FIGURE

Fig. 3

ID

ZDB-FIG-170908-10

Publication

Breau et al., 2017 - Extrinsic mechanical forces mediate retrograde axon extension in a developing neuronal circuit

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Fig. 3

Axonal identity of the cytoplasmic protrusions. All images represent dorsal views; dotted white lines indicate the brain surface. a, b XY sections extracted from a movie performed on a ngn1:gfp embryo injected with H2B-RFP mRNA. From 18 s onwards, the cytoplasmic processes connecting cell bodies to the brain form a bundle of GFP+ protrusions juxtaposed to the brain surface, in the ventromedial region of the placode (framed with white lines). c Live imaging on a wild-type embryo transplantated with Doublecortin–GFP-expressing cells, showing microtubules in the shaft of the protrusions and around cell bodies during cell movements. d Acetylated tubulin immunostaining (magenta) performed on a ngn1:gfp embryo, indicating the presence of stable microtubules in GFP+ protrusions and cell bodies at 24 s. e Mosaic labelling of actin and membranes, obtained with a transplantation of Utrophin-GFP (green) and mbCherry (magenta) expressing cells in a wild-type embryo. f Immunostaining for the OP pioneer axon marker Zns2 (magenta) on a ngn1:gfp embryo, labelling the bundle of GFP+ protrusions at 26 hpf (white lines). g High magnification of OP cells expressing mbCherry and the axonal specification marker Kif5c560-YFP, showing the accumulation of Kif5c560-YFP at the tip of the protrusions (arrowhead) during lateral movements. On the left, the embryo was co-injected with Kif5c560-YFP mRNA and pCS2-mbCherry DNA, explaining why some of the Kif5c560-YFP + accumulations are not associated with magenta cells. On the right, a wild-type embryo was transplanted with cells co-expressing Kif5c560-YFP and mbCherry mRNAs. h Long-term live imaging of an embryo injected with mbCherry (green) mRNA, from 18 s to 26 hpf stages. A few OP cells express higher levels of mbCherry, which allows to visualise the elongation of their protrusions during lateral movements (white arrowheads), and their entry into the brain territory (green arrowhead). i Zns2 immunostaining (magenta) performed on the embryo imaged in h, showing the mbCherry+ protrusions within the Zns2+ bundle at 26 hpf. Scale bars: 25 µm in a, c, d, f, h, i, and 10 µm in e, g

Expression Data

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Antibody Labeling

Phenotype Data

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Acknowledgments

This image is the copyrighted work of the attributed author or publisher, and ZFIN has permission only to display this image to its users. Additional permissions should be obtained from the applicable author or publisher of the image. Full text @ Nat. Commun.