Figure 2

Inhibition of Aurora Kinases A, B, A/B, B/C, and A/B/C in primary cortical neurons suggests roles for Aurora B and C in neurite initiation/elongation and arborization. (A) Primary cortical mouse neurons transfected with YFP and treated with Aurora kinase inhibitors, scale bars 50 µm. (B) One-way ANOVA showed significant difference in length of neurites likely to become dendrites [F(5,354) = 16.941, p < 0.0005]. There was a significant decrease in neurons treated with Aurora B inhibitor (10.6 ± 0.95, n = 64, p < 0.0005), Aurora B/C inhibitor (7.7 ± 0.86, n = 46, p < 0.0005), or Aurora A/B/C inhibitor (15.5 ± 1.21, n = 39, p = 0.49) compared to DMSO control (21.0 ± 1.29, n = 80). There was no significant difference between neurons treated with Aurora A inhibitor (18.9 ± 1.26, n = 74, p = 1.000) or Aurora A/B inhibitor (17.4 ± 1.38, n = 57, p = 0.437) compared to DMSO control. (C) One-way ANOVA showed significant difference in length of neurites likely to become axons [F(5,111) = 12.946, p < 0.0005]. There was a significant decrease in neurons treated Aurora B inhibitor (25.7 ± 4.08, n = 20, p < 0.0005), Aurora B/C inhibitor (19.6 ± 1.36, n = 20, p < 0.0005), or Aurora A/B/C inhibitor (23.4 ± 1.73, n = 19, p < 0.0005) compared to DMSO treated control (54.1 ± 5.33, n = 20). There was no significant difference between neurons treated with Aurora A inhibitor (43.8 ± 5.13, n = 20, p = 0.860) or Aurora A/B inhibitor (40.7 ± 2.98, n = 18, p = 0.247) compared to DMSO control. (D) One-way ANOVA showed significant difference in mean number of neurites [F(5,113) = 8.941, p < 0.0005]. There was a significant decrease in neurons treated with Aurora A/B inhibitor (3.68 ± 0.47, n = 19, p = 0.017), Aurora B/C inhibitor (3.25 ± 0.32, p = 0.001), and Aurora A/B/C inhibitor (2.95 ± 0.28, p < 0.0005) compared to the DMSO treated controls (5.60 ± 0.39). There was no significant difference between neurons treated with Aurora A inhibitor (5.50 ± 0.50, p = 1.000) or Aurora B inhibitor (5.20 ± 0.43, p = 1.000) and DMSO control. For all groups, n = 20 unless otherwise noted. (E) Sholl analysis shows dramatic decrease in branching after treatment with Aurora Kinase B, A/B, B/C, and A/B/C inhibitors and a mild decrease in branching after Aurora Kinase A inhibitor treatment.