MZrgs4 embryos show a specific decrease in the number of HuC+ cells and a significant increase in the number of PH3+ cells in the spinal cord. (A) Lateral views of spinal cord Z projections (around 15 μm) following whole-mount immunostaining for HuC (left) and counterstained with DAPI (middle) with merged still images (right) in WT (top) and MZrgs4 larvae (bottom) at 48 hpf. Scale bar = 20 μm. (B) Quantification of the number of HuC+ and HuC− cells within the spinal cord in WT (average of 101.2 ± 3.12 for HuC+ ; average of 93.83 ± 7.36 for HuC−, n = 6) and MZrgs4 larvae (average of 51.86 ± 5.25 for HuC+ ; average of 84.71 ± 4.39 for HuC−, n = 7) at 35 hpf (left) and at 48 hpf in WT (right) (average of 166.7 ± 7.04 for HuC+ ; average of 146.2 ± 8.28 for HuC−, n = 6) and MZrgs4 larvae (average of 130.3 ± 3.7 for HuC+ ; average of 153.1 ± 3.99 for HuC−, n = 7). (C) Lateral views of spinal cord Z projections (around 30 μm for PH3) following whole-mount immunostaining for PH3 (left) and HuC (middle) with merged still images (right) in WT (top) and MZrgs4 larvae (bottom) at 48 hpf. (D) Quantification of the number of PH3+ cells within the spinal cord in WT (average of 14.29 ± 1.82 at 35 hpf, n = 7; average of 4 ± 1.08 at 48 hpf, n = 9) and MZrgs4 larvae (average of 19.56 ± 1.10 at 35 hpf, n = 9; average of 12.44 ± 1.51 at 48 hpf, n = 9). (E) Dorsal views of head Z projections (around 400 μm) following HuC whole-mount immunostaining in WT and MZrgs4 larvae at 48 hpf. Dashed lines represent the nervous system. Scale bar = 50 μm. (F) Quantification of the head area in WT (average of 180,194 ± 3289, n = 9) and MZrgs4 larvae (average of 153,316 ± 5151, n = 9) at 48 hpf. (G) Quantification of the head perimeter in WT (average of 1677 ± 31.90, n = 9) and MZrgs4 larvae (average of 1564 ± 30.24, n = 9) at 48 hpf.
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