Colocalization of TGF-beta 1 and extracellular matrix proteins during rat tooth development.

R N D'Souza, K Flanders, W T Butler
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Abstract

Recent studies suggest that TGF-beta 1 controls events in embryogenesis via its modulation of the extracellular matrix (ECM). During tooth development, histomorphogenetic and cytodifferentiation events are regulated by ECM-mediated epithelial-mesenchymal interactions. In this study, we have investigated the in vivo relationship of the localization of TGF-beta 1 to various ECM proteins, including glycosaminoglycans (GAGs), fibronectin, collagens I and III and to the enzyme alkaline phosphatase, at the early and late bell stages of rat molar tooth development. We have used polyclonal antibodies specific to peptides present in the precursor and processed forms of TGF-beta 1, fibronectin and Collagens I and III and immunohistochemical methods of detection. GAGs were stained with Alcian Blue GX and alkaline phosphatase was detected with the enzyme Naphthol-AS B1. Our results indicate that both the intracellular and extracellular forms of TGF-beta 1 are expressed in a site-specific manner, and that the growth factor colocalizes with alkaline phosphatase, fibronectin, collagens I and III and GAGs during the bell stages of tooth development. These data support the hypothesis that in tooth development, the actions of TGF-beta 1 may contribute to important qualitative changes in the composition and structure of extracellular matrices.

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tgf - β 1和细胞外基质蛋白在大鼠牙齿发育过程中的共定位。
最近的研究表明,tgf - β 1通过调节细胞外基质(ECM)来控制胚胎发生的事件。在牙齿发育过程中,组织形态发生和细胞分化事件是由ecm介导的上皮-间质相互作用调节的。在本研究中,我们研究了tgf - β 1在大鼠磨牙发育早期和晚期与多种ECM蛋白(包括糖胺聚糖(GAGs)、纤维连接蛋白、胶原I和III以及碱性磷酸酶)定位的体内关系。我们使用了针对tgf - β 1前体和加工形式中存在的肽的多克隆抗体,纤维连接蛋白和胶原I和III以及免疫组织化学检测方法。用Alcian Blue GX染色GAGs,用萘酚- as B1酶检测碱性磷酸酶。我们的研究结果表明,细胞内和细胞外形式的tgf - β 1都以位点特异性的方式表达,并且在牙齿发育的钟形阶段,生长因子与碱性磷酸酶、纤维连接蛋白、胶原I和III以及gag共定位。这些数据支持了一种假设,即在牙齿发育过程中,tgf - β 1的作用可能有助于细胞外基质组成和结构的重要质的变化。
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