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. 2011 Jul 29;30(18):3830–3841. doi: 10.1038/emboj.2011.249

Figure 7.

Figure 7

MiR-103 dynamically regulates all Cav1.2-LTC subunits in vivo and is involved in pain sensitization. (A) In SNL animals, all three Cav1.2 subunit mRNAs were over-expressed as compared with sham (Cacna1c +138.57%, P<0.001; Cacna2d1 +47.09%, P<0.001; Cacnb1 +84.44%, P=0.005) while miR-103 was significantly decreased (−28.3%, P<0.001). ††P<0.01, †††P<0.001, ANOVA, Holm–Sidak post hoc test. MiR-103 intrathecal application significantly reduced Cacna1c, Cacna2d1 and Cacnb1 expressions (as compared with SNL level, −78.62%, P<0.001, −58.72%, P<0.001, −42.93%, P=0.004, respectively). In contrast, seed-mut-miR-103 did not change Cav1.2-LTC subunit expression. **P<0.01, ***P<0.001, ANOVA, Holm–Sidak post hoc test. Data are expressed as mean values±s.e.m. n=6, 6 and 3 for SNL, miR-103 and seed-mut-miR-103, respectively. (B) Rats were tested for pain sensitization using an electronic von Frey device: 7 days after surgery, they showed a decrease in paw withdrawal threshold in response to mechanical stimulation (40.70% of the pre-SNL threshold). Four daily intrathecal applications of miR-103 (days 7–10) alleviated mechanical allodynia (76.14% of pre-SNL threshold, P=0,031). Seed-mut-miR-103-injected animals showed no response to treatment (42.41% of pre-SNL threshold, P=0.219). *P<0.05, Wilcoxon signed rank test. Data are expressed as mean values±s.e.m. n=4, 4 and 6 for sham, miR-103 and seed-mut-miR-103-injected rats, respectively. (C) Cold allodynia was evaluated with a cold-plate device where animals are placed on a thermostatic steel plate set to +4°C. Elevation of the operated hind limb latency is measured and compared with the latency of the same animal just before surgery. Seven days after surgery (nerve ligation), SNL animals showed hypersensitivity to cold (23.53% of pre-surgery latency). Intrathecal miR-103 injections significantly reduced cold allodynia in SNL rats (58.75% of pre-surgery latency) whereas seed-mut-miR-103-injected animals showed no response to treatment. Data are expressed as mean values±s.e.m. n=6 and 4 for miR-103 and seed-mut-miR-103-injected rats, respectively.