Figure 1.
Figure 1. EB proteins label two types of puncta in Drosophila and zebrafish. (A) Whole animal image of a 3-d-old larva labeled with EB1-GFP under 221-Gal4. Arrows point to class I ddaE neurons. (B and C) Quantification of direction and speed of two types of puncta labeled by EB1-GFP in Drosophila ddaE comb dendrites. The speed cutoff between fast and slow puncta was 1.6 µm/min. Numbers on the graphs are total numbers of dots recorded from 16 neurons for fast dots and 35 for slow dots. One neuron was used per animal throughout this study. Error bars are SD for all the graphs presented. Turquoise is used to show fast dots and pink to indicate slow ones in all figures. (D) Representative image of a class I ddaE neuron expressing EB1-GFP from a 3-d-old larva. Cell body and dendrites were shown with axon extending to lower left corner toward ventral nerve cord. Dashed line indicates the comb dendrite where observations of EB1-GFP puncta were performed. (E and F) Representative image and kymograph of a dendrite segment of a class I ddaE neuron expressing EB1-GFP from a time-lapse recording. Sequential images of the rectangular region are zoomed in to show a slow puncta (pink arrowhead) and a fast one (turquoise arrowhead). Horizontal bars, 5 μm. Vertical bars, 60 s. In all kymographs presented in this study, time flows from top to bottom, and the cell body is on the right side. (G) Representative kymograph of two fast dots initiating from a slow-moving one. Horizontal bar, 5 μm. Vertical bar, 60 s. (H) Representative image of an axon segment of class IV ddaC neuron expressing EB1-GFP from a time-lapse recording. Sequential images of the rectangular region are zoomed in to focus on a slow dot marked by a pink arrowhead. Multiple fast dots that moved across the same region were not marked. Horizontal bars, 5 μm. (I) Kymograph of EB1-GFP in the axon of the ddaC neuron. As in other images, the horizontal bar is 5 μm and vertical is 60 s. Turquoise lines show fast puncta, and pink, slow. The cell body is off the right side of the image. (J) Whole animal image of a 48-hpf zebrafish expressing EB3-GFP, tdTomato, and mCherry in RB neurons. EB3-GFP, mCherry, and tdTomato were controlled by the P2X3 and ISL1 promoters. (K) Zoomed-in view of RB neurons innervating the skin from the same animal imaged in H. (L and M) Representative image and kymograph of sensory endings of an RB neuron expressing EB3-GFP. The dashed line marks the region from which the kymograph was generated. (N and O) Quantification of direction and speed of puncta labeled by EB3-GFP in RB neurons of zebrafish. The speed cutoff between fast and slow puncta was 1.2 µm/min. Numbers on the graph are numbers of dots recorded from 6 neurons for fast puncta and 12 for slow puncta.

EB proteins label two types of puncta in Drosophila and zebrafish. (A) Whole animal image of a 3-d-old larva labeled with EB1-GFP under 221-Gal4. Arrows point to class I ddaE neurons. (B and C) Quantification of direction and speed of two types of puncta labeled by EB1-GFP in Drosophila ddaE comb dendrites. The speed cutoff between fast and slow puncta was 1.6 µm/min. Numbers on the graphs are total numbers of dots recorded from 16 neurons for fast dots and 35 for slow dots. One neuron was used per animal throughout this study. Error bars are SD for all the graphs presented. Turquoise is used to show fast dots and pink to indicate slow ones in all figures. (D) Representative image of a class I ddaE neuron expressing EB1-GFP from a 3-d-old larva. Cell body and dendrites were shown with axon extending to lower left corner toward ventral nerve cord. Dashed line indicates the comb dendrite where observations of EB1-GFP puncta were performed. (E and F) Representative image and kymograph of a dendrite segment of a class I ddaE neuron expressing EB1-GFP from a time-lapse recording. Sequential images of the rectangular region are zoomed in to show a slow puncta (pink arrowhead) and a fast one (turquoise arrowhead). Horizontal bars, 5 μm. Vertical bars, 60 s. In all kymographs presented in this study, time flows from top to bottom, and the cell body is on the right side. (G) Representative kymograph of two fast dots initiating from a slow-moving one. Horizontal bar, 5 μm. Vertical bar, 60 s. (H) Representative image of an axon segment of class IV ddaC neuron expressing EB1-GFP from a time-lapse recording. Sequential images of the rectangular region are zoomed in to focus on a slow dot marked by a pink arrowhead. Multiple fast dots that moved across the same region were not marked. Horizontal bars, 5 μm. (I) Kymograph of EB1-GFP in the axon of the ddaC neuron. As in other images, the horizontal bar is 5 μm and vertical is 60 s. Turquoise lines show fast puncta, and pink, slow. The cell body is off the right side of the image. (J) Whole animal image of a 48-hpf zebrafish expressing EB3-GFP, tdTomato, and mCherry in RB neurons. EB3-GFP, mCherry, and tdTomato were controlled by the P2X3 and ISL1 promoters. (K) Zoomed-in view of RB neurons innervating the skin from the same animal imaged in H. (L and M) Representative image and kymograph of sensory endings of an RB neuron expressing EB3-GFP. The dashed line marks the region from which the kymograph was generated. (N and O) Quantification of direction and speed of puncta labeled by EB3-GFP in RB neurons of zebrafish. The speed cutoff between fast and slow puncta was 1.2 µm/min. Numbers on the graph are numbers of dots recorded from 6 neurons for fast puncta and 12 for slow puncta.

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