DEV02574 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure 1 -

  Fig. S1. Morphology of cervical sympathetic ganglia in 2 dpf zebrafish embryos. After whole-mount in situ hybridisation for th as a marker for sympathetic neurons, dorsal hindbrain/spinal cord and yolk sac were dissected and viewed from dorsal using 40× objective. Please note individual cells with nuclei spared from the in situ staining reaction in the plane of focus and other cells that give a blurred appearance, as they are out of focus. The embryo shown was selected for the number of cells within one focus plane. Somite numbers are indicated to document that cervical sympathetic ganglion cells are mainly located betwen somites 1 and 4.

• Supplemental Figure 2 -

  Fig. S2. Localisation of cervical sympathetic and enteric neurons in 3 dpf zebrafish embryos. (A-D) Whole-mount in situ hybridisation for phox2b as marker for autonomic cells was carried out. After dissection of dorsal hindbrain/spinal cord and yolk sac, a stack of pictures was made focussing from dorsal (A) to ventral (D) through the embryo. This illustrates the dorsal location of the cervical sympathetic ganglion (dashed circle), restricted to a relatively short anteroposterior area (A). Enteric neurons (white arrowheads) that appear as diffuse staining in A show up in more ventral focal planes (B-D), and extend to more rostral and caudal regions down to the anus (not shown). (E) An optical overlay of images in A to D to illustrate the different lateral positions of enteric and cervical sympathetic ganglion cells.

• Supplemental Figure 3 -

  Fig. S3. Rescue of th expression in cervical sympathetic ganglia of 2 dpf hands off zebrafish embryos. hands off mutant embryos, (A) without or (B) with injection of synthetic hand2 mRNA, stained for th expression. Rescued hands off mutants displayed a highly significant increase in th-positive cells when compared with uninjected hands off embryos (see Fig. 2N)