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Fig. 5. Ventrally projecting motor axons displayed abnormal trajectories in
1.6MO morphants even though CaP axons project normally. (A-D) Motor
axon morphology was assayed using SV-2 immunoreactivity (green) in conjunction
with a marker of the muscle postsynaptic receptor, 
-bungarotoxin (red).
At 48 hpf (A,B), little difference was noted in znp-1 immunoreactivity of
motor axons of 1.6MO morphants (B) compared with control-injected embryos (A).
However, by 72 hpf, there were marked differences (C,D). Motor axons of 1.6MO
morphants (D, asterisks) branched more than those of control-injected embryos.
Moreover, in morphants (D), there was a reduction in the normal alignment
between motor axons and postsynaptic receptors, assessed by the near absence
of green and abundance of yellow in C versus the reduction in yellow and
increase in green in panel D. (E-L) Injection of 1.6MO did not affect
axon outgrowth from CaP when synapses first form. (E-H) At 21-22 hpf, znp-1
immunoreactivity revealed no differences between ventrally projecting CaP
axons in either control (E,F) or 1.6MO-injected (G,H) embryos. Embryos were
squash-mounted and, consequently, motor nerves on both sides of an embryo were
often present in a single confocal section (e.g. F,H). (I,J) CaPs were
directly labeled by dye injection (green) at 20 hpf and embryos were fixed at
21-22 hpf and stained for SV-2 immunoreactivity. Injection of either ConMO (I)
or 1.6MO (J) did not affect CaP axon morphology. (K,L) In addition to axonal
morphology (SV-2, green), postsynaptic specializations, as detected by
-bungarotoxin labeling (red) were not altered by injection of either
Con (K) or 1.6 (L) MO. Scale bars: in A, 20 µm for A,B; 30 µm for C; in
E, 50 µm for E-H; in J, 10 µm for I,J; in L, 20 µm for K,L.
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