Fig. 2: Posterior and anterior lineage analysis of pentadactyl and Grem1 tetradactyl forelimb buds. | Nature Communications

Fig. 2: Posterior and anterior lineage analysis of pentadactyl and Grem1 tetradactyl forelimb buds.

From: Single-cell profiling of penta- and tetradactyl mouse limb buds identifies mesenchymal progenitors controlling digit numbers and identities

Fig. 2: Posterior and anterior lineage analysis of pentadactyl and Grem1 tetradactyl forelimb buds.

a Tracing the posterior lineage by ShhCRE-mediated permanent activation of the ROSA26tdTomato reporter (magenta). Representative forelimb buds are shown. Left panel: wild-type forelimb bud at E11.5–E11.75 (n = 6/6). Arrowhead indicates the anterior boundary of the posterior lineage. Middle and right panels: wild-type forelimb buds at E12.5 (n = 6/6). Enlargements show optical sections to better reveal the posterior lineage between digits d2 and d4. b Tracing the posterior lineage in tetradactyl E1C5Δ/Δ forelimb buds. Left panel: E1C5Δ/Δ forelimb bud at E11.5–E11.75. Arrowhead indicates the anterior boundary of the posterior lineage, which is shifted in comparison to the wild type (n = 3/4). Middle and right panels: E1C5Δ/Δ forelimbs at E12.5. Enlargements show optical sections to better reveal the posterior lineage between digits d3* and d4. The lineage extends to the middle of d3* (n = 5/6). c Tracing the anterior lineage by tamoxifen-mediated induction of the Alx4-CreERT2 transgene around E10.5, which activates the Rosa26eGFP reporter (green). Representative forelimb buds are shown. Left panel: wild-type forelimb bud at E11.5–E11.75 (n = 6/6). Arrowhead indicates the posterior boundary of the anterior lineage. Middle and right panels: wild-type forelimb at E12.5 (n = 5/6). Enlargements show optical sections to better reveal the anterior lineage. d Anterior lineage analysis of tetradactyl E1C5Δ/Δ forelimb buds. Left panel: E1C5Δ/Δ forelimb bud at E11.5–E11.75 (n = 5/5). Arrowhead indicates the posterior boundary of the anterior lineage. Middle and right panels: E1C5Δ/Δ forelimb at E12.5–75 (n = 5/6). Enlargements show optical sections to better reveal the region between digit d3* and d4. Scale bars for (a–d): 1st panel from the left, 200 μm, 2nd and 4th panels, 500 μm, and 3rd and 5th panels (insets), 250 μm. e Virtual merge of the posterior and anterior lineages from two different limb buds at E12.5–75 (n = 2). Left and middle panels: single gray channels showing the anterior and posterior lineages after transformation. Right panels: the virtual merge of the anterior and posterior lineages. Scale bar: 300 µm. All limb buds are oriented with anterior to the top and posterior to the bottom, proximal to the left, and distal to the right.

Back to article page