Gebuijs I G E, Raterman S T, Metz J R, Swanenberg L, Zethof J, Van den Bos R, Carels C E L, Wagener F A D T G, Von den Hoff J W
Department of Orthodontics and Craniofacial Biology, Radboudumc, Nijmegen, The Netherlands.
Department of Orthodontics and Craniofacial Biology, Radboud Institute of Molecular Life Sciences, Nijmegen, The Netherlands.
Biol Open. 2019 Sep 9;8(9):bio039834. doi: 10.1242/bio.039834.
Craniofacial development is tightly regulated and therefore highly vulnerable to disturbance by genetic and environmental factors. Fibroblast growth factors (FGFs) direct migration, proliferation and survival of cranial neural crest cells (CNCCs) forming the human face. In this study, we analyzed bone and cartilage formation in the head of five dpf zebrafish larvae and assessed gene expression levels for 11 genes involved in these processes. In addition, hybridization was performed on 8 and 24 hours post fertilization (hpf) larvae (, , , ). A significant size reduction of eight out of nine craniofacial cartilage structures was found in homozygous mutant (6-36%, <0.01) and heterozygous (7-24%, <0.01) larvae. Also, nine mineralized structures were not observed in all or part of the homozygous (0-71%, <0.0001) and heterozygous (33-100%, <0.0001) larvae. In homozygote mutants, and expression was upregulated compared to wild type, presumably to compensate for the reduced bone formation. Decreased expression may compromise cartilage formation. Upregulated in homozygotes indicates impaired CNCC function. expression was reduced in the first and second stream of CNCCs in homozygous mutants at 24 hpf, as shown by hybridization. This indicates an impairment of CNCC migration and survival by mutation.
颅面发育受到严格调控，因此极易受到遗传和环境因素的干扰。成纤维细胞生长因子（FGFs）指导形成人类面部的颅神经嵴细胞（CNCCs）的迁移、增殖和存活。在本研究中，我们分析了5天龄斑马鱼幼虫头部的骨和软骨形成，并评估了参与这些过程的11个基因的基因表达水平。此外，还对受精后8小时和24小时的幼虫进行了杂交（，，，）。在纯合突变体（6 - 36%，<0.01）和杂合子（7 - 24%，<0.01）幼虫中，发现九个颅面软骨结构中的八个有显著尺寸减小。同样，在全部或部分纯合子（0 - 71%，<0.0001）和杂合子（33 - 100%，<0.0001）幼虫中未观察到九个矿化结构。在纯合突变体中，与野生型相比，和表达上调，推测是为了补偿骨形成的减少。表达降低可能会损害软骨形成。纯合子中上调表明CNCC功能受损。如杂交所示，在24小时时，纯合突变体中CNCC的第一和第二流中的表达降低。这表明突变会损害CNCC的迁移和存活。
