Ephrin-B1 reverse signaling in craniofacial development

描述(申请人提供)：在颅面发育过程中控制神经脊迁移的机制还不是很清楚。Eph细胞信号受体-配体对在发育过程中起着至关重要的作用，包括控制神经脊细胞(NCC)的迁移。通过胞质C-末端PDZ结合域，ePhin-B配体也能够转导反向信号。在ewitin-B1的情况下，这种通过PDZ结合域的反向信号对于神经脊细胞的正确迁移和正常的颅面发育至关重要。这个应用程序有两个特定的目标。首先，我建议使用免疫沉淀法在NCC中识别和表征颅面发育过程中ePhin-B1反向信号的介体。其次，我建议通过在原代NCC培养中使用可溶性EphB2-Fc融合受体诱导反向信号，并比较表达谱，来鉴定和功能表征ePhin-B1反向信号的转录靶点。这项应用的目的是了解在颅面发育过程中ePhin-B1反向信号的分子机制。

DESCRIPTION (provided by applicant): The mechanisms controlling neural crest migration during craniofacial development are not well understood. The Eph/Ephrin cell signaling receptor-ligand pair has been identified as playing crucial roles during development, including controlling neural crest cell (NCC) migration. The ephrin-B ligands are also capable of transducing reverse signaling by virtue of their cytoplasmic C-terminal PDZ binding domain. In the case of ephrin-B1, this reverse signaling via the PDZ-binding domain is crucial for the proper migration of neural crest cells and normal craniofacial development. There are two specific aims for this application. First, I propose to identify and characterize mediators of ephrin-B1 reverse signaling during craniofacial development using an immunoprecipitation approach in NCCs. Second, I propose to identify and functionally characterize transcriptional targets of ephrin-B1 reverse signaling by inducing reverse signaling using a soluble EphB2-Fc fusion receptor in primary NCC culture and comparing expression profiles. The goal of this application is to understand molecular mechanisms of ephrin-B1 reverse signaling during craniofacial development.