FIGURE

Fig. 7

ID

ZDB-FIG-080516-49

Publication

Minchin et al., 2008 - Sequential actions of Pax3 and Pax7 drive xanthophore development in zebrafish neural crest

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Fig. 7

Delayed melanophore origin then increase correlate with xanthophore loss in pax3 morphants. Embryos analysed by in situ mRNA hybridisation for dct (A–F), live (G–J) or fixed (K–N) shown with anterior to left. Wholemounts are shown in lateral (A, B, E–J; dorsal to top) or dorsal (K–N) view. Whole embryo flatmounts (C, D) show dorsal and lateral views in head and trunk regions, respectively. (A, B) At 25 hpf, dct mRNA is detected in pre-migratory (arrows) and migrating melanoblasts (arrowheads) in the head and trunk, and also in the eye pigment epithelium. In pax3 morphants, dct⁺ melanoblasts are reduced, but still able to migrate (B, arrowhead). (C, D) At 35 hpf, substantially more dct⁺ melanoblasts are situated dorsally in pax3 MO2-injected embryos (arrows), but migrated dct⁺ melanoblasts are reduced (arrowheads). (E–H) At 48 hpf, dct mRNA is present in dorsal, lateral and ventral stripes, and in single cells on yolk sac, matching the melanophore pattern (E, G red arrows). In pax3 morphants, melanised and dct⁺ cells are more abundant between stripes on the medial and lateral pathways (F, H arrowheads). (I, J) At 5 dpf in control embryos, four larval melanophore stripes are clearly defined (I, red arrows) and the trunk has a yellow/green cast due to xanthophore pigmentation (I, asterisk). In pax3 morphants, the trunk is devoid of xanthophore pigmentation (J, asterisk), but the melanophore pattern is within normal variation (J, red arrows). (K-N) Treatment of 6 dpf embryos with adrenalin causes melanophore contraction that permits cell counting. In trunk (K, L) but not head (M, N anterior to red bar), pax3 MO-injection increases melanophore number in the dorsal stripe. (O) Quantification of melanophore number in pax3 MO, pax3 MO2 and pax7 MO-injected embryos at 6 dpf. pax7 morphants were categorised into normal yellow pigmentation (+ YP) or reduced yellow pigmentation (- YP) groups. ***p < 0.0001. (P) Correlation of melanophore increase in pax3 morphants with loss of xanthophores. Pax3 MO2-injected 8 dpf larvae were categorised by severity of methylene blue-stained xanthophore phenotype, and melanophores counted. DS = dorsal stripe, LS = lateral stripe, H = head melanophores.

Expression Data

Gene:	dct
Fish:	WT WT + MO3-pax3a WT + MO4-pax3a
Knockdown Reagents:	MO3-pax3a MO4-pax3a
Anatomical Terms:	extension head melanoblast melanocyte melanophore stripe presumptive retinal pigmented epithelium retinal pigmented epithelium trunk
Stage Range:	Prim-5 to Long-pec

Expression Detail

Antibody Labeling

Phenotype Data

Fish:	WT + MO3-pax3a WT + MO4-pax3a
Knockdown Reagents:	MO3-pax3a MO4-pax3a
Observed In:	head melanocyte melanoblast melanocyte differentiation melanophore stripe melanophore stripe melanocyte trunk melanocyte trunk xanthophore
Stage Range:	Prim-5 to Day 6

Phenotype Detail

Acknowledgments

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Reprinted from Developmental Biology, 317(2), Minchin, J.E., and Hughes, S.M., Sequential actions of Pax3 and Pax7 drive xanthophore development in zebrafish neural crest, 508-522, Copyright (2008) with permission from Elsevier. Full text @ Dev. Biol.