Figure 6

Laser ablation of the centriole in larval ddaE neurons does not affect microtubule polarity. (A) Bleaching of the centriole yields recovery of RFP-Fzr within 2 days. The red circle indicates the area containing the centriole that was bleached. Immediately post-bleach, no concentrated RFP-Fzr spot is seen compared to 24 hours post-bleach, at which time recovery has occurred (circle). As shown by the chart, recovery typically occurs within 24 hours (11 of 12 cells, and the remaining cell recovered by 48 hours). (B) Without laser ablation, the centriole persists in neurons. RFP-Fzr was imaged in ddaE neurons over a 72 hour time period. Six neurons from 6 larvae were imaged. Unlike centriole-ablated cells (Figure 6D), RFP-Fzr is clearly seen in the cell over all 3 imaging days. (C) Centriole ablation has no effect on the directionality of growing microtubules, which was quantified in class 1 ddaE neurons 1, 2, and 3 days after centriole ablation. The 'n' values are the number of EB1-GFP comets analyzed. Six neurons from 6 larvae were examined for both conditions. In all cases, P > 0.05 (Fisher's exact test), which indicates that the neurons retain their polarity despite ablation of the centriole. (D) Laser ablation eliminates RFP-Fzr concentrations. In this example, pre-ablation, the centriole is clearly seen in the cell body (circle). Post-ablation, the centriole appears to have fragmented. This is more clearly seen 24 hours post-ablation; past 24 hours, there are no noticeable concentrations of RFP-Fzr.