Lesot H, Karcher-Djuricic V, Kubler M D, Ruch J V
Institut de Biologie Médicale, CNRS LP 6520, Faculté de Médecine, Strasbourg, France.
Differentiation. 1988;37(1):62-72. doi: 10.1111/j.1432-0436.1988.tb00797.x.
It is known that high-molecular-weight (HMW) membrane proteins mediate interactions with constituents of the extracellular matrix and/or with cytoskeletal elements. To study participation of HMW membrane proteins in odontoblast or ameloblast differentiation, an immunological approach has been adopted. Antibodies directed against membrane proteins (Mr, 110-190) from mouse embryos have been produced by the hybridoma technique. Supernatants of hybridoma cultures were screened for their ability to stain dental tissues and also tested for their biological activities on dental cells in primary culture or on developing tooth germs in organ culture. An IgM monoclonal antibody, MC16A16, directed against a 165-kDa antigen present in plasma membrane preparations, reacted strongly with the dental epithelium and weakly with the mesenchyme. MC16A16 also reacted with the cell surface of nonpermeabilized cultured dental cells and could detach epithelial cells cultured on glass, but not mesenchymal cells which maintained vinculin-containing focal contacts. This antibody, which affected the organization of dental-cell microfilaments in primary culture, also inhibited the polarization of odontoblasts, but not that of ameloblasts.
已知高分子量(HMW)膜蛋白介导与细胞外基质成分和/或细胞骨架成分的相互作用。为了研究HMW膜蛋白在成牙本质细胞或成釉细胞分化中的作用,采用了一种免疫学方法。通过杂交瘤技术制备了针对小鼠胚胎膜蛋白(分子量为110 - 190)的抗体。对杂交瘤培养物的上清液进行筛选,以检测其对牙组织的染色能力,并测试其对原代培养的牙细胞或器官培养中正在发育的牙胚的生物学活性。一种IgM单克隆抗体MC16A16,针对存在于质膜制剂中的165 kDa抗原,与牙上皮强烈反应,与间充质反应较弱。MC16A16还与未通透的培养牙细胞的细胞表面反应,并能使培养在玻璃上的上皮细胞脱离,但不能使维持含纽蛋白粘着斑的间充质细胞脱离。这种抗体在原代培养中影响牙细胞微丝的组织,也抑制成牙本质细胞的极化,但不抑制成釉细胞的极化。