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Fig. 2. Wnt3a protein re-specifies chondrogenic cells towards soft connective
tissues. (A,B) Micromass cultures from chick limb mesenchyme
cells treated with vehicle (A) or Wnt3a (B) for 4 days, cultured another 4
days in absence of Wnt3a. Wnt3a-treated cells failed to undergo chondrogenesis
(Alcian Blue; n>50). (C) Collagen 1, tenascin C, decorin,
Dermo1 and Bmp3 expression levels in high density cultures grown in continuous
presence of Wnt3a (blue), or during the first 3 days of culture, after which
the Wnt3a was replaced by vehicle (red). Time points 1 and 8 days were sampled
twice (mean±s.e.m.). (D) Small numbers of myotubules,
immunostained for myosin heavy chain (brown), accumulate in the periphery of
6-day-old mouse limb mesenchyme micromass cultures (n=6). (E)
Continuous treatment with Wnt3a (250 µg/ml) slightly expands the myotubule
number (n=6). (F) When Wnt3a is removed after 3 days of
culture, large numbers of myotubules spread over the tissue layer
(n=6). (G) Section through a chick wing 3 days after
implantation of a vehicle bead at embryonic stage 22, showing the bead
embedded in cartilage (n=14, Safranin O). (H) Wnt3a beads are
never embedded in cartilage (n=14) but surrounded by ectopic muscle
fibers, visualized by myosin heavy chain immunostaining (brown, n=7).
(I,J) Section through a control chick wing (I) and a wing with
implanted Wnt3a bead (J), immunostained for pro-collagen 1 (red) and myosin
heavy chain (green), nuclei stained blue (DAPI) (n=4). Both sections
are at a similar location and plane. Scale bars: 500 µm in A,B; 100 µm
in G-J. B, bead; car, cartilage.
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