Research Paper Volume 13, Issue 1 pp 389—410

DOK3 is involved in microglial cell activation in neuropathic pain by interacting with GPR84

GPR84 activation-induced mechanical allodynia and inflammatory responses are compromised in DOK3-/- mice. B129 and DOK3-/- mice were treated with 6-OAU (1 μM) for 0, 1, 2, and 4 h via intrathecal injection. (A) GPR84 activation-induced mechanical allodynia was determined by calculating PWMT at 0, 1, 2, and 4 h after intrathecal injection. N=8-10, data are presented as means ± SEM. *p  0.05 vs. control (CON) or 0 h; & p  0.05 vs. WT mice at the same time after intrathecal injection. (B) DOK3 mRNA levels in mouse lumbar spinal cord were determined by RT-qPCR. (C) DOK3 protein levels were determined by western blotting 2 h after 6-OAU injection (1 μM). (D–F) Lumbar spinal cords were harvested from mice to determine the levels of mRNAs for TNF-α (D), IL-6 (E), and IL-1β (F) 2 h after 6-OAU injection (1 μM). (G) B129 mice were treated with 6-OAU (1 μM) for 2 h. The co-localization of GPR84 and DOK3 proteins was assessed using immunofluorescence analysis in mice. Scale bar, 20 μm. (H) The expression of CD11B proteins in lumbar spinal cord was assayed by immunofluorescence analysis and quantified (I). N=8-10, data are presented as means ± SEM. *p  0.05, #p  0.01 for comparisons between the 2 groups connected by the horizontal line; NS, not significant; scale bar, 50 μm.

Figure 5. GPR84 activation-induced mechanical allodynia and inflammatory responses are compromised in DOK3-/- mice. B129 and DOK3-/- mice were treated with 6-OAU (1 μM) for 0, 1, 2, and 4 h via intrathecal injection. (A) GPR84 activation-induced mechanical allodynia was determined by calculating PWMT at 0, 1, 2, and 4 h after intrathecal injection. N=8-10, data are presented as means ± SEM. *p < 0.05 vs. control (CON) or 0 h; & p < 0.05 vs. WT mice at the same time after intrathecal injection. (B) DOK3 mRNA levels in mouse lumbar spinal cord were determined by RT-qPCR. (C) DOK3 protein levels were determined by western blotting 2 h after 6-OAU injection (1 μM). (DF) Lumbar spinal cords were harvested from mice to determine the levels of mRNAs for TNF-α (D), IL-6 (E), and IL-1β (F) 2 h after 6-OAU injection (1 μM). (G) B129 mice were treated with 6-OAU (1 μM) for 2 h. The co-localization of GPR84 and DOK3 proteins was assessed using immunofluorescence analysis in mice. Scale bar, 20 μm. (H) The expression of CD11B proteins in lumbar spinal cord was assayed by immunofluorescence analysis and quantified (I). N=8-10, data are presented as means ± SEM. *p < 0.05, #p < 0.01 for comparisons between the 2 groups connected by the horizontal line; NS, not significant; scale bar, 50 μm.