P2X7 receptor activation mediates superoxide dismutase 1 (SOD1) release from murine NSC-34 motor neurons.

Abstract

Mutant superoxide dismutase 1 (SOD1) can be constitutively released from motor neurons and transmitted to naïve motor neurons to promote the progression of amyotrophic lateral sclerosis (ALS). However, the biological impacts of this process and the precise mechanisms of SOD1 release remain to be fully resolved. Using biochemical and fluorescent techniques, this study aimed to determine if P2X7 receptor activation could induce mutant SOD1 release from motor neurons and whether this released SOD1 could be transmitted to motor neurons or microglia to mediate effects associated with neurodegeneration in ALS. Aggregated SOD1G93A, released from murine NSC-34 motor neurons transiently transfected with SOD1G93A, could be transmitted to naïve NSC-34 cells and murine EOC13 microglia to induce endoplasmic reticulum (ER) stress and tumour necrosis factor-alpha (TNFα) release, respectively. Immunoblotting revealed NSC-34 cells expressed P2X7. Extracellular ATP induced cation dye uptake into these cells, which was blocked by the P2X7 antagonist AZ10606120, demonstrating these cells express functional P2X7. Moreover, ATP induced the rapid release of aggregated SOD1G93A from NSC-34 cells transiently transfected with SOD1G93A, a process blocked by AZ10606120 and revealing a role for P2X7 in this process. ATP-induced SOD1G93A release coincided with membrane blebbing. Finally, aggregated SOD1G93A released via P2X7 activation could also be transmitted to NSC-34 and EOC13 cells to induce ER stress and TNFα release, respectively. Collectively, these results identify a novel role for P2X7 in the prion-like propagation of SOD1 in ALS and provide a possible explanation for the therapeutic benefits of P2X7 antagonism previously observed in ALS SOD1G93A mice.

• Authors: Rachael Bartlett, Diane Ly, Neil R Cashman, Ronald Sluyter, Justin J Yerbury
• Journal: Purinergic signalling (Purinergic Signal) Vol. 18 Issue 4 Pg. 451-467 (12 2022) ISSN: 1573-9546 [Electronic] Netherlands
• PMID: 35478453 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2022. The Author(s).

Chemical References

• Adenosine Triphosphate
• AZ10606120
• Receptors, Purinergic P2X7
• Superoxide Dismutase-1
• Tumor Necrosis Factor-alpha
• P2rx7 protein, mouse

Topics

• Animals
• Mice
• Adenosine Triphosphate (pharmacology)
• Amyotrophic Lateral Sclerosis (pathology)
• Disease Models, Animal
• Mice, Transgenic
• Motor Neurons (metabolism)
• Receptors, Purinergic P2X7 (metabolism)
• Superoxide Dismutase-1 (metabolism)
• Tumor Necrosis Factor-alpha (metabolism)