Houari Sophia, Babajko Sylvie, Loiodice Sophia, Berdal Ariane, Jedeon Katia
Institut National de la Santé et Recherche Médicale (INSERM) UMRS 1138, Paris-Diderot University, Pierre & Marie Curie University, Paris-Descartes University, Laboratory of Molecular Oral Pathophysiology, Cordeliers Research Centre; Unit of Formation and Research (UFR) of Odontology, Paris-Diderot University.
Institut National de la Santé et Recherche Médicale (INSERM) UMRS 1138, Paris-Diderot University, Pierre & Marie Curie University, Paris-Descartes University, Laboratory of Molecular Oral Pathophysiology, Cordeliers Research Centre; Unit of Formation and Research (UFR) of Odontology, Paris-Diderot University;
J Vis Exp. 2018 Mar 29(133):57081. doi: 10.3791/57081.
Enamel defects resulting from environmental conditions and ways of life are public health concerns because of their high prevalence. These defects result from altered activity of cells responsible for enamel synthesis named ameloblasts, which present in enamel organ. During amelogenesis, ameloblasts follow a specific and precise sequence of events of proliferation, differentiation, and death. A rat continually growing incisors is a suitable experimental model to study ameloblast activity and differentiation stages in physiological and pathological conditions. Here, we describe a reliable and consistent method to micro-dissect enamel organ of rats exposed to environmental toxicants. The micro-dissected dental epithelia contain secretion- and maturation-stage ameloblasts that may be used for qualitative experiments, such as immunohistochemistry assays and in situ hybridization, as well as for quantitative analyses such as RT-qPCR, RNA-seq, and Western blotting.
由于环境条件和生活方式导致的牙釉质缺陷因其高患病率而成为公共卫生问题。这些缺陷是由负责牙釉质合成的细胞（即成釉细胞）活性改变所致，成釉细胞存在于牙釉质器官中。在牙釉质形成过程中，成釉细胞遵循特定且精确的增殖、分化和死亡事件序列。大鼠不断生长的切牙是研究生理和病理条件下成釉细胞活性及分化阶段的合适实验模型。在此，我们描述一种可靠且一致的方法，用于显微解剖暴露于环境毒物的大鼠牙釉质器官。显微解剖的牙上皮包含分泌期和成熟期的成釉细胞，可用于定性实验，如免疫组织化学分析和原位杂交，也可用于定量分析，如RT-qPCR、RNA测序和蛋白质免疫印迹。
