Fig. 2

Fig. 2

gon4la mutants show impaired growth and delayed craniofacial development. (A) Reduced body size and (B) significantly smaller total bone and skull volume in gon4laⁿⁿ²¹¹² mutants (3 mpf, n = 5/group). (C) Survival proportions of gon4laⁿⁿ²¹¹² homozygotes begin to decline after 12 wpf (Genotyping n ≥ 96/time point). (D, E) Growth metrics (length and weight) show delayed growth post-8 wpf (n ranges from 10/4 at 1 wpf to 11/6 at 16 wpf for WT/homo). (F) Comparison of body mass index (BMI) across all three mutant lines, showing a systemic reduction in body mass relative to length. (G, H) Craniofacial bone structure of WT siblings and gon4laⁿⁿ²¹¹² mutant larvae stained with Alcian blue at 5 dpf. (I) Quantification of six distinct length measurements (a–f), including total head length, ceratohyal cartilage length, and various joint distances, as indicated in (G). (J) Quantification of four key craniofacial cartilage angles: ceratohyal cartilages (CH/CH), palatoquadrate and ceratohyal (PQ/CH), Meckel’s cartilages (MCK/MCK), and palatoquadrate and Meckel’s cartilage (PQ/MCK) (n = 9–10/group). (K) µCT images of 3–4 mpf skulls (lateral view): Opercle (op, blue) and metapterygoid (mpt, red) are segmented and labeled. (L) Representative lateral and ventral views of segmented op and mpt from WT siblings and gon4laⁿⁿ²¹¹² mutants. (M) The sizes of op and mpt and their ratios to skull size were analyzed in all three mutant lines (n = 5–6/group).