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Figure 2. Characterization of the early, intermediate, and late phases of tail regeneration. (A) Tadpole tails imaged using bright field microscopy at early (0 h, 6 h); intermediate (24 h) and late phases (48 h and 72 h) of tail regeneration. Black open arrowhead shows typical dorsal constriction at 6 h post amputation; blue arrowhead shows pre-regenerative tissue; red arrowhead shows nascent fin epidermis at the distal tip; black arrowhead = spinal cord; green arrowhead = notochord, orange arrowhead = melanophrore. (B) Tadpole tails stained with Sudan Black B (inflammatory cells). (C) Tadpole tails stained by whole-mount in situ hybridization for mmp7 (inflammatory cells). (D) Tadpole tails stained by immunohistochemistry for mitotic cells (pH3, shown in black). The tail in each panel is outlined in red and the plane of amputation is shown in green. (E) Tadpole tails stained to reveal the neuronal tissue by immunohistochemistry (acetylated tubulin, shown in black). (F) Tadpole tails stained by immunohistochemistry for vascular tissue (GFP antibody in mTie-2::eGFP transgenic line, shown in black). The open purple arrowhead shows a typical eGFP positive "clot" at the injured dorsal aorta. Green arrowheads show distally projecting blood vessels. (G) Tadpole tails stained by immunohistochemistry for muscle tissue (12/101). The open orange arrowhead shows the presence of skeletal muscle in the regenerated portion of the tail. Red scale bar for each row is 250 μm. Note that the images in panels D-F are black and white reversals (i.e. shown as negative images). |
