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Fig. 1. The numbers of NCSCs, neurons and glia are reduced in the gut after
Rbpsuh deletion. (A,B) Transverse sections of the
guts from E13.5 control (A) or Wnt1-Cre+
Rbpsuhfl/fl (B) mouse embryos were stained for neurons (TuJ1)
and glia (BFABP). Scalebar: 10 µm. (C) The numbers of
p75+ neural crest cells that initially colonized the gut did not
differ between E10.5 control and Wnt1-Cre+
Rbpsuhfl/fl embryos (the number of p75+ cells per
section; three sections per gut region per mouse, three mice per genotype).
(D-F) At representative levels of the developing gut (foregut, midgut
and hindgut), the numbers of neurons (TuJ1+) and glia
(BFABP+) per section were similar to wild type in
Wnt1-Cre+ Rbpsuhfl/fl embryos at E10.5 (D) but
significantly (*, P<0.05) lower at E14.5 (E) and E18.5
(F). (G) There was no difference in the rate of proliferation of
p75+ cells at E14.5 between control and mutant embryos (BrdU was
administered for 30 minutes; three to seven sections per gut region per mouse,
three to four mice per genotype). Additionally there was no significant
difference in the rate of cell death between control and mutant mice at any
stage of development (E10.5, E14.5 and E18.5) or at any level of the gut
(foregut, midgut and hindgut) based on staining for activated caspase 3 (data
not shown). (H) However, a significantly (*,
P<0.05) lower percentage of cells from E13.5
Wnt1-Cre+ Rbpsuhfl/fl guts gave rise to
multilineage NCSC colonies in clonal-density cultures in both standard medium
and medium supplemented with neuregulin (+Nrg) (three to seven independent
experiments). All error bars indicate s.d.
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