Abstract
Excessive activation of α-amino-3-hydroxy-5-methyl-4-isoxazole propoinic acid (AMPA) receptors instigates excitotoxicity via enhanced calcium influx in the neurons thus inciting deleterious consequences. Additionally, Endoplasmic Reticulum (ER) is pivotal in maintaining the intracellular calcium balance. Considering this, studying the aftermath of enhanced calcium uptake by neurons and its effect on ER environment can assist in delineating the pathophysiological events incurred by excitotoxicty. The current study was premeditated to decipher the role of ER pertaining to calcium homeostasis in AMPA-induced excitotoxicity. The findings showed, increased intracellular calcium levels (measured by flowcytometry and spectroflourimeter using Fura 2AM) in AMPA excitotoxic animals (male Sprague dawely rats) (intra-hippocampal injection of 10 mM AMPA). Further, ER resident proteins like calnexin, PDI and ERp72 were found to be upregulated, which further modulated the functioning of ER membrane calcium channels viz. IP3R, RyR, and SERCA pump. Altered calcium homeostasis further led to ER stress and deranged the protein folding capacity of ER post AMPA toxicity, which was ascertained by unfolded protein response (UPR) pathway markers such as IRE1α, eIF2α, and ATF6α. Chemical chaperone, 4-phenybutric acid (4-PBA), ameliorated the protein folding capacity and subsequent UPR markers. In addition, modulation of calcium channels and calcium regulating machinery of ER post 4-PBA administration restored the calcium homeostasis. Therefore the study reinforces the significance of ER stress, a debilitating outcome of impaired calcium homeostasis, under AMPA-induced excitotoxicity. Also, employing chaperone-based therapeutic approach to curb ER stress can restore the calcium imbalance in the neuropathological diseases.
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Data Availability
All data generated or analyzed during this study are included in this manuscript. Primary datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- AMPA:
-
α-Amino-3-hydroxy-5-methyl-4-isoxazole propoinic acid
- ATF-6:
-
Activating transcription factor-6
- ER:
-
Endoplasmic reticulum
- GRP75:
-
Glucose regulated protein75
- IP3R:
-
Inositol 1,4,5-triphosphate receptors
- KA:
-
Kianic acid
- MAM:
-
Mitochondrial-associated membrane
- NMDA:
-
N-methyl-D-aspartate
- 4-PBA:
-
4-Phenylbutyric acid
- RyR:
-
Ryanodine receptors
- SERCA:
-
Sacroendoplasmic reticulum Ca ATPase
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Acknowledgements
We would like to thank center facilities instrumentation laboratory of Centre for Stem Cell and Tissue Engineering and Excellence in Biomedical Sciences Panjab University (PU) Chandigarh for providing facilities for stereotactic surgeries.
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This work was supported by the University Grants Commission-Basic Scientific Research (UGC-BSR) (F.25–1/2013–14(BSR)/7–209/2009(BSR), Department of Science and Technology (FIST) (SR/FST/LS1-645) and University Grant Commission (SAP) (F.4–1/2015/DSA-1 (Sap-4).
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Study concept and design were elaborated by AB and TK. Material preparations and data collection were performed by AB. Formal analysis was accomplished by AB, RB, and TK. The first draft of the manuscript was written by AB and TK supervised the study. All the authors AB, RB, AS, DKD and TK reviewed and commented on the previous version of manuscript. All the authors have read and approved the final manuscript.
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Bhardwaj, A., Bhardwaj, R., Saini, A. et al. Impact of Calcium Influx on Endoplasmic Reticulum in Excitotoxic Neurons: Role of Chemical Chaperone 4-PBA. Cell Mol Neurobiol 43, 1619–1635 (2023). https://doi.org/10.1007/s10571-022-01271-y
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DOI: https://doi.org/10.1007/s10571-022-01271-y