Figure 5.
Figure 5. RGCs can switch to a multipolar migratory mode. (A) Rare example of multipolar migration in control embryos. Bar, 5 µm. (B) Multipolar migration induced by MT destabilization. Stathmin 1 overexpression induced at 30 hpf. Time lapse starts at 34 hpf. (A and B) Gray phase, cell still has the basal and apical process (A) or apical process (B); green phase, directional multipolar mode; white dots, RGC followed; arrowheads, apical and basal process (A) or apical process (B); asterisks, apical process loss; blue arrows, axon. Bar, 10 µm. (C) Typical trajectory of RGCs with destabilized MTs from the montage in B. Arrowheads, loss of the apical process (AP). More trajectories are in Fig. S4 E. The mean wild-type (WT) trajectory is shown. (D) MSDs of RGCs in multipolar migratory mode. Values are taken from the first 95 min after apical process loss. Wild type from Fig. 1 E and RGCs without basal process from Fig. 2 G. The α value is given with a 95% confidence interval. (E) Directionality ratios before and after apical process loss. Values are taken from the first 95 min after mitosis and the first 95 min after apical process loss. Final directionality ratios: after apical process (AP) loss, 0.71; after mitosis, 0.45. Wild type from Fig. 1 F and RGCs without basal process (BP) from Fig. 2 J. (F) Multipolar migration induced by Arp2/3 inhibition. ath5:gap-RFP fish were imaged in a spinning disk microscope from 34 hpf. CK-666 was added at the start of imaging. Images were denoised in Fiji (ROF denoise). (G) Multipolar migration induced by Arp2/3 inhibition. NWASP-CA overexpression was induced at 30 hpf. Time lapse starts at 34 hpf. (F and G) Gray phase, cell has apical process; green phase, directional multipolar mode; white dots, RGC followed; arrowheads, apical process; asterisks, apical process loss. Dashed lines delimit the apical and basal sides. Bars, 10 µm. (H) Typical trajectory of RGCs upon NWASP-CA overexpression (OE) from the montage in G. Arrowhead, apical process loss. More trajectories are in Fig. S4 H. The mean wild-type trajectory is shown. (I) MSDs of RGCs after Arp2/3 inhibition (NWASP-CA overexpression). Wild type from Fig. 1 E and RGCs without basal process from Fig. 2 G. The α value is given with a 95% confidence interval. (J) Directionality ratios of RGCs after mitosis and after apical process loss upon NWASP-CA overexpression. (I and J) Values are taken from the first 95 min after mitosis and the first 95 min after apical process loss. Error bars represent SEM. Final directionality ratios: after apical process loss = 0.63; after mitosis = 0.70. Wild type from Fig. 1 F and RGCs without BP from Fig. 2 J.

RGCs can switch to a multipolar migratory mode. (A) Rare example of multipolar migration in control embryos. Bar, 5 µm. (B) Multipolar migration induced by MT destabilization. Stathmin 1 overexpression induced at 30 hpf. Time lapse starts at 34 hpf. (A and B) Gray phase, cell still has the basal and apical process (A) or apical process (B); green phase, directional multipolar mode; white dots, RGC followed; arrowheads, apical and basal process (A) or apical process (B); asterisks, apical process loss; blue arrows, axon. Bar, 10 µm. (C) Typical trajectory of RGCs with destabilized MTs from the montage in B. Arrowheads, loss of the apical process (AP). More trajectories are in Fig. S4 E. The mean wild-type (WT) trajectory is shown. (D) MSDs of RGCs in multipolar migratory mode. Values are taken from the first 95 min after apical process loss. Wild type from Fig. 1 E and RGCs without basal process from Fig. 2 G. The α value is given with a 95% confidence interval. (E) Directionality ratios before and after apical process loss. Values are taken from the first 95 min after mitosis and the first 95 min after apical process loss. Final directionality ratios: after apical process (AP) loss, 0.71; after mitosis, 0.45. Wild type from Fig. 1 F and RGCs without basal process (BP) from Fig. 2 J. (F) Multipolar migration induced by Arp2/3 inhibition. ath5:gap-RFP fish were imaged in a spinning disk microscope from 34 hpf. CK-666 was added at the start of imaging. Images were denoised in Fiji (ROF denoise). (G) Multipolar migration induced by Arp2/3 inhibition. NWASP-CA overexpression was induced at 30 hpf. Time lapse starts at 34 hpf. (F and G) Gray phase, cell has apical process; green phase, directional multipolar mode; white dots, RGC followed; arrowheads, apical process; asterisks, apical process loss. Dashed lines delimit the apical and basal sides. Bars, 10 µm. (H) Typical trajectory of RGCs upon NWASP-CA overexpression (OE) from the montage in G. Arrowhead, apical process loss. More trajectories are in Fig. S4 H. The mean wild-type trajectory is shown. (I) MSDs of RGCs after Arp2/3 inhibition (NWASP-CA overexpression). Wild type from Fig. 1 E and RGCs without basal process from Fig. 2 G. The α value is given with a 95% confidence interval. (J) Directionality ratios of RGCs after mitosis and after apical process loss upon NWASP-CA overexpression. (I and J) Values are taken from the first 95 min after mitosis and the first 95 min after apical process loss. Error bars represent SEM. Final directionality ratios: after apical process loss = 0.63; after mitosis = 0.70. Wild type from Fig. 1 F and RGCs without BP from Fig. 2 J.

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