Identification of stages of amelogenesis in the continuously growing mandiblular incisor of C57BL/6J male mice throughout life using molar teeth as landmarks.
Identification of stages of amelogenesis in the continuously growing mandiblular incisor of C57BL/6J male mice throughout life using molar teeth as landmarks.
复制标题
DOI：
10.3389/fphys.2023.1144712
复制
发表时间：
2023
影响因子：
4
通讯作者：
中科院分区：
文献类型：
--
作者：
连续生长的小鼠切牙被广泛用于研究釉质发生，因为该过程的所有阶段（即，分泌、转变和成熟）在任何给定时间以空间上确定的顺序存在。为了研究与釉质形成相关的生物学变化，重要的是开发可靠的方法来收集成釉细胞，从釉质形成的不同阶段调节釉质形成的细胞。显微解剖是从小鼠切牙中收集不同成釉细胞群体的关键方法，它依赖于磨牙的位置作为识别成釉关键阶段的标志。然而，下颌切牙的位置及其与磨牙的空间关系随着年龄的变化而变化。我们的目标是高精度地确定这些关系在整个骨骼生长和老年人，哺乳动物成熟。收集2、4、8、12、16、24周龄和18月龄C57 BL/6 J雄性小鼠的下颌骨，并使用显微CT和组织学进行研究，以获得切牙釉质矿化概况，并确定成釉细胞形态在成釉过程中相对于磨牙位置的相应变化。正如本文所报道的，我们发现在整个活跃的骨骼生长过程中（2-16周），切牙的根尖和釉质矿化的开始相对于磨牙向远侧移动。过渡阶段的位置也向远侧移动。为了验证标记的准确性，我们将12周龄动物下颌切牙的釉质上皮显微解剖成5个段，包括1）分泌期，2）晚期分泌-过渡-早期成熟，3）早期成熟，4）中期成熟和5）晚期成熟。合并分离的片段，并使用RT-qPCR对编码关键釉基质蛋白（EMP）、Amelx、Enam和Odam的基因进行表达分析。Amelx和Enam在分泌期强烈表达（第1段），而它们的表达在过渡期减少（第2段），并在成熟期停止（第3、4和5段）。相比之下，Odam的表达在分泌过程中非常低，并在整个过渡和成熟阶段急剧增加。这些表达谱与釉基质蛋白表达的共识一致。总体而言，我们的研究结果表明，我们的地标方法的准确性很高，并强调选择年龄合适的标志在小鼠切牙的釉质发生的研究的重要性。
Continuously growing mouse incisors are widely used to study amelogenesis, since all stages of this process (i.e., secretory, transition and maturation) are present in a spatially determined sequence at any given time. To study biological changes associated with enamel formation, it is important to develop reliable methods for collecting ameloblasts, the cells that regulate enamel formation, from different stages of amelogenesis. Micro-dissection, the key method for collecting distinct ameloblast populations from mouse incisors, relies on positions of molar teeth as landmarks for identifying critical stages of amelogenesis. However, the positions of mandibular incisors and their spatial relationships with molars change with age. Our goal was to identify with high precision these relationships throughout skeletal growth and in older, skeletally mature animals. Mandibles from 2, 4, 8, 12, 16, and 24-week-old, and 18-month-old C57BL/6J male mice, were collected and studied using micro-CT and histology to obtain incisal enamel mineralization profiles and to identify corresponding changes in ameloblast morphology during amelogenesis with respect to positions of molars. As reported here, we have found that throughout active skeletal growth (weeks 2–16) the apices of incisors and the onset of enamel mineralization move distally relative to molar teeth. The position of the transition stage also moves distally. To test the accuracy of the landmarks, we micro-dissected enamel epithelium from mandibular incisors of 12-week-old animals into five segments, including 1) secretory, 2) late secretory - transition - early maturation, 3) early maturation, 4) mid-maturation and 5) late maturation. Isolated segments were pooled and subjected to expression analyses of genes encoding key enamel matrix proteins (EMPs), Amelx, Enam, and Odam, using RT-qPCR. Amelx and Enam were strongly expressed during the secretory stage (segment 1), while their expression diminished during transition (segment 2) and ceased in maturation (segments 3, 4, and 5). In contrast, Odam’s expression was very low during secretion and increased dramatically throughout transition and maturation stages. These expression profiles are consistent with the consensus understanding of enamel matrix proteins expression. Overall, our results demonstrate the high accuracy of our landmarking method and emphasize the importance of selecting age-appropriate landmarks for studies of amelogenesis in mouse incisors.
登录
查看更多内容
影响因子：
4.6
作者：
Liang, Tian;Wang, Shih-Kai;Smith, Charles;Zhang, Hong;Hu, Yuanyuan;Seymen, Figen;Koruyucu, Mine;Kasimoglu, Yelda;Kim, Jung-Wook;Zhang, Chuhua;Saunders, Thomas L.;Simmer, James P.;Hu, Jan C. -C.
通讯作者：
Hu, Jan C. -C.
影响因子：
3
作者：
Gallon, Victoria;Chen, Lisha;Yang, Xiudong;Moradian-Oldak, Janet
通讯作者：
Moradian-Oldak, Janet
影响因子：
4.6
作者：
Farooq S;Leussink S;Sparrow LM;Marchini M;Britz HM;Manske SL;Rolian C
通讯作者：
Rolian C
影响因子：
5.6
作者：
Lacruz, Rodrigo S.;Smith, Charles E.;Moffatt, Pierre;Chang, Eugene H.;Bromage, Timothy G.;Bringas, Pablo, Jr.;Nanci, Antonio;Baniwal, Sanjeev K.;Zabner, Joseph;Welsh, Michael J.;Kurtz, Ira;Paine, Michael L.
通讯作者：
Paine, Michael L.
通过全基因组转录谱分析鉴定参与牙釉质矿化的新候选基因。
影响因子：
5.6
作者：
Lacruz, Rodrigo S.;Smith, Charles E.;Bringas, Pablo, Jr.;Chen, Yi-Bu;Smith, Susan M.;Snead, Malcolm L.;Kurtz, Ira;Hacia, Joseph G.;Hubbard, Michael J.;Paine, Michael L.
通讯作者：
Paine, Michael L.