XB-IMG-170212
Xenbase Image ID: 170212
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Figure 2. Zebrafish NECs are not neural crest-derived: analysis of neural crest-deficient zebrafish mutants.(a) 7-dpf tfap2amob;foxd3mos zebrafish lack all neural crest derivatives, including melanophores and jaw skeleton (arrowhead). Dotted lines: section planes in b-e. (b,c) At 7-dpf, tfap2amob;foxd3mos orobranchial epithelium retains innervated serotonergic (5-HT+) cells (putative NECs), identified by cytoplasmic serotonin surrounded by a ring of HNK1 epitope-immunoreactive neurites. (d,e) At 7-dpf, putative NECs (arrowheads) persist in tfap2amob;foxd3mos zebrafish in the region ventral to the orobranchial cavity where the pharyngeal arches would be located in wild-type fish. (NB The hypothalamus [Hyp] extends caudally beneath the midbrain and rostral hindbrain, and often separates from the overlying brain on sections, as here.) (f) The mean number per 7-dpf larva of putative NECs in the orobranchial epithelium does not differ between tfap2amob;foxd3mos (98.0 ± 49.7 s.d.; n = 3) and wild-type zebrafish (81.3 ± 24.0 s.d.; n = 3) (p=0.63, unpaired two-tailed Student’s t-test). All such cells in the orobranchial epithelium were counted for each embryo. Error bars indicate s.d. (g–i) Wild-type sibling at 7-dpf. Dotted line: section plane in h-i. Putative NECs are present in the orobranchial epithelium (arrowheads; dashed box in i, magnified without DAPI in top right corner). 5-HT, serotonin; Gf, gill filament; Hyp, hypothalamus; Nt, neural tube; Obc, orobranchial cavity. Scale-bars: 50 μm in b,d,h; 25 μm in c,e,i.DOI:
http://dx.doi.org/10.7554/eLife.21231.005Figure 2—figure supplement 1. Putative NECs in the skin of embryonic zebrafish are not neural crest-derived.(a–b’) Three-dimensional rendering of the eye of a 3-dpf Tg(crestin:creERT2);Tg(-3.5ubi:loxP-GFP-loxP-mCherry) embryo, immunostained in whole-mount for serotonin and mCherry. Serotonergic cells are scattered in the epidermis over the eye, as reported (Coccimiglio and Jonz, 2012), but none is mCherry-positive, i.e., neural crest-derived (arrowheads). For associated z-stack movies, see Videos 1 and 2. Scale-bars: 100 μm in a; 50 μm in b.DOI:
http://dx.doi.org/10.7554/eLife.21231.006 Image published in: Hockman D et al. (2017) © 2017, Hockman et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View |