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. 2014 Jul 16;34(29):9476–9483. doi: 10.1523/JNEUROSCI.5346-13.2014

Figure 3.

Figure 3.

miR-219 targeting CaMKIIγ regulates chronic inflammation pain. A, Validation of miR-219 targeting CaMKIIγ. A mutation was generated in the CaMKIIγ-3′-UTR sequence in the complementary site for the seed region of miR-219 as indicated (pCHK-mut-CaMKIIγ). **p < 0.01, versus pCHK-mut-CaMKIIγ or empty vector group. n = 6. B, C, The increased spinal CaMKIIγ expression in CFA mice was reversed by intrathecal injection of miR-219 mimics or Lenti-miR-219. **p < 0.01, versus adjacent groups. n = 5. CaMKIIγ was measured at 4 d after miR-219 mimic injection (6 d after CFA injection) or at 5 d after Lenti-miR-219 injection (7 d after CFA injection), respectively. D, Spinal CaMKIIγ expression was increased by intrathecal injection of miR-219-sponge in naive mice. **p < 0.01, versus adjacent groups. n = 5. CaMKIIγ was measured at 5 d after injection of miR-219-sponge. E, FISH (miR-219, green) immunofluorescent (CaMKIIγ, red) costaining in spinal cord at 3 d after CFA injection. Scale bar, 25 μm. F, G, Knockdown of CaMKIIγ significantly inhibited or reversed thermal hyperalgesia and mechanical allodynia induced by miR-219-sponge in naive mice. *p < 0.05, versus CaMKIIγ siRNA + miR-219-sponge or miR-219-sponge + CaMKIIγ siRNA group. n = 8. Blue arrow indicates CaMKIIγ siRNA or control siRNA injection; black arrow indicates miR-219-sponge injection. H, The increased expression of spinal p-NR1 in CFA-injected mice (left) or in miR-219-sponge-injected naive mice (right) was reversed by intrathecal injection of CaMKIIγ siRNA. **p < 0.01, versus adjacent groups. *p < 0.05, versus adjacent groups. n = 5. p-NR1 was measured at 4 d after CaMKIIγ or control siRNA injection (6 d after CFA injection), or at 5 d after CaMKIIγ or control siRNA injection (7 d after miR-219-sponge injection). I, MK-801 markedly inhibited thermal hyperalgesia and mechanical allodynia induced by miR-219-sponge in naive mice. *p < 0.05, versus miR-219-sponge + saline group. n = 8. MK-801 (10 nmol) was intrathecally injected at 30 min after pain behavior test. Blue arrow indicates MK-801 or saline injection; black arrow indicates miR-219-sponge injection.