Broussard Joshua A, Diggins Nicole L, Hummel Stephen, Georgescu Walter, Quaranta Vito, Webb Donna J
Department of Biological Sciences and Vanderbilt Kennedy Center for Research on Human Development, Vanderbilt University, Nashville, Tennessee 37235.
Center for Cancer Systems Biology at Vanderbilt, Vanderbilt University, Nashville, Tennessee 37235.
Sci Rep. 2015 Jan 29;5:8124. doi: 10.1038/srep08124.
Cell-matrix adhesions are of great interest because of their contribution to numerous biological processes, including cell migration, differentiation, proliferation, survival, tissue morphogenesis, wound healing, and tumorigenesis. Adhesions are dynamic structures that are classically defined on two-dimensional (2D) substrates, though the need to analyze adhesions in more physiologic three-dimensional (3D) environments is being increasingly recognized. However, progress has been greatly hampered by the lack of available tools to analyze adhesions in 3D environments. To address this need, we have developed a platform for the automated analysis, segmentation, and tracking of adhesions (PAASTA) based on an open source MATLAB framework, CellAnimation. PAASTA enables the rapid analysis of adhesion dynamics and many other adhesion characteristics, such as lifetime, size, and location, in 3D environments and on traditional 2D substrates. We manually validate PAASTA and utilize it to quantify rate constants for adhesion assembly and disassembly as well as adhesion lifetime and size in 3D matrices. PAASTA will be a valuable tool for characterizing adhesions and for deciphering the molecular mechanisms that regulate adhesion dynamics in 3D environments.
细胞与基质的黏附备受关注，因为它们对众多生物学过程有重要作用，包括细胞迁移、分化、增殖、存活、组织形态发生、伤口愈合和肿瘤发生。黏附是动态结构，传统上是在二维（2D）基质上定义的，不过人们越来越认识到需要在更接近生理状态的三维（3D）环境中分析黏附。然而，由于缺乏在3D环境中分析黏附的可用工具，进展受到了极大阻碍。为满足这一需求，我们基于开源MATLAB框架CellAnimation开发了一个用于黏附自动分析、分割和跟踪的平台（PAASTA）。PAASTA能够在3D环境和传统2D基质上快速分析黏附动力学以及许多其他黏附特性，如寿命、大小和位置。我们手动验证了PAASTA，并利用它来量化3D基质中黏附组装和解聚的速率常数以及黏附寿命和大小。PAASTA将成为表征黏附以及解读在3D环境中调节黏附动力学的分子机制的宝贵工具。
