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First published online 6 August 2008
doi: 10.1242/dev.024802

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V2a and V2b neurons are generated by the final divisions of pair-producing progenitors in the zebrafish spinal cord

National Institutes of Natural Sciences, Okazaki Institute for Integrative Bioscience, National Institute for Physiological Sciences, Higashiyama 5-1, Myodaiji, Okazaki, Aichi 444-8787, Japan.

View larger version (75K): [in this window] [in a new window]   Fig. 1. vsx1 mRNA and vsx1:GFP expression in spinal cord. (A,B) vsx1 mRNA expression in 22 hpf embryos. Broken lines outline the boundary of the spinal cord. (A) Cross-section. (B) Lateral view of a whole mount. (C) Tg[vsx1:GFP] fish at 2.5 dpf. (D,E) Expression of GFP and Vsx1 in Tg[vsx1:GFP] embryos at (D) 17 hpf and (E) 19 hpf. Arrows indicate pairs of labeled cells. Arrowheads indicate single labeled cells.

View larger version (73K): [in this window] [in a new window]   Fig. 2. Time-lapse imaging of vsx1-GFP cells showing that V2a and V2b neurons are produced as siblings. (A,B) Time-lapse imaging of vsx1-GFP cells, followed by immunohistochemical localization of Chx10 (red, V2a) and Scl (green, V2b). (A) Lateral view. (B) Dorsal view. Images are montages of stacked optical sections. Green arrowheads indicate cells prior to division. Red arrowheads indicate paired cells after division. Movies for these analyses are available in the supplementary material. (C) Summary of time-lapse imaging.

View larger version (42K): [in this window] [in a new window]   Fig. 3. Division axes of p2 intermediate progenitors are not fixed. (A,B) Confocal stacked images of vsx1-GFP sibling cells immediately after division. Embryos were analyzed at around 16-17 hpf. (A1-A4) Lateral views. Dorsal is towards the top. (B1-B4) Dorsal views. Superficial is towards the top. (C) Schematic of the method for division axis measurements. For each division, a line connecting the centers of the two sister cell (projection of division axis) was drawn. The angle of this line to the horizontal line (lateral views) or to the midline (dorsal views) was measured. (D,E) Quantification of the division angle.

View larger version (16K): [in this window] [in a new window]   Fig. 4. Model of V2 neuron development from a p2 intermediate progenitor. In this model of one unit of lineage, a single p2 intermediate progenitor produces two sibling cells. These cells then undergo Delta-Notch-mediated cell-cell interactions, resulting in different levels of Notch activity between them. The cell with higher Notch activity develops into a V2b neuron and the sister cell with lower activity (but higher levels of Delta expression) develops into a V2a neuron.

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