The damage-associated molecular pattern receptor P2X7 is an ATP-gated cation channel, which plays an important role in neuroinflammation. P2X7 has been implicated in a number of neurodegenerative diseases, including amyotrophic lateral sclerosis (ALS). In ALS, the extracellular release of mutant superoxide dismutase 1 (mSOD1) from motor neurons is a feature thought to contribute to disease progression. Thus, this study aimed to determine if P2X7 activation mediates the release of ALS-associated mSOD1 from NSC-34 murine motor neuron-like cells transiently transfected with mSOD1:GFP. Immunoblotting confirmed the presence of P2X7 protein in NSC-34 cells. Flow cytometric measurements of cation uptake demonstrated that these cells express functional P2X7. Immunoblotting revealed that the P2X7 agonist ATP induced significant mSOD1 release from NSC-34 cells. This ATP-induced mSOD1 release was completely inhibited by pre-treatment with the P2X7 antagonist AZ10606120. Collectively, these results indicate that P2X7 is capable of mediating the extracellular release of mSOD1 from motor neurons. Thus, P2X7 may represent a therapeutic target in ALS. The mechanism of P2X7 mediated SOD1 release from motor neurons and the effect of cations on this process is currently under investigation.