Larger Image 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. Image published in: Figueiredo AL et al. (2017) Copyright © 2017. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License.
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