Investigation of human keratinocyte cell adhesion using atomic force microscopy
Investigation of human keratinocyte cell adhesion using atomic force microscopy
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DOI：
10.1016/j.nano.2009.05.008
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发表时间：
2010-02-01
影响因子：
5.4
通讯作者：
Sinha, Animesh A.
中科院分区：
文献类型：
--
作者：
Fung, Carmen Kar Man;Seiffert-Sinha, Kristina;Sinha, Animesh A.
桥粒连接是上皮组织内细胞黏附所必需的特殊结构。在诸如天疱疮等水疱性皮肤病中，由于产生了针对特定桥粒蛋白的自身抗体，可观察到这些连接的解体。然而，在正常生理和疾病条件下有关细胞连接活动的许多细节仍有待阐明。由于其复杂的结构，桥粒连接不太适合用于高分辨率三维结构 - 功能分析的现有技术。在此，原子力显微镜（AFM）被用于对人上皮细胞的细胞连接进行详细的表征和可视化。我们证明了能够在高放大倍数下对细胞连接的详细三维结构进行成像。此外，在抗体处理前后的纵向分析中研究了特异性抗体与细胞连接的桥粒成分结合的效果。我们表明，针对桥粒芯糖蛋白3（桥粒结构单元的主要成分，也是寻常型天疱疮患者自身抗体的主要靶点）的抗体与人角质形成细胞的细胞表面变化以及角质形成细胞间黏附结构的改变有关，这支持了抗体结合会改变细胞结构和连接的论断。本研究表明，通过原子力显微镜可以更全面地分析和表征间隙连接的分子结构，为更好地理解水疱性皮肤病的疾病机制并潜在地监测治疗策略提供了一种新的技术方法。
临床编辑按：在天疱疮等水疱性皮肤病中可观察到桥粒连接的解体。本研究表明，通过原子力显微镜可以更全面地分析和表征间隙连接的分子结构。由爱思唯尔公司出版。
Desmosomal junctions are specialized structures critical to cellular adhesion within epithelial tissues. Disassembly of these junctions is seen consequent to the development of autoantibodies directed at specific desmosomal proteins in blistering skin diseases such as pemphigus. However, many details regarding cell junction activity under normal physiological and disease conditions remain to be elucidated. Because of their complex structure, desmosomal junctions are not well suited to existing techniques for high-resolution three-dimensional structure-function analyses. Here, atomic force microscopy (AFM) is used for detailed characterization and visualization of the cell junctions of human epithelial cells. We demonstrate the ability to image the detailed three-dimensional structure of the cell junction at high magnification. In addition, the effect of specific antibody binding to desmosomal components of the cell junction is studied in longitudinal analyses before and after antibody treatment. We show that antibodies directed against desmoglein 3 (a major component of the desmosomal structural unit, and the major target of autoantibodies in patients with pemphigus vulgaris) are associated with changes at the cell surface of the human keratinocytes and alterations within keratinocyte intercellular adhesion structures, supporting the assertion that cell structures and junctions are modified by antibody binding. The present study indicates that the molecular structure of gap junctions can be more completely analyzed and characterized by AFM, offering a new technological approach to facilitate a better understanding of disease mechanisms and potentially monitor therapeutic strategies in blistering skin diseases.From the Clinical Editor: Disassembly of desmosomal junctions is seen in blistering skin diseases such as Pemphigus. This present study demonstrates that the molecular structure of gap junctions can be more completely analyzed and characterized by atomic force microscopy. Published by Elsevier Inc.