Mechanisms of Cell-Matrix Interaction and Signaling in Lens Development

细胞基质粘附位点和细胞骨架之间的关键连接的丧失可能是细胞凋亡的核心。 一些疾病。该项目的重点是阐明调控这一过程的分子机制。 显影鼠标透镜中的摩擦附件。我们的中心假设是黏着斑激酶 （FAK）和相关信号分子将细胞-基质附着转化为细胞骨架重排， 一系列反应，包括1）细胞增殖、分化和迁移，以及2） 将细胞外基质组织成基底膜。研究这种动态的、双向的 相互作用需要检查天然的三维组织结构。我们的总体战略是 研究小鼠眼透镜中的这些通路，因为它被最厚的基底之一所包围。 膜在体内，经历了一个明确的发展，可以培养完整，是透明的， 使其唯一地可用于成像分析。我们将使用条件淘汰法删除 FAK、其唯一的家族成员Pyk 2以及来自发育中的小鼠透镜的整合素连接激酶（ILK）。 删除Pyk 2将解决功能冗余的问题，删除ILK将进一步测试我们的 假设通过另一种途径破坏细胞-基质-细胞骨架附着， 假的。我们将使用分子、细胞、生物化学和结构方法的组合来解决这些问题。 以下问题：目的1）FAK、Pyk 2和ILK信号传导是透镜发育所必需的吗？并 该通路扰动会导致透镜病变吗？目的2）FAK相关信号传导的中断是否导致 在透镜细胞迁移、增殖和/或分化的特定细胞缺陷中？什么信号通路 有关系吗目的3）FAK相关信号的破坏如何改变透镜囊基底 膜是组织的，合成的，重新建模的？这些拟议的研究将提供机制 深入了解细胞和细胞外基质之间所需的串扰，以及信号通路 涉案重要的是，这些结果还将为眼睛发育的机制提供独特的见解 并可能有助于更好地了解致盲性疾病，如小眼症， 我们预测先天性白内障可能是由于透镜细胞生长过程中细胞基质相互作用的缺陷引起的。 发展

Loss of the pivotal connection between cell matrix adhesion sites and the cytoskeleton may lie at the heart of a number of diseases. This project is focused on elucidating the molecular mechanisms regulating this Imctional attachment in the developing mouse lens. Our central hypothesis is that focal adhesion kinase ^FAK) and related signaling molecules translate cell-matrix attachment into cytoskeletal rearrangements and a cascade of responses, including 1) cellular proliferation, differentiation and migration as well as 2) organization of the extracellular matrix into basement membranes. Study of this dynamic, bi-directional nteraction requires examination of a native, three-dimensional tissue structure. Our general strategy is to study these pathways in the mouse ocular lens, since it is surrounded by one of the thickest basement membranes in the body, undergoes a defined development, can be cultured intact, and is transparent, rendering it uniquely accessible to imaging analysis. We will use a conditional knockout approach to delete FAK, its only family member Pyk2, as well as integrin-linked kinase (ILK) from the developing mouse lens. Deletion of Pyk2 will addresses issues of functional redundancy, and deletion of ILK will further test our hypothesis by disrupting cell-matrix-cytoskeletal attachment through an alternative route, independent of FAK. We will use a combination of molecular, cellular, biochemical and structural approaches to address the following questions: Aim 1) Are FAK, Pyk2 and ILK signaling required for lens development? Does perturbation of this pathway result in lens pathology? Aim 2) Does disruption of FAK-related signaling result in specific cellular defects in lens cell migration, proliferation and/or differentiation? What signaling pathways are involved? Aim 3) How does disruption of FAK-related signaling alter how the lens capsule basement membrane is organized, synthesized and re-modeled? These proposed studies will provide mechanistic insight into the crosstalk required between cells and the extracellular matrix, and the signaling pathways involved. Importantly, these results will also provide unique insight into the mechanisms of eye development and potentially contriubte to a better understanding of blinding disorders such as microphthalmia and congenital cataracts that we predict may result from faulty cell matrix interactions during lens cell development.