Inhibition of spinal cytosolic phospholipase A 2 expression by an antisense oligonucleotide attenuates tissue injury-induced hyperalgesia

Abstract

Activation of the spinal phospholipase A 2 (PLA 2) –cyclooxygenase (COX) –prostaglandin signaling pathway is widely implicated in nociceptive processing. Although the role of spinal COX isoforms in pain signal transmission has been extensively characterized, our knowledge of PLA 2 enzymes in this cascade is limited. Among all PLA 2 groups, cytosolic calcium-dependent PLA 2 group IVA (cPLA 2IVA) appears to be the predominant PLA 2 enzyme in the spinal cord. In the present study we sought to (i) characterize anatomical and cellular distribution and localization of cPLA 2IVA in dorsal horn of rat spinal cord, (ii) verify efficacy and selectivity of intrathecal (IT) delivery of an antisense oligonucleotide (AS) targeting rat cPLA 2IVA mRNA on spinal expression of this enzyme, and (iii) examine the effect of down-regulation of spinal cPLA 2IVA on peripheral tissue injury–induced pain behavior. Here we demonstrate that cPLA 2IVA is constitutively expressed in rat spinal cord, predominantly in dorsal horn neurons and oligodendrocytes but not in astrocytes or microglia. Intrathecal injection of AS significantly down-regulated both protein and gene expression of cPLA 2IVA in rat spinal cord, while control missense oligonucleotide (MS) had no effect. Immunocytochemistry confirmed that the reduction occurred in neurons and oligodendrocytes. cPLA 2IVA AS did not alter expression of several other PLA 2 isoforms, such as secretory PLA 2 (groups IIA and V) and calcium-independent PLA 2 (group VI), indicating that the AS was specific for cPLA 2IVA. This selective knockdown of spinal cPLA 2IVA did not change acute nociception (i.e. paw withdrawal thresholds to acute thermal stimuli and intradermal formalin-induced first phase flinching), however, it significantly attenuated formalin-induced hyperalgesia (i.e. second phase flinching behavior), which reflects spinal sensitization. Thus the present findings suggest that cPLA 2IVA may specifically participate in spinal nociceptive processing.