Activated Rap1A Induces Osteoblastic Differentiation and Cell Adhesion

다국어 초록 (Multilingual Abstract)

Rap1 is a key regulator of cell adhesion and migration. Although increasing evidence indicates that the Rap1 signaling pathway is involved in the process of bone remodeling, the mechanism by which Rap1 regulates osteoblastic differentiation and cell adhesion remains unknown. Here, we investigated the morphological characteristics and osteoblastic differentiation of cells expressing constitutively activated form of Rap1A (Rap1ACA) or Rap1 GTPase activating protein Rap1GAP and found that activated Rap1 induces osteoblastic differentiation and cell adhesion as well as cell spreading. When osteoblastic differentiation was induced, Rap1ACA cells showed considerably higher levels of calcium deposits than the wild-type and Rap1GAP-overexpressing cells did. Rap1ACA cells showed increased spreading and size, as well as strong cell adhesion and significantly decreased growth rates. F-actin staining using phalloidin revealed several thin thread-like filopodia around the protrusions in Rap1ACA cells, which possibly contribute to the increased cell adhesion.

목차 (Table of Contents)

• 1. Introduction
• 2. Materials and Methods
• 2.1. Materials and Plasmids
• 2.2. Cell Culture and Stable Cell Lines
• 2.3. Osteogenic Induction
• 2.4. Cell Adhesion Assay
• 2.5. Cell Growth Rate Assay
• 2.6. Immunofluorescence
• 2.7. Alizarin Red S Staining
• 2.8. Statistical Analysis
• 3. Results and Discussion
• 3.1. Activated Rap1A Induces Osteoblastic Differentiation in Preosteoblasts
• 3.2. Activated Rap1A Stimulates Spreading of Preosteoblasts
• 3.3. Activated Rap1A Promotes Formation of Filopodia and Increases Cell Adhesion
• 4. Conclusion
• References