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Fig. 7. NFPCMO and TAF1MO alter the organization of the neural folds. (A�F)
Embryos were co-injected with NFPCMO (A, D), TAF1MO (B, E), or CMO
(C, F), and nlacZ RNA, fixed at stage 17, processed for β-galactosidase activity,
then stained with Oregon green-phalloidin to visualize F-actin, and viewed
under fluorescence (A�C) or bright field (D�F). While F-actin was localized to
the apical neural folds in CMO-injected embryos as well as on the uninjected
side in NFPCMO- and TAF1MO-injected embryos (closed arrows in panels
A�C), F-actin staining was lost at the tips of the neural folds on the injected
side (open arrows in panels A, B). (G�L) Embryos injected with antisense
morpholinos and examined by immunofluorescence for β-catenin localization at
stage 17. In control embryos and on the uninjected side of NFPCMO- and
TAF1MO-injected embryos, the neural folds formed normally. In contrast, the
neural fold on the NFPCMO- or TAF1MO-injected side formed abnormally.
High power photomicrographs revealed a disorganized neural epithelial on the
injected side of NFPCMO- and TAF1MO-injected embryos (arrowheads in
panels J, K) as compared to the typical columnar epithelia observed on the
uninjected side or in control embryos (long arrows in panels J�L). In all cases,
β-catenin was still localized to the cell membrane. In all photos, the injected side
is on the right. Abbreviations: nf, neural fold, nc, notochord, nt, neural tube. |