XB-IMG-172691
Xenbase Image ID: 172691
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Fig. 6. PFKFB4 morphant NC progenitors fail to undergo EMT and
migration. (A,B) When transplanted into a wild-type host embryo, GFPlabeled
wild-type NC efficiently migrates and populates the host branchial
arches, regardless of the size of the grafted tissue, i.e. large graft (A) or small
graft (B). Scale bar: 500 μm. (C,D) In contrast, PFKFB4 morphant NCexhibited
defective healing, resulting in small-size grafts, which yielded few, if any,
migratory NCC into the host craniofacial area. (C) Small graft without migratory
cells; (D) small graft with few cells reaching the branchial arches. In D, gfp was
detected by WISH to enhance individual cell visualization. (E) The effect of
PFKFB4MO on N-cadherin expression (RT-qPCR on dissected explants),
either prior to EMT (NB, st.14), or in premigratory NC (st.17). Error bars
represent s.e.m. *P≤0.05. (F-J) Wild-type (F,G; gfp only, no MO) or morphant
(G,I; MO 10 ng) cells were lineage traced in vivo, on embryos injected in the
prospective neural fold unilaterally. At tailbud stage, control cells (G) efficiently
populated branchial arches and were also found in brain and eye (J). In
contrast, morphant cells failed to populate branchial arches (I) in a dosedependent
manner (J), but were normally distributed in the brain and eye (J).
(K-R) NB tissue was dissected prior to EMT (st.14) and plated onto fibronectin.
Cell behavior was followed by time-lapse videomicroscopy.Wild-type NC (K-N)
adheres efficiently, and undergoes EMT (L), cell scattering and migration (M,
N). Morphant NC (O-R) presented poor adherence, delayed (P) and inefficient
EMT (Q), and few emigrating cells (R). Time after plating is indicated (h:min).
Scale bar: 160 μm. Phenotype scores are indicated in Table S10. 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. Larger Image Printer Friendly View |