Mechanisms of Cell-Matrix Interaction and Signaling in Lens Development

失去细胞基质黏附部位和细胞骨架之间的关键连接可能是 一些疾病。这个项目的重点是阐明调控这一过程的分子机制。 在发育中的小鼠晶状体中的附着物。我们的中心假设是粘着斑激酶 FAK)和相关的信号分子将细胞-基质附着转化为细胞骨架重排和 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。对这种动态的、双向的研究 交互作用需要检查天然的三维组织结构。我们的总体战略是 在小鼠晶状体中研究这些通路，因为它被最厚的基底层之一所包围 体内的膜经过一定的发育，可以原封不动地培养，并且是透明的， 使其成为成像分析的唯一可访问对象。我们将使用有条件的淘汰法删除 FAK，其唯一的家族成员Pyk2，以及来自发育中的小鼠晶状体的整合素连接激酶(ILK)。 删除Pyk2将解决功能冗余的问题，而删除ilk将进一步考验我们的 通过另一种途径破坏细胞-基质-细胞骨架连接的假说，独立于 FAK。我们将使用分子、细胞、生化和结构方法的组合来解决 以下问题：目的1)晶状体发育是否需要FAK、PYK2和ILK信号？会吗？ 这一通路的紊乱会导致晶状体病理吗？目标2)中断FAK相关的信令结果 晶状体细胞迁移、增殖和/或分化中的特定细胞缺陷？哪些信号通路 有牵连吗？目的3)FAK相关信号的中断是如何改变晶状体囊膜基底的 膜是有组织的、合成的和重新建模的吗？这些拟议的研究将提供机械性的 深入了解细胞和细胞外基质之间的串扰，以及信号通路 牵涉其中。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 并有可能更好地了解致盲疾病，如小眼炎和 我们预测的先天性白内障可能是由于晶状体细胞过程中细胞基质相互作用错误所致 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.