The temca mutation causes innervation defects in regenerating fins. (A) Schematic of method for assessing axon density and total axon length in 2 dpa fin regenerates. (B) Representative images of acetylated α-tubulin (ac-Tub) staining in 2 dpa control (heterozygotes) and temca fin regenerates at 33 °C. (C and D) Quantification of axon density (C) and total axon length (D) in a 150 × 200 µm2 area of control and temca 2-dpa fin regenerates at 33 °C. (E) Schematic of method for measuring axon bundle thickness and ratio in 2 dpa fin rays. (F–H) Representative images of ac-Tub staining (F) and quantification of axon bundle thickness (G) and ratio (H) in control and temca 2 dpa fin regenerates at 33 °C. (I) Schematic depicting angle of nerve bundle regrowth in 2 dpa fin regenerates. (J and K) Representative images of ac-Tub staining (J) and rose plots depicting orientation of regenerated nerve bundles (K) in control and temca 2 dpa fin regenerates at 33 °C (n = 8 and 12 for control and temca, respectively). (L) Representative whole-mount images of control and temca 7 dpa fins at 33 °C. (M) Representative images of ac-Tub staining in control and temca 7 dpa fin regenerates at 33 °C. White arrows indicate misshaped fin rays having misguided nerve bundles. White arrowheads indicate fin rays with complete blockage of regeneration. Insets display an enlarged version of the area in the white boxes. White dotted lines mark the distal tip of epidermis. Scale bars, 50 µm in B and J; 1.5 mm and 500 µm in L Top and Bottom, respectively; 250 µm in M. Data are presented as mean ± SD. Student’s unpaired two-tailed t test.
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