Massa Luciana F, Ramachandran Amsaveni, George Anne, Arana-Chavez Victor E
Laboratory of Mineralized Tissue Biology, Department of Cell and Developmental Biology, Institute of Biomedical Sciences, University of São Paulo, 05508-900, São Paulo, Brazil.
Histochem Cell Biol. 2005 Sep;124(3-4):197-205. doi: 10.1007/s00418-005-0009-9. Epub 2005 Oct 28.
Dentin matrix protein 1 (DMP 1) is an acidic phosphoprotein that has been postulated to play an important role in mineralized tissue formation. We have examined rat molar tooth germs by applying a high-resolution immunocytochemical approach with the purpose to identify the temporal and spatial localization of DMP 1 at the onset of dentinogenesis. Upper molar tooth germs of 2- to 3-day-old Wistar rats were fixed in a cacodylate-buffered 0.1% glutaraldehyde + 4% formaldehyde fixative, left unosmicated and embedded in LR White resin. The sections were incubated with a polyclonal DMP 1 antibody for postembedding colloidal gold immunolabeling and examined in a Jeol 1010 transmission electron microscope. The earliest localization of DMP 1 was in the Golgi region as well as in the nucleus of differentiating odontoblasts. When mineralization spread from matrix vesicles to the surrounding matrix, DMP 1 was extracellularly detected around the mineralizing globules. In the regions of fully mineralized mantle dentin, it was present in the mineralized regions, mainly around the peritubular dentin. The appearance of DMP 1 during early dentinogenesis implies a direct role for this protein in both odontoblast differentiation and matrix mineralization.
牙本质基质蛋白1(DMP 1)是一种酸性磷酸蛋白,据推测在矿化组织形成中起重要作用。我们应用高分辨率免疫细胞化学方法检查了大鼠磨牙牙胚,目的是确定牙本质形成开始时DMP 1的时空定位。将2至3日龄Wistar大鼠的上颌磨牙牙胚固定在二甲胂酸盐缓冲的0.1%戊二醛+4%甲醛固定剂中,不进行锇化处理,然后包埋在LR White树脂中。切片用多克隆DMP 1抗体孵育,进行包埋后胶体金免疫标记,并在Jeol 1010透射电子显微镜下检查。DMP 1最早的定位是在高尔基体区域以及分化中的成牙本质细胞的细胞核中。当矿化从基质小泡扩散到周围基质时,在矿化小球周围细胞外检测到DMP 1。在完全矿化的罩牙本质区域,它存在于矿化区域,主要围绕管周牙本质。DMP 1在早期牙本质形成过程中的出现意味着该蛋白在成牙本质细胞分化和基质矿化中都起直接作用。