Figure 7.
Imbalanced cargo transport reduces THN motility.(A) Schematic representation of Kif1C-5C chimera. (B and C) In THNs, the chimera localizes to the front of phase 1 (B) and phase 2 (C) cells. Cyan box indicates magnified region on the right. Scale bars represent 20 µm in overview images and 5 µm in magnified images. (D) Kif1C-5C does not colocalize with Cadherin-2 particles, although they are frequently found in the vicinity. Colored boxes indicate magnified regions on the right. Scale bars represent 10 µm in the overview image and 1 µm in magnified images. (E) The expression of Kif1C-5C increases stationary times of Cadherin-2 particles. For full statistical testing, see Table S2; n = 416 tracks for injection control, n = 308 tracks for p50-expressing embryos, n = 593 tracks for Kif5C-expressing embryos, n = 441 tracks Kif5C T94N-expressing embryos, and n = 439 tracks for Kif1C-5C–expressing embryos. (F and G) Kif1C-5C also reduces maximal velocities and mean velocities (F; area of 0.8 data density in top left corner and number of tracks in lower left corner; see also Video 6) as well as instantaneous velocities (G). Note that the retrograde direction is more strongly affected than the anterograde direction, as evidenced by the comparison to other motor-affecting treatments. For full statistical testing, see Table S3 and Table S4. Anterograde: n = 2,059 values for injection control, n = 1,044 values for p50-expressing embryos, n = 2,808 values for Kif5C-expressing embryos, n = 2,055 values Kif5C T94N-expressing embryos, and n = 2,245 values for Kif1C-5C–expressing embryos. Retrograde: n = 2,127 values for injection control, n = 1,117 values for p50-expressing embryos, n = 2,791 values for Kif5C-expressing embryos, n = 2,132 values Kif5C T94N-expressing embryos, and n = 2,187 values for Kif1C-5C–expressing embryos. (H and I) Kif1C-5C reduces overall THN motility. Elapsed time in minutes. Scale bars, 20 µm. n = 24 embryos/145 tracks for control and n = 20 embryos/142 tracks for Kif1C-5C–expressing embryos. Kruskal–Wallis ANOVA: P = 0.007. See also Video 4. (J) Models of how MTs and motors contribute to neuronal migration, with nucleokinesis on the left and the spatiotemporal distribution model on the right. MHB, solid line; URL, dotted line. Boxes in graphs represent 25–75% of all values and whiskers 1.5 times the quartile. Median is shown as a horizontal bar and mean as a square box. Significance level in Kruskal–Wallis ANOVA: *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant. All data shown in E–G represent n = 13 embryos for injection control, n = 14 p50-expressing embryos, n = 11 Kif5C-expressing embryos, n = 16 Kif5C T94N-expressing embryos, and n = 9 Kif5C-1C–expressing embryos.

Imbalanced cargo transport reduces THN motility. (A) Schematic representation of Kif1C-5C chimera. (B and C) In THNs, the chimera localizes to the front of phase 1 (B) and phase 2 (C) cells. Cyan box indicates magnified region on the right. Scale bars represent 20 µm in overview images and 5 µm in magnified images. (D) Kif1C-5C does not colocalize with Cadherin-2 particles, although they are frequently found in the vicinity. Colored boxes indicate magnified regions on the right. Scale bars represent 10 µm in the overview image and 1 µm in magnified images. (E) The expression of Kif1C-5C increases stationary times of Cadherin-2 particles. For full statistical testing, see Table S2; n = 416 tracks for injection control, n = 308 tracks for p50-expressing embryos, n = 593 tracks for Kif5C-expressing embryos, n = 441 tracks Kif5C T94N-expressing embryos, and n = 439 tracks for Kif1C-5C–expressing embryos. (F and G) Kif1C-5C also reduces maximal velocities and mean velocities (F; area of 0.8 data density in top left corner and number of tracks in lower left corner; see also Video 6) as well as instantaneous velocities (G). Note that the retrograde direction is more strongly affected than the anterograde direction, as evidenced by the comparison to other motor-affecting treatments. For full statistical testing, see Table S3 and Table S4. Anterograde: n = 2,059 values for injection control, n = 1,044 values for p50-expressing embryos, n = 2,808 values for Kif5C-expressing embryos, n = 2,055 values Kif5C T94N-expressing embryos, and n = 2,245 values for Kif1C-5C–expressing embryos. Retrograde: n = 2,127 values for injection control, n = 1,117 values for p50-expressing embryos, n = 2,791 values for Kif5C-expressing embryos, n = 2,132 values Kif5C T94N-expressing embryos, and n = 2,187 values for Kif1C-5C–expressing embryos. (H and I) Kif1C-5C reduces overall THN motility. Elapsed time in minutes. Scale bars, 20 µm. n = 24 embryos/145 tracks for control and n = 20 embryos/142 tracks for Kif1C-5C–expressing embryos. Kruskal–Wallis ANOVA: P = 0.007. See also Video 4. (J) Models of how MTs and motors contribute to neuronal migration, with nucleokinesis on the left and the spatiotemporal distribution model on the right. MHB, solid line; URL, dotted line. Boxes in graphs represent 25–75% of all values and whiskers 1.5 times the quartile. Median is shown as a horizontal bar and mean as a square box. Significance level in Kruskal–Wallis ANOVA: *, P < 0.05; **, P < 0.01; ***, P < 0.001; ns, not significant. All data shown in E–G represent n = 13 embryos for injection control, n = 14 p50-expressing embryos, n = 11 Kif5C-expressing embryos, n = 16 Kif5C T94N-expressing embryos, and n = 9 Kif5C-1C–expressing embryos.

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