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Fig. 1. PFKFB4 promotes NB and NC formation.
(A) NC is induced during neural plate stage (st.12.5-14).
Expression of NB specifiers (red) is robustly established
lateral to the neural plate, expression of early NC
specifiers (snail2, foxd3) is faintly initiated (light pink).
(B,C) During neural fold elevation (st.14-17, B), future
NC cells progressively acquire their definitive
specification and specific cellular properties (e.g.
survival, expression of late NC specifiers, cadherin
switch) enabling them to undergo EMT and migration
(st.18-19, C). (D-F,H,I) pfkfb4 is enriched in the NB/NC in
neurulas and tadpoles. (J) pfkfb3 is expressed in the
neural tube. Sense probes are shown inG( pfkfb4) and K
( pfkfb3). Neurula stages: late gastrula/early neural plate
stage (st.12, D); end of neural plate stage (st.14, E);
neural fold stage (st.16, F); neural tube closure/end of
neurulation (st.19, H); tailbud stage (st.22, I).
E-H,J,K: dorsal views, anterior to the right. D,I: side
views. Scale bar: 1 mm. (L-N) Cross-sections through
the mid-neurula anterior neural plate (st.14) show snail2
and pfkfb4 expression in NB/NC. (L) The notochord,
neural plate and paraxial mesoderm are outlined on
Hematoxylin and Eosin-stained sections. (M,N)
Vibratome sections. Arrow indicates the midline.
Arrowheads indicate the st.14 NB (red bar). Scale bars:
200 μm. (O-S) pfkfb4 gain-of-function expanded the
st.14 NB (O,P) and premigratory NC (st.14, Q; st.18, R,
S). Migrating NC streams were expanded (st. 22-24, T,
Z). In contrast, pan- or regional neural plate (sox2, otx2,
hoxb9), ectoderm (ep. ker.) or paraxial mesoderm
(myod) markers were unaffected (U-Y). O-S,U-X: dorsal
views; T,Z: side views; Y: dorsal-posterior view. Scale
bar: 500 μm. co, control side; inj, injected side.
Phenotype scores are shown in Table S6. |
