Supplementary Material for: Phenotype-Genotype Correlations in Mouse Models of<b> </b>Amelogenesis Imperfecta<b> </b>Caused by<b><i> Amelx </i></b>and<b><i> Enam </i></b>Mutations

Mutations in human and in mouse orthologous genes <i>Amelx</i> and <i>Enam </i>result in a diverse range of enamel defects. In this study we aimed to investigate the phenotype-genotype correlation between the mutants and the wild-type controls in mouse models of amelogenesis imperfecta using novel measurement approaches. Ten hemi-mandibles and incisors were dissected from each group of <i>Amelx</i>^WT, <i>Amelx</i>^X/Y64H, <i>Amelx</i>^Y/Y64H, <i>Amelx</i>^Y64H/Y64H, and<i> Enam</i>^WT, <i>Enam</i>^Rgsc395 heterozygous and <i>Enam</i>^Rgsc395 homozygous mice. Their macro-morphology, colour and micro-topography were assessed using bespoke 2D and 3D image analysis systems and customized colour and whiteness algorithms. The novel methods identified significant differences (p ≤ 0.05) between the <i>Amelx</i> groups for mandible and incisor size and enamel colour and between the <i>Enam</i> groups for incisor size and enamel colour. The <i>Amelx</i>^WT mice had the largest mandibles and incisors, followed in descending order of size by the <i>Amelx</i>^X/Y64H, <i>Amelx</i>^Y/Y64H and <i>Amelx</i>^Y64H/Y64H mice. Within the <i>Enam</i> groups the <i>Enam</i>^WT incisors were largest and the <i>Enam</i>^Rgsc395 heterozygous mice were smallest. The effect on tooth morphology was also reflected by the severity of the enamel defects in the colour and whiteness assessment. Amelogenin affected mandible morphology and incisor enamel formation, while enamelin only affected incisors, supporting the multifunctional role of amelogenin. The enamelin mutation was associated with earlier forming enamel defects. The study supported the critical involvement of amelogenin and enamelin in enamel mineralization.人类与小鼠的同源基因<em>Amelx</em>及<em>Enam</em>发生突变后，可引发多样化的牙釉质缺陷。本研究采用新型检测方法，针对牙釉质发育不全（amelogenesis imperfecta）小鼠模型，探究突变体与野生型对照之间的表型-基因型关联。本研究从<em>Amelx</em>^{野生型（WT）}、<em>Amelx</em>^X/Y64H、<em>Amelx</em>^Y/Y64H、<em>Amelx</em>^Y64H/Y64H组，以及<em>Enam</em>^{野生型（WT）}、<em>Enam</em>^Rgsc395杂合子与纯合子小鼠的各实验组中，各分离获得10个半下颌骨与切牙。研究人员通过定制化的二维与三维图像分析系统，以及定制化的颜色与白度算法，对样本的大体形态、色泽与微观形貌进行评估。新型检测方法显示，<em>Amelx</em>组的下颌骨与切牙尺寸、牙釉质色泽，以及<em>Enam</em>组的切牙尺寸与牙釉质色泽均存在显著差异（p ≤ 0.05）。<em>Amelx</em>^{野生型（WT）}小鼠的下颌骨与切牙尺寸最大，其后按尺寸降序依次为<em>Amelx</em>^X/Y64H、<em>Amelx</em>^Y/Y64H与<em>Amelx</em>^Y64H/Y64H小鼠。在<em>Enam</em>组中，<em>Enam</em>^{野生型（WT）}小鼠的切牙尺寸最大，而<em>Enam</em>^Rgsc395杂合子小鼠的切牙尺寸最小。牙齿形态所受的影响，也可通过色泽与白度评估中的牙釉质缺陷严重程度得以体现。成釉蛋白可影响下颌骨形态与切牙牙釉质形成，而釉蛋白仅对切牙产生影响，这支持了成釉蛋白的多功能性作用。釉蛋白突变与更早出现的牙釉质缺陷相关。本研究证实了成釉蛋白与釉蛋白在牙釉质矿化过程中的关键作用。