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Experiment details for sox10

PFKFB4 control of AKT signaling is essential for premigratory and migratory neural crest formation.

PFKFB4 control of AKT signaling is essential for premigratory and migratory neural crest formation.

Gene Clone Species Stages Anatomy
sox10.L laevis NF stage 18 neural crest , neural plate border
sox10.L laevis NF stage 24 mandibular crest , hyoid crest , otic placode , cranial neural crest , migratory neural crest cell

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  Fig. 3. PFKFB4 low-level depletion delays NC early specification, causes retention of NB character, and impairs NC late specification and migration. (A) At st.14, snail2 expression was severely reduced, or abolished, on the injected side. (B) At st.18, sibling embryos had recovered snail2 expression. (C,D) Whereas sox10 expression was mainly unaffected, twist1 was severely impaired. (E-H) In contrast, expression of the immature NC marker hes4 was expanded, as were some NB markers, either strongly ( pax3) or moderately (zic1). Other NB markers were unperturbed (msx1). (I-K) Neural plate (sox2), non-neural ectoderm (ep. ker.) and paraxial mesoderm (myod) seemed to be unaffected. A-K: dorsal views. (L) Percentage of embryos with each phenotype, i.e. diminished, increased or normal expression. Snail2 score at st.14 is indicated as the first bar, then several gene scores at st.18 are indicated in the following ten bars. (M-R) St.24 tailbud embryos exhibited a severe NC migration defect (M-Q). Sox2 expression appeared grossly unaffected, despite marginal reduction of optic vesicle size (R). The injected side (inj) is compared with the control side (co) in side views (M,N,P,Q, anterior to the right) or frontal views (O,R; red arrow on injected side). (S) Co-injection of pfkfb4 mRNA with PFKFBMO rescued both sox10/twist1 alterations of expression and NC migration defects in a significant proportion of the embryos, compared with PFKFB4MO injections alone. sox10 and twist1 expression were restored or increased a majority of the embryos. Scale bar: 500 μm. Phenotype scores are shown in Table S8.

Gene Clone Species Stages Anatomy
sox10.L laevis NF stage 24 mandibular crest , hyoid crest , otic placode , cranial neural crest , migratory neural crest cell

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  Fig. 7. Glycolysis and PI3K-AKT signaling impact NC migration similarly to PFKFB4 low-level depletion, and activating AKT signaling rescues PFKFB4 downregulation. (A-H) When glycolysis (2DG) or PI3K-AKT pathway (LY294002) were blocked during EMT and migration (st.18-24) both treatments severely affected NC migration at st.24 (as revealed by twist1 and sox10 expression). (I-L) At st.45, tadpoles treated during NC migration then grown in control medium, exhibited general head morphology defects, including eye defects and smaller branchial cartilages. (I,K) Sibling controls; (J) 2DG; (L) LY294002. A-H show side views; I-L dorsal views. (M-P) At tadpole st.45, morphant sides were severely affected (M,P), whereas activation of Akt signaling (N,P) did not affect overall craniofacial morphogenesis. Tadpoles coinjected with PFKFB4MO and caAkt (O,P) were largely rescued, with 66% of embryos with injected side symmetrical to contralateral side. (M) Red bar indicates eye distance from the midline on the morphant side; blue bar indicates control distance. Both bars are aligned below for comparison. (N,O) On both sides, the same blue bar measures eye distance from the midline. Arrowheads indicate the injected side. Scale bar: 500 μm. Phenotype scores are shown in Tables S11, S12.

Gene Clone Species Stages Anatomy
sox10.L laevis NF stage 26 mandibular crest , hyoid crest , cranial neural crest , migratory neural crest cell

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  Fig. S6. Glycolysis and PI3K-Akt signaling control neural crest migration. Glycolysis blockade (AJ) and PI3K-Akt blockade (K-T) during EMT and neural crest migration (stage 18 to stage 24) affects neural crest migration as shown by the expression of Sox9, Sox10 and Twist. Embryos rinsed at stage 24 and grown until stage 45 display craniofacial and eye development defects, albeit the presence of all individual cartilage elements. (A-C,F-H,K-M,P-R, U-W) Side views, anterior is to the left. (D,I,N,S,X) dorsal views st 45. (E,J,O,T,Y) Ventral views of alcian blue stained visceral cartilages, st 45. Scale bar= 500μm.