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Fig. 4. lbx1 is required for hypaxial muscle formation. RT-PCR was
performed on tadpoles injected with lbx1-splice MO into one out of
two cells or both cells at the two-cell stage. (A) A specific loss of
the properly spliced lbx1 transcript is seen. Embryos were injected
into one cell at the two-cell stage with a control MO (B-E), the
lbx1-splice blocking MO (F-I) or the
lbx1-translation blocking MO (J-M) along with
ß-galactosidase mRNA as a lineage tracer (red). Tadpoles were stained
with the 12/101 antibody (brown). The uninjected (B,D,F,H,J,L) and injected
(C,E,G,I,K,M) sides of each tadpole are shown, where the uninjected sides are
pictured with anterior towards the right. The control MO-injected tadpoles do
not show a difference in the presence of rectus abdominus muscles on the
injected side compared with the uninjected side at stage 37 (compare B with C)
or stage 40 (D compared to E). Tadpoles injected with the lbx1-splice
MO exhibit a loss of rectus abdominus and rectus cervicus muscles on the
injected side at stage 37 (compare F with G). At stage 40, these muscles
appear but are greatly reduced in the lbx1-splice MO-injected
tadpoles (compare H with I). Similar results were observed for the lbx1-trans
MO-injected tadpoles (J-M). (N,O) Stage 40 control injected (N)
and lbx1-splice injected (O) embryos are shown from a ventral view,
with the MO injected side downwards (embryos were injected into one cell at
the two-cell stage). The geniohyoideus muscle is lost on the
lbx1-splice injected side of the tadpole (O, arrow), whereas both are
present in the control injected MO (N, arrow). Myoblast migration defects are
observed in lbx1-splice injected tadpoles. (P-S)
Tadpoles in P and R are stained for pax3 mRNA (purple) and then
merged with the 12/101 antibody (green, Q,S). A control tadpole is pictured in
P and Q, which shows the movement of pax3-positive myoblasts away
from the differentiated epaxial myotome. In lbx1-splice-injected
tadpoles (R,S), a large number of pax3-positive myoblasts remain
adjacent to the differentiated myotome.
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