ZFIN ID: ZDB-FIG-120216-32
Balczerski et al., 2012 - Analysis of Sphingosine-1-phosphate signaling mutants reveals endodermal requirements for the growth but not dorsoventral patterning of jaw skeletal precursors. Developmental Biology   361(2):230-241 Full text @ Dev. Biol.
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Condition:
Anatomical Terms:
Stage Range: Prim-15 to Prim-25
PHENOTYPE:
Fish:
Condition:
Observed In:
Stage Range: Prim-15 to Prim-25

Fig. 6

Shha misexpression partially rescues the jaw defects of sox32 mutants. (A–D) Ventral views of 6 dpf head skeletons. Compared to hsp70I:Gal4 controls subjected to the same heat-shock treatment (A), hs-Shha larvae developed ectopic Tr-like cartilages (arrowheads) (B, n = 10/10). Whereas facial skeleton never formed in non-hs-Shha sox32 siblings (C, n = 0/10), jaw and hyoid cartilage was partially restored and ectopic cartilage was still present in sox32; hs-Shha larvae (D, n = 4/4). Flat-mount preparations (insets in A and D) show that Meckel′s (M), palatoquadrate (Pq), and hyosymplectic (Hs) cartilages, but not the hyoid-arch-derived ceratohyal (Ch) cartilage, were dysmorphic yet present in sox32; hs-Shha larvae. (E–H) In situ hybridizations show fgf8a oral ectoderm expression (arrows) at 36 hpf. Compared to hsp70I:Gal4 controls (E), fgf8 expression was shifted dorsal-anteriorly in hs-Shha embryos (F, n = 8/9) and partially (n = 3/34) or severely reduced (n = 31/34) in sox32 mutants (G). Shha misexpression resulted in increased and dorsal-laterally shifted fgf8a expression in sox32; hs-Shha embryos (H, n = 11/12). (I–L) Confocal projections show the pharyngeal arches (numbered) of control fli1a:GFP (I, n = 3), hs-Shha; fli1a:GFP (J, n = 4), sox32; fli1a:GFP (K, n = 10), and sox32; hs-Shha; fli1a:GFP (L, n = 7) embryos at 30 hpf. In sox32 and sox32; hs-Shha embryos, the lack of endoderm resulted in arches 3–7 remaining as a single mass. White outlines show the maxillary (max), mandibular (mand), and hyoid domains for wild type. (M–T) Anti-pH3 staining marks proliferating cells (red, M–P), Lysotracker marks regions of cell death (red, Q–T), and fli1a:GFP labels CNCCs (green) of the mandibular (1) and hyoid (2) arches in control (M and Q), hs-Shha (N and R), sox32 (O and S), and sox32; hs-Shha (P and T) embryos at 36 hpf. n = 10 for each genotype. Scale bars = 50 μm. (U) Quantification of arch domain area and numbers of pH3- and Lysotracker-positive cells per arch area in control hsp70I:Gal4 (con), hs-Shha, sox32, and sox32; hs-Shha embryos. Brackets indicate comparisons showing statistical significance with a Tukey–Kramer HSD test (α = 0.05).

Gene Expression Details
Gene Antibody Fish Conditions Stage Anatomy Assay
EGFP el137Tg; kca4Tg/+; y1Tg heat shock Prim-15 to Prim-25 pharyngeal arch IFL
sox32ta56/ta56; el137Tg; kca4Tg/+; y1Tg heat shock Prim-15 to Prim-25 pharyngeal arch IFL
sox32ta56/ta56; y1Tg standard conditions Prim-15 to Prim-25 pharyngeal arch IFL
y1Tg standard conditions Prim-15 to Prim-25 pharyngeal arch IFL
fgf8a el137Tg; kca4Tg/+ heat shock Prim-25 oral region ectoderm ISH
kca4Tg/+ heat shock Prim-25 oral region ectoderm ISH
sox32ta56/ta56; el137Tg; kca4Tg/+ heat shock Prim-25 oral region ectoderm ISH
sox32ta56/ta56; kca4Tg/+ heat shock Prim-25 oral region ectoderm ISH
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
el137Tg; kca4Tg/+ heat shock Prim-25 cranial cartilage mislocalised, abnormal
Prim-25 cranium decreased size, abnormal
Prim-25 embryo development process quality, normal
Prim-25 oral ectoderm mislocalised anteriorly, abnormal
Prim-25 oral ectoderm mislocalised dorsally, abnormal
Prim-25 pattern specification process process quality, normal
el137Tg; kca4Tg/+; y1Tg heat shock Prim-15 - Prim-25 chondrocyte proliferation occurrence, normal
Prim-15 - Prim-25 maxilla increased size, abnormal
sox32ta56/ta56; el137Tg; kca4Tg/+ heat shock Prim-25 dorsal hyoid arch decreased size, abnormal
Prim-25 dorsal hyoid arch shape, normal
Prim-25 mandibular arch skeleton apoptotic, abnormal
Prim-25 Meckel's cartilage decreased size, abnormal
Prim-25 Meckel's cartilage shape, normal
Prim-25 oral ectoderm mislocalised dorsally, abnormal
Prim-25 oral ectoderm mislocalised laterally, abnormal
Prim-25 palatoquadrate cartilage decreased size, abnormal
Prim-25 palatoquadrate cartilage shape, normal
Prim-25 pharyngeal arch 2 skeleton structure, normal
Prim-25 pharyngeal arch 3-7 decreased size, abnormal
Prim-25 pharyngeal arch 3-7 fused with pharyngeal arch 3-7, abnormal
Prim-25 pharyngeal arch cartilage associated with pharyngeal arch cartilage, normal
Prim-25 ventral hyoid arch decreased size, abnormal
sox32ta56/ta56; el137Tg; kca4Tg/+; y1Tg heat shock Prim-15 - Prim-25 chondrocyte proliferation increased occurrence, abnormal
Prim-15 - Prim-25 dorsal hyoid arch size, normal
Prim-15 - Prim-25 maxilla decreased size, abnormal
Prim-15 - Prim-25 pharyngeal arch 3-7 decreased size, abnormal
Prim-15 - Prim-25 pharyngeal arch 3-7 fused with pharyngeal arch 3-7, abnormal
Prim-15 - Prim-25 ventral hyoid arch size, normal
Prim-15 - Prim-25 ventral mandibular arch size, normal
sox32ta56/ta56; kca4Tg/+ heat shock Prim-25 dorsal hyoid arch decreased size, abnormal
Prim-25 dorsal hyoid arch shape, abnormal
Prim-25 Meckel's cartilage decreased size, abnormal
Prim-25 Meckel's cartilage shape, abnormal
Prim-25 oral ectoderm hypoplastic, abnormal
Prim-25 palatoquadrate cartilage decreased size, abnormal
Prim-25 palatoquadrate cartilage shape, abnormal
Prim-25 pharyngeal arch 3-7 decreased size, abnormal
Prim-25 pharyngeal arch 3-7 fused with pharyngeal arch 3-7, abnormal
Prim-25 pharyngeal arch cartilage associated with pharyngeal arch cartilage, abnormal
Prim-25 ventral hyoid arch decreased size, abnormal
sox32ta56/ta56; y1Tg standard conditions Prim-15 - Prim-25 apoptotic process increased occurrence, abnormal
Prim-15 - Prim-25 dorsal hyoid arch decreased size, abnormal
Prim-15 - Prim-25 mandibular arch skeleton apoptotic, abnormal
Prim-15 - Prim-25 maxilla decreased size, abnormal
Prim-15 - Prim-25 pharyngeal arch 2 skeleton apoptotic, abnormal
Prim-15 - Prim-25 pharyngeal arch 3-7 decreased size, abnormal
Prim-15 - Prim-25 pharyngeal arch 3-7 fused with pharyngeal arch 3-7, abnormal
Prim-15 - Prim-25 ventral hyoid arch decreased size, abnormal
Prim-15 - Prim-25 ventral mandibular arch decreased size, 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, 361(2), Balczerski, B., Matsutani, M., Castillo, P., Osborne, N., Stainier, D.Y., and Crump, J.G., Analysis of Sphingosine-1-phosphate signaling mutants reveals endodermal requirements for the growth but not dorsoventral patterning of jaw skeletal precursors, 230-241, Copyright (2012) with permission from Elsevier. Full text @ Dev. Biol.