Store-Operated Calcium Entry Inhibition and Plasma Membrane Calcium Pump Upregulation Contribute to the Maintenance of Resting Cytosolic Calcium Concentration in A1-like Astrocytes
Abstract
:1. Introduction
2. Results
2.1. Selection of A1-like Astrocytes Using C3 Expression as Biomarker
2.2. A1-like Astrocytes Express Higher Levels of Aβ Peptides Than Control U251 Astrocytes
2.3. Store-Operated Calcium Entry (SOCE) Is Largely Decreased during the Transformation of U251 Astrocytes in A1-like Astrocytes
2.4. The Cytosolic Ca2+ Concentration Does Not Increase during the Transformation of U251 Astrocytes in A1-like Astrocytes, Which Express Higher Levels of PMCA Than the Control U251 Astrocytes
3. Discussion
4. Materials and Methods
4.1. Materials
4.2. Cell Culture and Preparation of Reactive Astrocytes
4.3. U251 Cell Staining with C3 and Aβ Antibodies
4.4. Cell Homogenate Preparation, and Western Blot and Dot Blot Analyses
4.5. Intracellular Cytosolic Ca2+ Measurements
4.6. Statistics
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
Sample Availability
Abbreviations
Aβ | amyloid β peptide |
AD | Alzheimer’s disease |
APP | amyloid β protein precursor |
BACE1 | β-site APP-cleaving enzyme 1 |
C1q | complement component 1q |
DMEM | Dulbecco’s modified Eagle’s medium |
EGTA | ethyleneglycoltetraacetic acid |
ER | endoplasmic reticulum |
Fluo3 AM | Fluo-3-pentaacetoxymethyl ester |
Fura2 AM | Fura2 acetoxymethyl ester |
GAPDH | glyceraldehyde-3-phosphate dehydrogenase |
GF | green fluorescence |
HEPES | N-[2-hydroxyethyl] piperazine-N′-[2-ethanesulfonic acid] |
IL-1α | interleukin-1α |
NPA | 3-nitropropionic acid |
PBS | phosphate-buffered saline |
PBST | PBS supplemented with 0.2% 4-(1,1,3,3-tetramethyl butyl) phenyl-polyethylene glycol (Triton X-100®) |
PMCA | plasma membrane calcium pump |
PSEN | presenilin |
PVDF | polyvinylidene difluoride |
RF | red fluorescence |
ROI | region of interest |
SDS-PAGE | sodium dodecyl sulfate-polyacrylamide gel electrophoresis |
SEM | standard error of the mean |
SOCE | store-operated calcium entry |
STIM1 | stromal interaction molecule 1 |
Tg | thapsigargin |
TNFα | tumor necrosis factor α |
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Poejo, J.; Berrocal, M.; Saez, L.; Gutierrez-Merino, C.; Mata, A.M. Store-Operated Calcium Entry Inhibition and Plasma Membrane Calcium Pump Upregulation Contribute to the Maintenance of Resting Cytosolic Calcium Concentration in A1-like Astrocytes. Molecules 2023, 28, 5363. https://doi.org/10.3390/molecules28145363
Poejo J, Berrocal M, Saez L, Gutierrez-Merino C, Mata AM. Store-Operated Calcium Entry Inhibition and Plasma Membrane Calcium Pump Upregulation Contribute to the Maintenance of Resting Cytosolic Calcium Concentration in A1-like Astrocytes. Molecules. 2023; 28(14):5363. https://doi.org/10.3390/molecules28145363
Chicago/Turabian StylePoejo, Joana, María Berrocal, Lucía Saez, Carlos Gutierrez-Merino, and Ana M. Mata. 2023. "Store-Operated Calcium Entry Inhibition and Plasma Membrane Calcium Pump Upregulation Contribute to the Maintenance of Resting Cytosolic Calcium Concentration in A1-like Astrocytes" Molecules 28, no. 14: 5363. https://doi.org/10.3390/molecules28145363