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Fig. 2. Effects of treatment with Shh and BMPs on the proliferation of granule cell precursors. Cerebellar granule cells were cultured in medium without additional growth factors (control), or with Shh (3 µg/ml), Bmp2 (100 ng/ml), Bmp4 (100 ng/ml) or Bmp7 (100 ng/ml). (A) After 60 hours in culture, cells were pulsed-labelled with [³H]thymidine for 12 hours, harvested and analysed for incorporation of radioactivity. BMPs had no effect on proliferation, whereas Shh induces a high rate of [³H]thymidine incorporation. (B-E) After 72 hour in culture, cells were immunostained for the neural-specific marker ßIII Tubulin (red) and the astroglial marker Gfap (green) in control cultures (B) and cultures treated with Shh (C), with Bmp2 (D) or with Bmp7 (E). Cultures treated with BMPs were not phenotypically different from control cultures. Shh-treated cultures showed the presence of big clumps of precursor cells that showed no immunoreactivity to either neural or glial marker. All cultures are counterstained with the nuclear marker DAPI (blue).

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