Purinergic signaling plays a crucial role in somatosensory and nociceptive transmission under pathological conditions, such as spinal cord injury (SCI). Selective antagonists and allosteric modulators of P2X4 and P2X7 receptors are under investigation for neuropathic pain management. In our previous studies, we monitored endogenous real-time glutamate release during in vitro SCI experimental protocols induced by kainate (KA) (Mazzone and Nistri, Neurochem Int. 2019, 128:175-185). This study aimed to evaluate the role of purinergic signaling following chemical SCI in a mouse model. Our results revealed endogenous ATP release induced by KA (100 μM), monitored using a commercial biosensor, concomitantly with glutamate. P2X4 and P2X7 receptor expression was evaluated by RT-PCR after KA treatment. Additionally, we explored the effects of Coomassie Brilliant Blue G (10 μM, CBO), a non-nucleotide purinergic antagonist, on pyknotic cell death, as well as on the number of neurons, astrocytes, and microglia. Immunohistochemical analysis showed a significant reduction in neuronal loss in the KA+CBO group, with no observable effects on glial cells. Finally, in vivo administration of KA led to a significant impairment of locomotor function, which was partially reversed by CBO co-treatment. These findings suggest that early ATP release following excitotoxic injury may modulate neuronal survival and locomotor network function. Supported by Universidad Austral, CONICET, FONCYT, and IBRO.