RNA expression in anterior nasal septum cartilage of wildtype and Fgfr2+/S252W Apert syndrome model mice at embryonic day (E)14.5.

The nasal capsule cartilage of Apert Fgfr2+/S252W mice increases in size compared to wildtype cartilage during embryonic craniofacial development. Increased levels of markers of cell proliferation can be detected in the Apert Fgfr2+/S252W anterior nasal septum cartilage compared to wildtype at E14.5. We have performed laser capture microdissection of this cartilage in these genotypes at this age to generate RNA-Seq libraries and compare gene expression. Expression analysis shows increased expression of genes related to chromosome condensation, nuclear division, and the cell cycle in Apert Fgfr2+/S252W anterior nasal septum cartilage compared to wildtype. Laser capture microdissection was performed on fresh frozen sections of three wildtype and three mutant mice. RNA was isolated from the six individual samples, converted to RNA-Seq libraries, and sequenced on the Illumina HiSeq 2500 platform.Apert Fgfr2+/S252W 小鼠的鼻囊软骨在胚胎颅面发育过程中，其体积较野生型（wildtype）软骨有所增大。在胚胎发育第14.5天（E14.5），Apert Fgfr2+/S252W小鼠的前鼻中隔软骨中可检测到的细胞增殖标志物水平较野生型更高。我们针对该日龄的两种基因型小鼠的该类软骨开展了激光捕获显微切割（laser capture microdissection）实验，以构建RNA测序（RNA-Seq）文库并比较基因表达差异。表达分析结果显示，相较于野生型，Apert Fgfr2+/S252W小鼠的前鼻中隔软骨中，与染色体凝缩、核分裂及细胞周期相关的基因表达水平显著升高。本实验对3只野生型与3只突变型小鼠的新鲜冰冻切片开展了激光捕获显微切割。从这6份独立样本中提取RNA，构建RNA-Seq文库，并在Illumina HiSeq 2500测序平台上完成测序。