Podocytes require the engagement of cell surface heparan sulfate proteoglycans for adhesion to extracellular matrices
Podocytes require the engagement of cell surface heparan sulfate proteoglycans for adhesion to extracellular matrices
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DOI：
10.1038/ki.2010.136
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发表时间：
2010-12-01
影响因子：
19.6
通讯作者：
McCarthy, Kevin
中科院分区：
文献类型：
--
作者：
Chen, Shoujun;Wassenhove-McCarthy, Deborah;McCarthy, Kevin
足细胞通过细胞表面受体附着在肾小球基底膜上。由于在其他细胞中，这些粘附被细胞表面蛋白聚糖增强，我们通过培养具有（对照）或不具有（突变）硫酸肝素糖胺聚糖链的永生化足细胞细胞系，研究了这些分子及其糖胺聚糖侧链对足细胞粘附的贡献。在粘附实验中，与突变体相比，控制足细胞的附着、扩散和迁移更有效，表明在这些过程中需要硫酸肝素链。蛋白聚糖syndecan-4已知对细胞附着、扩散和细胞骨架组织有直接影响。我们发现它定位于与应力纤维形成一致的对照足细胞的局灶粘连。在突变细胞中，syndecan-4与较小的局灶接触和皮质肌动蛋白组织有关。流式细胞术分析显示，突变细胞表面syndecan-4的含量是对照细胞的两倍。蛋白激酶C α是一种与syndecan-4结合并被其激活的信号分子，与对照细胞相比，其膜定位激活增加了四倍。在体内，PEXTKO小鼠体内硫酸肝素糖胺聚糖的缺失导致肾小球syndecan-4的缺失。总之，我们的研究进一步证明了细胞表面硫酸肝素糖胺聚糖在足细胞活性中的动态作用。肾脏国际（2010）78,1088-1099；doi: 10.1038 / ki.2010.136;2010年5月12日在线发布
Podocytes adhere to the glomerular basement membrane by cell surface receptors. Since in other cells these adhesions are enhanced by cell surface proteoglycans, we examined the contribution of these molecules and their glycosaminoglycan side chains to podocyte adhesion by developing immortalized podocyte cell lines with (control) or without (mutant) heparan sulfate glycosaminoglycan chains. In adhesion assays control podocytes attached, spread, and migrated more efficiently compared with mutants, indicating a requirement for heparan sulfate chains in these processes. The proteoglycan syndecan-4 is known to have direct effects on cell attachment, spreading, and cytoskeletal organization. We found it localized to focal adhesions in control podocytes coincident with stress fiber formation. In mutant cells, syndecan-4 was associated with smaller focal contacts and cortical actin organization. Analysis by flow cytometry showed that mutant cells had twice the amount of surface syndecan-4 of control cells. Protein kinase C alpha, a signaling molecule bound to and activated by syndecan-4, showed a fourfold increase in membrane localization-activation than that seen in control cells. In vivo, the loss of heparan sulfate glycosaminoglycans in PEXTKO mice led to a loss of glomerular syndecan-4. Overall, our study provides further evidence for a dynamic role of cell surface heparan sulfate glycosaminoglycans in podocyte activity. Kidney International (2010) 78, 1088-1099; doi: 10.1038/ki.2010.136; published online 12 May 2010