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Fig. 4. Normal SOX10 activity and EDN3 signalling are required for formation of
NLBs and EPCs. Guts from E11.5 wild-type (A),
Sox10+/LacZ (B) or Ret51/51
(C) embryos were dissociated and cultured under conditions that promote
the formation of NLBs. After fixation, cultures were immunostained for TuJ1
(red) and GFAP (green), and counterstained with DAPI. NLBs form readily in gut
cultures from wild-type and Ret51/51 embryos but fail to
form in cultures from embryos hemizygous for Sox10. In wild-type gut
cultures maintained in standard medium (CM), NLBs form characteristic colonies
composed of undifferentiated cells, neurons and glia
(Bondurand et al., 2003). Such
a colony immunostained for TuJ1 (red) and GFAP (green) is shown (D).
These characteristic colonies do not appear in the presence of BQ788
(E). In the absence of CEE, embryo gut cultures failed to form NLBs
(F). However, addition of EDN3 was sufficient to rescue the formation
of NLBs (G) and EPCs capable of generating multilineage colonies. Part
of such an EPC colony is shown in the inset of G.
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