Improvements in chemotherapy have increased the cancer patient's lifespan, but these drugs produce severe side-effects, including a distal-symmetrical sensory neuropathy that is often accompanied by a neuropathic pain syndrome. Chemotherapeutics in the taxane alkaloid classes such as Paclitaxel are among the most effective drugs for the treatment of solid tumors, but are also the most often associated with these neuropathies. The cause of the pain is not known. Changes in firing rates of primary afferents have been associated with chemotherapeutic agent-induced pain, and since glutamate is the principal excitatory neurotransmitter at the first central synapse of the sensory pain pathway, this study investigated whether changes in glutamate homeostasis occur within the spinal cord after Paclitaxel treatment. The novel method of in vivo glutamate voltammetry was utilized to determine if changes in spinal extracellular glutamate levels and release in mice occur after chronic Paclitaxel therapy. The study revealed that 4 –fold enhanced glutamate evoked glutamate release occurs on Day 8 after the initiation of Paclitaxel treatment, compared to vehicle-treated animals. Paclitaxel-induced tactile hyperalgesia was also well-established by Day 8 of treatment. This study suggests that activation of glutamate receptors stimulates augmented glutamate release in Paclitaxel-treated animals, and the resultant elevated extracellular glutamate activating spinal glutamate receptors comprises a new putative mechanism for the iatrogenically induced neuropathic pain of the taxane class of chemotherapeutic agents.