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Fig. 9. Profilin1 is specifically required for blastopore closure.
(A) Schema of the injection approach to overexpress or deplete
Profilin1 in or lateral to the neural folds, adapted from Wallingford and
Harland (Wallingford and Harland,
2002). RNA (50 pg) encoding membrane-localized GFP injected into
the lateral blastomeres at the 16-cell stage targets expression lateral of the
neural folds whereas injection into the medial blastomeres targets expression
to the neural folds. (B) Targeted overexpression of Profilin1 RNA (2
ng) or depletion of Profilin1 (50 ng MO) in the neural folds does not inhibit
anterior neural fold closure but leads to open neural folds in the posterior,
reflecting a failure of blastopore closure. This phenotype can be rescued by
coexpression of 
N-Profilin1 (50 pg). Lateral overexpression or
depletion of Profilin1 has no effect on neural fold closure. (C) Time
lapse images of embryos injected with Profilin1 RNA (2 ng) or XProfilin1 MO
(50 ng) showing a delay and eventual failure of blastopore closure. The
XProfilin1 MO phenotype is rescued by coinjection of 50 pg N-Profilin1
RNA. (D) Depletion of XProfilin1 and XDaam1 synergistically inhibit
gastrulation. Quantitation of the phenotypes observed with separate or
co-injections of XProfilin1 MO and/or XDaam1 MO, Embryos with an open
blastopore and significantly reduced anterior posterior (AP) axis were scored
as severe embryos. Embryos with a small open blastopore or delayed blastopore
closure and a shortened AP axis or bent body axis were scored as intermediate
to mild. (E) A model for how Profilin1 functions in the non-canonical
Wnt signaling pathway (see Discussion).
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