Fig. 3. MO1 disrupts axial development in Xenopus and exerts
dose-dependent effects on gene expression in the early gastrula. (A-C)
Xenopus embryos were injected at the one-cell stage with mMO1 (A; 40
ng) or MO1 (B,C; 40 ng) and allowed to develop to tadpole stage 33. MO1 causes
axial defects and a disruption of anterior development. (D-G) The phenotype
illustrated in B,C is presaged by disruption of expression of Xnot
(D,E) and Chordin (F,G). Xnot expression persists around the
blastopore of embryos injected with MO1. (E) Expression of Chordin in
embryos injected with MO1 (F) is more diffuse than that in embryos injected
with mMO1 (G). (H-K) Although axial morphogenesis is disrupted in embryos
injected with antisense morpholino oligonucleotide MO1, notochord and muscle
do nevertheless form in such embryos. (H,J) Embryos injected with mMO1 stained
with monoclonal antibody MZ15 (H) or 12/101 (J). (I,K) Embryos injected with
MO1 stained with MZ15 (I) or 12/101 (K). (L) Embryos were injected with the
indicated amounts of MO1 and allowed to develop to early gastrula stage 10.5,
when gene expression was assessed by real-time RT-PCR. Levels of gene
expression are normalised to those of ornithine decarboxylase.
Increasing concentrations of MO1 cause the downregulation first of dorsally
expressed genes such as Goosecoid and chordin and then the
downregulation of Xbra, which is expressed throughout the marginal
zone. Expression of Xwnt8, which occurs in lateral and ventral
tissue, is little affected.