Mapping the mechanics and macromolecular organization of hyaluronan-rich cell coats
Mapping the mechanics and macromolecular organization of hyaluronan-rich cell coats
复制标题
DOI：
10.1039/b905574f
复制
发表时间：
2009-01-01
期刊：
影响因子：
3.4
通讯作者：
Curtis, Jennifer E.
中科院分区：
文献类型：
--
作者：
Boehm, Heike;Mundinger, Tabea A.;Curtis, Jennifer E.
富含透明质酸（HA）的细胞外膜（PCC）包裹着大多数哺乳动物细胞，在细胞粘附、增殖、运动和胚胎发生等生物过程中起着至关重要的作用。特别是它在软骨细胞上的存在，它生活在承重软骨中，在骨关节炎等疾病中具有广泛的意义，突出了它在生物体中的机械作用。尽管具有重要意义，但细胞外壳的大分子组织仍然是推测性的。为了获得高度水合PCCs的更详细的空间图像，我们提出了两种独立但互补的非侵入性技术，用于细胞外壳的机械和结构特性的位置分辨分析。位置依赖的微流变学提供了PCC的微力学图，揭示了模型大鼠软骨细胞（RCJ-P）向质膜增加弹性刚度的梯度。这种梯度可以与透明质酸的相对分布相关，这是通过egfp标记的神经能结合结构域推断出来的，这是一种与透明质酸结合的小荧光分子。HA浓度分布的空间变化与位置依赖的弹性一致。将这些方法结合起来，可以揭示PCC的分子结构。
The hyaluronan (HA)-rich pericellular coat (PCC) enveloping most mammalian cells plays a vital role in biological processes such as cell adhesion, proliferation, motility and embryogenesis. In particular its presence on chondrocytes, which live in the load-bearing cartilage, has a wide range of implications in diseases such as osteoarthritis, highlighting its mechanical role in living organisms. Despite its significance, the macromolecular organization of the cell coat remains speculative. In order to obtain a more detailed spatial picture of highly hydrated PCCs, we present two independent but complementary non-invasive techniques for the position-resolved analysis of the cell coat's mechanical and structural properties. Position-dependent microrheology provides a micromechanical map of the PCC that reveals a gradient of increasing elastic stiffness towards the plasma membrane on model rat chondrocyte cells (RCJ-P). This gradient can be correlated with the relative distribution of HA, which is inferred using an eGFP-labelled neurocan-binding domain, a small fluorescent molecule that binds to HA. The spatial variation of the HA concentration profile is consistent with the position-dependent elasticity. Combining these approaches sheds light on the molecular architecture of the PCC.