ADAM FUNCTION IN NEURAL CREST CELLS

描述（由申请人提供）：颅神经嵴（CNC）是一个 细胞群形成于前神经和 脊椎动物胚胎中的非神经外胚层。这些细胞经历了一个受调节的 上皮-间充质转化，从发育中的神经管出现， 沿着沿着限定的路径在整个头部迁移。CNC细胞产生一种 多种细胞类型和组织，包括软骨和骨骼成分 头和脸。诱导、增殖、迁移和/或 CNC细胞的分化解释了广泛的颅面 异常因此，对指导CNC的分子事件的基本理解 细胞发育和迁移行为对于理解 颅面缺陷的症状ADAM 13是亚当斯家族的成员， 跨膜糖蛋白，包含A去整合素和A金属蛋白酶 域这种蛋白质最初是在非洲爪蟾中发现的， 在CNC细胞迁移之前和迁移过程中表达。ADAM 13 金属蛋白酶活性被假设为在 通过修饰细胞外基质（ECM）来改变CNC的迁移行为 这些细胞通过的填充空间。因此整体 本研究的目的是确定ADAM 13和其他 在神经嵴中表达的ADAM家族成员。主 目的是建立ADAM 13参与CNC迁移的机制。 将使用胚胎转录本注射、转基因方法和转染 表达ADAM 13野生型和突变型构建体，以研究ADAM 13在完整的非洲爪蟾胚胎、培养的外植体和单细胞中的功能。的 单个ADAM 13结构域的功能特异性（即，金属蛋白酶， 去整合素、富含半胱氨酸和细胞质）也将使用嵌合的 由ADAM 13和其他结构相关的片段组成的ADAM构建体 具有不同生物活性的亚当斯。ADAM 13胞质尾区具有 已显示与含SH 3结构域的蛋白质相互作用，包括Src 1和 PACSIN-2。PACSIN-2的结合也与ADAM 13的减少有关。 体内功能活性。因此，拟议研究的最终目的是 研究PACSIN-2下调ADAM 13功能的机制。

DESCRIPTION (provided by applicant): The cranial neural crest (CNC) is a population of cells formed at the border between the anterior neural and non-neural ectoderm in vertebrate embryos. These cells undergo a regulated epithelial-mesenchymal transition, emerging from the developing neural tube and migrating throughout the head along defined pathways. CNC cells give rise to a variety of cell types and tissues including the cartilage and skeletal elements of the head and face. Defects in the induction, proliferation, migration and/or differentiation of CNC cells account for a wide range of craniofacial anomalies. Thus, a basic understanding of the molecular events that direct CNC cell development and migratory behavior is critical to understanding the causes of craniofacial defects. ADAM 13 is a member of the ADAMs family of transmembrane glycoproteins, which contain A Disintegrin and A Metalloprotease domain. This protein was identified originally in Xenopus laevis, where it is expressed in CNC cells before and during their migration. ADAM 13 metalloprotease activity is hypothesized to play a critical role in the migratory behaviors of the CNC by modifying the extracellular matrix (ECM) filled spaces through which these cells travel. Therefore, the overall objectives of this study are to establish the functions of ADAM 13 and other members of the ADAM family that are expressed in the neural crest. A primary goal is to establish the mechanism of ADAM 13 involvement in CNC migration. Embryo transcript injection, transgenic methods and transfection will be used to express ADAM 13 wild type and mutant constructs in order to investigate ADAM 13 function in intact Xenopus embryos, cultured explants and single cells. The functional specificities of individual ADAM 13 domains (i.e., metalloprotease, disintegrin, cysteine-rich and cytoplasmic) will also be studied using chimeric ADAM constructs comprised of segments of ADAM 13 and other structurally related ADAMs with differing biological activities. The ADAM 13 cytoplasmic tail has been shown to interact with SH3 domain containing proteins including Src1 and PACSIN-2. PACSIN-2 binding is also associated with a reduction in ADAM 13 functional activity in vivo. Thus, the final aim of the proposed research is to investigate the mechanism by which PACSIN-2 down-regulates ADAM 13 function.