IMMUNOHISTOCHEMICAL DEMONSTRATION OF AMELOGENIN PENETRATION TOWARD THE DENTAL-PULP IN THE EARLY STAGES OF AMELOBLAST DEVELOPMENT IN RAT MOLAR TOOTH GERMS
IMMUNOHISTOCHEMICAL DEMONSTRATION OF AMELOGENIN PENETRATION TOWARD THE DENTAL-PULP IN THE EARLY STAGES OF AMELOBLAST DEVELOPMENT IN RAT MOLAR TOOTH GERMS
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DOI：
10.1002/ar.1092290213
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发表时间：
1991-02-01
影响因子：
--
通讯作者：
KURISU, K
中科院分区：
文献类型：
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作者：
INAI, T;KUKITA, T;KURISU, K
为了研究成釉细胞在发育早期合成和分泌成釉蛋白的情况，用初步实验中制备的单抗在光镜和电子显微镜水平上进行了免疫组织化学定位。材料为出生后0~5d的大鼠下颌第一磨牙牙胚。双向电泳免疫印迹分析表明，该单抗识别的抗原分子为26-28 kDa(等电点为6.6-7.0)的成釉蛋白。免疫组织化学检测表明，发育早期的成釉细胞既合成了釉原蛋白，又通过经典的内分泌分泌途径分泌。在一些成熟前成釉细胞和成釉细胞中，我们观察到粗面内质网(RER)池的膨大，这是一种异质免疫标记。在前牙本质以及成牙本质细胞和牙髓细胞的细胞间隙中也检测到免疫标记，表明成釉蛋白从记忆前成釉细胞层渗透到牙髓中。成牙本质细胞质膜上靠近釉质蛋白的包被凹坑的存在，以及成牙本质细胞溶酶体的免疫标记，表明成牙本质细胞吞噬了釉质蛋白。这些观察表明，釉质蛋白对牙髓和成牙本质细胞的渗透可能在成釉细胞和成牙本质细胞之间的相互作用中发挥作用。
In order to examine the synthesis and secretion of enamel protein by ameloblasts in their early stages of development, immunohistochemical localization was carried out at light and electron microscopic levels using a monoclonal antibody produced in a preliminary experiment. Materials used were tooth germs of mandibular first molars of rats at 0-5 days after birth. Immunoblot analysis after two-dimensional electrophoresis revealed that antigen molecules recognized by the monoclonal antibody were amelogenins of 26-28 kDa (pI, 6.6-7.0).An immunohistochemical examination using this monoclonal antibody demonstrated that the presecretory ameloblasts in their early stages of differentiation both synthesized amelogenin and secreted through a classical merocrine secretory pathway. In some presecretory ameloblasts as well as ameloblasts we observed the distended cisternae of rough endoplasmic reticulum (rER) which demonstrated heterogenous immunolabelling. The immunolabellings were also detected in the predentin as well as the intercellular spaces of odontoblasts and dental pulp cells which indicated penetration of amelogenin from the presecretory ameloblast layer to the dental pulp. The presence of coated pits at the plasma membrane of odontoblasts in close proximity to enamel protein along with the immunolabelling of lysosomes of the odontoblasts suggests the phagocytosis of the enamel protein into the odontoblasts. These observations suggest the possibility that the penetration of enamel protein toward the dental pulp and odontoblasts plays a role in the interaction between ameloblasts and odontoblasts.