Sigal M J, Aubin J E, Ten Cate A R
J Dent Res. 1985 Dec;64(12):1348-55. doi: 10.1177/00220345850640120401.
An immunofluorescence technique was applied at the light microscope level to human third molar coronal dentin in order to localize the intracellular components tubulin, vimentin, and actin. Third molars were split immediately upon extraction, and immersed in periodate-lysine-paraformaldehyde fixative. The crowns were demineralized, dehydrated, and wax-embedded, and 6-micron sections were prepared. The sections were post-fixed in -20 degrees C acetone, and then incubated with monoclonal mouse anti-tubulin, anti-vimentin, or anti-actin antibodies, followed by fluorescein-conjugated sheep anti-mouse immunoglobulins. Intratubular immunofluorescence labeling for tubulin and vimentin was very similar in pattern and intensity and extended to the dentino-enamel junction. In contrast, the actin labeling appeared less intense and more punctate, and was located primarily in the pulpal half of the crown, although some labeling was detectable up to the dentino-enamel junction. The presence of tubulin-, vimentin-, and actin-containing structures extending to the dentino-enamel junction supports the hypothesis that the odontoblast process does extend to the dentino-enamel junction in the human, and is in agreement with earlier studies of rat molars.
为了定位细胞内成分微管蛋白、波形蛋白和肌动蛋白，在光学显微镜水平上对人类第三磨牙的冠部牙本质应用了免疫荧光技术。第三磨牙在拔除后立即劈开，并浸入高碘酸盐-赖氨酸-多聚甲醛固定剂中。牙冠进行脱矿、脱水和石蜡包埋，然后制备6微米厚的切片。切片在-20℃丙酮中后固定，然后与小鼠抗微管蛋白、抗波形蛋白或抗肌动蛋白单克隆抗体孵育，随后用荧光素偶联的羊抗小鼠免疫球蛋白处理。微管蛋白和波形蛋白的管内免疫荧光标记在模式和强度上非常相似，并延伸至牙本质-釉质界。相比之下，肌动蛋白标记显得强度较低且更呈点状，主要位于牙冠的牙髓侧，尽管在牙本质-釉质界处也可检测到一些标记。含有微管蛋白、波形蛋白和肌动蛋白的结构延伸至牙本质-釉质界，这支持了成牙本质细胞突起确实延伸至人类牙本质-釉质界的假说，并且与早期对大鼠磨牙的研究结果一致。
