|
Fig. 2 sec24d mRNA expression during embryogenesis and molecular analysis of the bulldog mutation.
(A) RT-PCR assay shows maternally deposited sec24d transcripts at 1 and 2 hpf and increasing levels thereafter. β-actin served as control. (B–H) Expression patterns of sec24d during zebrafish development using whole mount in situ hybridization staining of wild-type embryos. The cross section through the palatoquadrate (pq, arrowheads) at 3 dpf (I) shows robust riboprobe staining (h: hours, d: days). (J–O) Expression patterns of col2α1 (J,K) and sox9a (L,M) at 3 dpf. The expression is slightly upregulated in the posterior pharyngeal cartilages of bulm606 embryos. The Collagen-specific chaperone hsp47 (N,O) transcript is elevated in craniofacial skeleton (arrowheads), fins and the notochord. Arrowheads point to the jaw region in panels (G) through (O). (P) Relative sec24d sec24c, bip and sil1 mRNA levels were assessed by quantitative real-time PCR and adjusted against β-actin using total RNA samples from 4 dpf embryos. In bulm494, mRNA levels for bip and sil1 are induced 5-fold, whereas sec24d mRNA levels are reduced 0.7-fold. *, p<0.02; **, p<0.005. Abbreviation: b, brain, o, optic nerve, ot, otic capsule, r, retina. Anterior (D–O) is to the left.