Objectives. We aim to determine the effects of Botulinum toxin type A (BTX-A) on neuropathic pain behavior and the expression of P2X3 receptor in dorsal root ganglion (DRG) in rats with neuropathic pain induced by L5 ventral root transection (L5 VRT). Methods. Neuropathic pain was induced by L5 VRT in male Sprague-Dawley rats. Either saline or BTX-A was administered to the plantar surface. Behavioral tests were conducted preoperatively and at predefined postoperative days. The expression of P2X3 receptors in DRG neurons was detected by immunoreactivity at postoperative days 3, 7, 14, and 21. Results. The number of positive P2X3 neurons in the ipsilateral L5 DRG increased significantly after L5 VRT ( P < 0.001). This increase persisted for at least 3 weeks after the operation. No significant changes in P2X3 expression were detected in the contralateral L5, or in the L4 DRGs bilaterally. Subcutaneous administration of BTX-A, performed on the left hindpaw at days 4, 8, or 16 post VRT surgery, significantly reduced mechanical allodynia bilaterally and inhibited P2X3 over-expression induced by L5 VRT. Conclusions. L5 VRT led to over-expression of P2X3 receptors in the L5 DRG and bilateral mechanical allodynia in rats. Subcutaneous injection of BTX-A significantly reversed the neuropathic pain behavior and the over-expression of P2X3 receptor in nociceptive neurons. These data not only show over-expression of purinergic receptors in the VRT model of neuropathic pain but also reveal a novel mechanism of botulinum toxin action on nociceptive neurons. [ABSTRACT FROM AUTHOR]