ZFIN ID: ZDB-FIG-080516-45
Minchin et al., 2008 - Sequential actions of Pax3 and Pax7 drive xanthophore development in zebrafish neural crest. Developmental Biology   317(2):508-522 Full text @ Dev. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Knockdown Reagents:
Anatomical Terms:
Stage: Prim-5
PHENOTYPE:
Fish:
Knockdown Reagents:
Observed In:
Stage: Prim-5

Fig. 3 Pax3 morphants have a defect at early stages of xanthophore development. Control (A, C, E, G, I, K, M, O, Q, S, U, W) and pax3 MO-injected (B, D, F, H, J, L, N, P, R, T, V, X) 1-day-old zebrafish embryos analysed by in situ mRNA hybridisation for xdh (A–D), csf1r (E–H), mitfa (I, J), gch (K–N) or pax7 (Q–T) or by immunodetection for Pax3/7 with DP312 (O, P) or anti-Pax7 (U–Y). Lateral views anterior to left. Insets show dorsal views of the head region. (A–D) Pax3 MO essentially ablates xdh mRNA in trunk (arrows), but reduced expression persists in the head (arrowheads). (E–H) Csf1r mRNA is undetectable in pax3 morphant trunk. Although residual expression is present in pax3 morphant head, it appears differently localised compared to xdh (white arrowheads). (I, J) In control embryos, expression of the chromatophore marker mitfa shows the start of NC migration in anterior trunk. In pax3 morphants, mitfa expression is severely reduced, but less so than xdh and csf1r. (K–N) Gch mRNA is strongly depleted in trunk of pax3 morphants (arrows), but is still detectable in head (arrowhead). (O, P) At 25 hpf, DP312 detects cells in the dermomyotome weakly (arrowheads) and pre-migratory NC cells intensely (arrows). In pax3 morphants, the strongly-labelled NC cells are dramatically reduced, whereas dermomyotome signal remains. Note the co-localisation of Pax3/7 and Hoechst 33258 DNA stain in transverse cryosections of one side of the trunk at right. (Q–T) At 25 hpf, pax7 mRNA is lost from trunk NC cells, but retained in dermomyotome and neural tube. (U–X) Immunodetection of Pax7 at 27 hpf demonstrates that intensely labelled cells migrate in an anterior to posterior wave on the lateral surface of the somite, resembling NC. Pax7+ cells are severely reduced or absent in the trunk of pax3 morphants, and mildly affected in head regions (insets). (Y) Counts of strongly Pax7+ NC cells quantify the difference in severity between head and trunk regions.

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
aox5 WT control Prim-5 cranial neural crest cell ISH
Prim-5 trunk neural crest cell ISH
WT + MO3-pax3a standard conditions Prim-5 cranial neural crest cell ISH
Prim-5 Not Detected trunk neural crest cell ISH
WT + MO4-pax3a standard conditions Prim-5 cranial neural crest cell ISH
Prim-5 Not Detected trunk neural crest cell ISH
csf1ra WT control Prim-5 cranial neural crest cell ISH
Prim-5 trunk neural crest ISH
WT + MO3-pax3a standard conditions Prim-5 cranial neural crest cell ISH
Prim-5 Not Detected trunk neural crest cell ISH
WT + MO4-pax3a standard conditions Prim-5 cranial neural crest cell ISH
Prim-5 Not Detected trunk neural crest cell ISH
gch2 WT control Prim-5 cranial neural crest cell ISH
Prim-5 trunk neural crest cell ISH
WT + MO3-pax3a standard conditions Prim-5 cranial neural crest cell ISH
Prim-5 Not Detected trunk neural crest cell ISH
WT + MO4-pax3a standard conditions Prim-5 cranial neural crest cell ISH
Prim-5 Not Detected trunk neural crest cell ISH
mitfa WT control Prim-5 trunk neural crest cell ISH
WT + MO3-pax3a standard conditions Prim-5 trunk neural crest cell ISH
WT + MO4-pax3a standard conditions Prim-5 trunk neural crest ISH
pax7a WT control Prim-5 dermomyotome ISH
Prim-5 neural tube ISH
Prim-5 trunk neural crest cell ISH
WT + MO3-pax3a standard conditions Prim-5 dermomyotome ISH
Prim-5 neural tube ISH
Prim-5 Not Detected trunk neural crest cell ISH
WT + MO4-pax3a standard conditions Prim-5 dermomyotome ISH
Prim-5 neural tube ISH
Prim-5 Not Detected trunk neural crest cell ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
WT + MO3-pax3a standard conditions Prim-5 head xanthophore decreased amount, abnormal
Prim-5 trunk xanthophore absent, abnormal
Prim-5 xanthophore differentiation disrupted, abnormal
WT + MO4-pax3a standard conditions Prim-5 head xanthophore decreased amount, abnormal
Prim-5 trunk xanthophore absent, abnormal
Prim-5 xanthophore differentiation disrupted, abnormal
Acknowledgments:
ZFIN wishes to thank the journal Developmental Biology for permission to reproduce figures from this article. Please note that this material may be protected by copyright.

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.