Figure 2

Intrathecal injection of AAV infects lumbar DRG neurons. Following intrathecal injections of AAV2/5 vectors (with GFP reporters). (A) DRG were stained with GFP and NeuN. A merged image in the top panel in A shows that cell bodies immunopositive for GFP (cells infected by vectors) colocalize with NeuN, as shown by yellow color. (B) A similar histological approach in spinal cord tissue from the same animals revealed no colocalization of GFP and NeuN, demonstrating that AAV vectors did not infect spinal cord neurons (see merged bottom panel in B). As expected, some GFP-labeled afferent fibers from infected DRGs appeared in the dorsal column white matter tracts and in the deeper laminae of the dorsal horn gray matter (white arrowheads). (C) Normalized Nav1.3 quantitative PCR from lumbar enlargement DRGs 3 wks after AAV intrathecal injection demonstrate significant differences in Nav1.3 mRNA expression following shRNA-Nav1.3 compared with scrambled AAV vector treatment.