FIGURE 3

FIGURE 3

Axonal branching and overextension in fat2 mutants. (a–k) Expression of Vglut1 (a–c, g–i) and parvalbumin7 (Pvalb7, d–f, g–i) in 5‐dpf wild‐type (n = 5), fat2^{nub128/nub128} (n = 5), and fat2^vu381/vu381 mutant (n = 5) larvae. Neurod1 expression in 5‐dpf wild‐type (n = 5) and fat2^{nub128/nub128} mutant (n = 5) larvae (j, k). Signals were detected by immunostaining. Fluorescence signals are shown in grayscale (a–f, j, k) or as two‐color images (g–i; Pvalb7, magenta; Vglut1, green). Dorsal views with anterior to the left. Arrowheads indicate branched axon bundles, and arrows indicate axons extending beyond the crest cell region (overextension). Asterisks indicate crest cells. (l, m) Number of branched axon bundles per larva in fat2^{nub128/nub128} (l) and fat2^vu381/vu381 (m) mutants compared with wild‐type larvae. (n, o) Total length of misprojecting axons per larva, measured from branching points to their distal ends, in fat2^{nub128/nub128} (n) and fat2^vu381/vu381 (o) mutants compared with wild‐type larvae. (p, q) Number of Pvalb7⁺ Purkinje cells per larva in fat2^{nub128/nub128} (p) and fat2^vu381/vu381 (q) mutants compared with wild‐type larvae. (r) Number of Neurod1⁺ granule cells per larva in fat2^{nub128/nub128} mutants compared with wild‐type larvae. Welch's t‐test. **p < 0.01; ***p < 0.001; ns, not significant. Scale bar: 100 μm in (a) (applies to a–i), and (j) (applies to j, k). Regions of the cerebellum, including CCe, EG, and LCa, are indicated in the images. Images are representative of the samples examined.