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Fig. 3. Lrp4 is required for neuromuscular synaptogenesis. (A) Newborn
wild-type pups display motile activity and have begun feeding shortly after
birth (indicated by the presence of a milkspot). By contrast, Lrp4
mutant pups (B) are cyanotic, do not move, feed or breathe and retain
their in utero posture. (C,D) Longitudinal histological sections
through the left lung lobe of E18.5 wild-type (C) and mutant embryos (D). The
future alveolar spaces (al) are smaller and the interstitial mesenchyme is
thicker in Lrp4 mutant (D) embryos compared with wild type (C). E18.5
intercostal (E,F) and hind limb (G,H) muscles
stained with 
-bungarotoxin (red) to mark acetylcholine receptors and
co-visualized with HB9-GFP (E-F, green), which is expressed specifically in
motoneurons, or synaptophysin (G-H, green) which marks synaptic vesicles.
E18.5 wild-type (I-K) and mutant (L-N) diaphragm muscles stained
with -bungarotoxin (red) and co-visualized with HB9-GFP (green).
Lrp4 mutants show extensive branching of the phrenic nerve in
diaphragms and lack AChR clustering in the synaptic endplate (N). In all types
of muscle examined, Lrp4 mutants lack normal AChR clustering in the
synaptic endplate.
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