Fig. 4

Axon guidance defects in DINE mutant abducens nerves. Whole-mount fluorescent immunostaining with anti-neurofilament (red) and anti-GFP (green) antibodies of E11.5 wild-type (a), homozygous G607S mutant (b), and homozygous C760R mutant (c) embryos. White arrowheads represent the pathway of wild-type abducens nerves. d–f High-magnification views of E11.5 abducens nerves. The axon terminal (*) of mutant abducens nerves (e, f). g Quantification of nerve length of abducens, (h) oculomotor, and (i) trochlear nerves. Kruskal-Wallis test followed by the Steel-Dwass test for the abducens and oculomotor nerves, one-way ANOVA for the trochlear nerve, *p < 0.05, **p < 0.01. j–o The three axon guidance patterns in E12.5 homozygous mutant abducens nerves. Substantial numbers of wandering nerves (arrows) were present in homozygous mutant abducens nerves (l, m). The wandering phenotype was defined as abducens nerves with inappropriately extending nerves of substantial length (> 30% of appropriately innervated nerves). The edge (*) of mutant abducens nerves was far from the target area (n, o). The stalled phenotype was defined as abducens nerves not reaching the target area. p The ratio of abducens nerves with abnormal axon guidance. Scale bar: 500 μm (a–c), 200 μm (d–f, j–o). Abbreviations: III, oculomotor nerve; IV, trochlear nerve; VI, abducens nerve; XII, hypoglossal nerve