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Figure 2


Fig. 2. Hand2 immunoreactivity is intranuclear in developing enteric neurons but cytoplasmic when they mature. (A) Polyclonal antibodies raised against a 15-amino acid N-terminal sequence of Hand2 react with transfected Hand2-expressing (Hela H2) but not non-transfected Hela [Hela wild type (WT)] cells. (B) At E12, Hand2 antibodies react with crest-derived cells in the gut and prevertebral sympathetic ganglia of wild-type (WT) but not Wnt1-Cre-H2{Delta} fetuses. (C) Fetal gut dissociated at E11.5 and cultured overnight; the nuclei of Hu+ and PGP9.5+ neurons are Hand2+ (H2; red). (D) Fetal gut dissociated at E14 and cultured similarly to those in C; nuclei and cytoplasm are both Hand2+ (red). DNA is blue. (E) A dissected laminar preparation of adult bowel containing a submucosal ganglion viewed as a whole mount. Hand2 immunoreactivity (red) is restricted to the cytoplasm of Hu+ (green) neurons. DNA (blue) stained with bisbenzimide. Scale bars: 50 µm in A,B; and 25 µm in C-E.