes Without Cell Death
3.3. Short-Term Hypoxia Had a Protective Effect on Pre-Formed Fibril (PFF)-Treated Neurons
3.4. Hypoxia-Exposed Astrocyte-Conditioned Medium (hACM) Rescued PFF-Treated Neuronal Cell Death
3.5. Short-Term Hypoxia Provided a Neuro-Protective Effect by Promoting A2 Astrocytes and Suppressing A1 Astrocytes
4. Discussion
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Antibodies | Company | Catalog No. | Concentration |
---|---|---|---|
p-α-synuclein (Ser129) | Biolegend (San Diego, CA, USA) | 825701 | IF 1:1000 |
α-synuclein | BD Bioscience (Herlev, Denmark) | 610787 | WB 1:1000 |
β-actin-HRP | Sigma-Aldrich (St. Louis, MO, USA) | A3854 | WB 1:20,000 |
GFAP | EMD Millipore (Burlington, MA, USA) Dako (Santa Clara, CA, USA) | MAB360 Z033429 | WB 1:1000 IF 1:500 |
Tuj-1 | Biolegend (San Diego, CA, USA) | 802001 | IF 1:1000 |
HIF-1α | Cell Signaling (St. Louis, MO, USA) | 14179 | WB 1:500 |
CyTM3 AffiniPure Donkey Anti-Rabbit IgG (H + L) | Jackson (West Grove, PA, USA) | 711-165-152 | IF 1:500 |
Fluorescein (FITC) AffiniPure Donkey Anti-Mouse IgG (H + L) | Jackson | 715-095-151 | IF 1:500 |
Goat anti-Mouse IgG-heavy- and light-chain Antibody HRP Conjugated | BETHYL (Montgomery, TX, USA) | A90-116P | WB 1:10,000 |
Peroxidase-AffiniPure Goat Anti-Rabbit IgG (H + L) | Jackson | 111-035-144 | WB 1:10,000 |
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Choi, H.N.; Kim, S.-H.; Jo, M.G.; Lee, B.; Kim, Y.J.; Lee, S.E.; Lee, J.H.; Seong, H.M.; Kim, S.J.; Park, S.W.; et al. A2-Astrocyte Activation by Short-Term Hypoxia Rescues α-Synuclein Pre-Formed-Fibril-Induced Neuronal Cell Death. Biomedicines 2025, 13, 604. https://doi.org/10.3390/biomedicines13030604
Choi HN, Kim S-H, Jo MG, Lee B, Kim YJ, Lee SE, Lee JH, Seong HM, Kim SJ, Park SW, et al. A2-Astrocyte Activation by Short-Term Hypoxia Rescues α-Synuclein Pre-Formed-Fibril-Induced Neuronal Cell Death. Biomedicines. 2025; 13(3):604. https://doi.org/10.3390/biomedicines13030604
Chicago/Turabian StyleChoi, Ha Nyeoung, Seon-Hee Kim, Min Gi Jo, Bina Lee, Young Jin Kim, So Eun Lee, Jeong Hyun Lee, Hye Min Seong, Seong Jae Kim, Sang Won Park, and et al. 2025. "A2-Astrocyte Activation by Short-Term Hypoxia Rescues α-Synuclein Pre-Formed-Fibril-Induced Neuronal Cell Death" Biomedicines 13, no. 3: 604. https://doi.org/10.3390/biomedicines13030604
APA StyleChoi, H. N., Kim, S.-H., Jo, M. G., Lee, B., Kim, Y. J., Lee, S. E., Lee, J. H., Seong, H. M., Kim, S. J., Park, S. W., Kim, H. J., Kang, H., Lee, C. H., Lee, M. Y., Yun, S. P., & Kim, M. (2025). A2-Astrocyte Activation by Short-Term Hypoxia Rescues α-Synuclein Pre-Formed-Fibril-Induced Neuronal Cell Death. Biomedicines, 13(3), 604. https://doi.org/10.3390/biomedicines13030604