Fig. 3

Fig. 3

Depletion and overexpression of Ostn result in shortening and elongation, respectively, of the parasphenoid at an early stage. (A) DNA and corresponding amino acid sequence of the ostn^ncv105 allele. The 5 bp deletion in ostn^ncv105 leads to the mutation of 40 amino acids followed by a premature termination (asterisk). (B) Genotype of wild-type (+), heterozygous (+/ncv105) and homozygous (ncv105) alleles as confirmed by digestion of PCR products containing a PstI site as indicated in A. (C) Ratio of head length [from the tip of the upper jaw to the caudal part of the Alizarin Red stained notochord (nt)] to body length (from the tip of the upper jaw to the end of the tail) of 7 dpf wild-type (+) or homozygous mutant (ncv105) larvae. Each point indicates data from an individual larva. n, number of larvae examined. (D) Representative images of Alizarin Red staining of 7 dpf larvae of wild type (top), ostn^ncv105 homozygous mutant (middle) and Tg(myl7:ostn,hsp70l:EGFP) (bottom). Lateral view (left) and ventral view (right), anterior to the left. Arrows and dashed lines indicate the tip and the top of the concave in the caudal part of the parasphenoid (ps), respectively. (E) Quantitative analyses of cranial bone length of 7 dpf ostn^ncv105 and transgenic larvae relative to the mean length of wild-type bones is plotted for entopterygoid (ent), ps (between arrow and dashed line in D), branchiostegal ray (bsr) and cleithrum (cl). (F) Representative images of Alizarin Red staining of 7 dpf larvae of wild type (top), ostn^ncv105 homozygous mutant (middle) and Tg(myl7:ostn,hsp70l:EGFP);ostn^ncv105. Views as in D. (G) Quantitative analyses of F. Data were analyzed by Student's t-test or one-way ANOVA with Tukey's test. *P<0.05. Error bars indicate s.e.m. Scale bars: 200 µm.