ADAM FUNCTION IN NEURAL CREST CELLS

神经嵴细胞中的 ADAM 功能

• 批准号: 6685181
• 负责人: DOUGLAS W. DESIMONE
• 金额: $ 36.88万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-01-01 至 2006-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6685181
• 关键词: Xenopus oocyte; bone development; cell adhesion; cell differentiation; cell migration; cell proliferation; chimeric proteins; craniofacial; cytoskeletal proteins; embryo /fetus cell /tissue; embryogenesis; enzyme activity; enzyme structure; enzyme substrate; extracellular matrix; genetically modified animals; integrins; laboratory mouse; metalloendopeptidases; mutant; neural crest; phosphorylation; protein binding; protein protein interaction; protein structure function; transfection

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.

描述（由申请人提供）：颅神经rest（CNC）是 在前神经和 脊椎动物胚胎中的非神经外胚层。这些细胞经历了受调节的 上皮间质转变，从发育中的神经管中出现 沿着定义的途径在整个头部迁移。 CNC细胞产生 各种细胞类型和组织，包括软骨和骨骼元素 头和脸。诱导，增殖，迁移和/或 CNC细胞的分化构成了广泛的颅面 异常。因此，对引导CNC的分子事件的基本理解 细胞的发展和迁移行为对于理解原因至关重要 颅面缺陷。亚当13是亚当斯家族的成员 跨膜糖蛋白，其中含有分解蛋白和金属蛋白酶 领域。该蛋白最初是在Xenopus laevis中鉴定出来的 在CNC细胞迁移之前和期间在CNC细胞中表达。亚当13 假设金属蛋白酶活性在 CNC的迁移行为通过修改细胞外基质（ECM） 这些单元传播的填充空间。因此，整体 这项研究的目标是建立亚当13和其他其他的功能 在神经座中表达的亚当家族的成员。主要 目标是建立Adam 13参与CNC迁移的机制。 将使用胚胎转录物注射，转基因方法和转染 表达Adam 13野生型和突变体构造以研究亚当 13在完整的爪蟾胚胎，培养的外植体和单个细胞中的功能。这 单个亚当13个域的功能特异性（即金属蛋白酶， 还将使用嵌合体研究解脱蛋白，富含半胱氨酸和细胞质） 亚当的构造由亚当13和其他结构相关的段组成 Adams具有不同的生物学活动。亚当13细胞质尾巴有 已显示与含有蛋白质的SH3结构域相互作用，包括Src1和 Pacsin-2。 PACSIN-2结合也与亚当13的减少有关 体内功能活动。因此，拟议研究的最终目的是 研究PACSIN-2降低ADAM 13功能的机制。