Department of Pharmaceutical Sciences, Northeastern Universities Colleges of Medicine and Pharmacy, Rootstown, Ohio 44272, USA.
J Cell Physiol. 2012 Jan;227(1):70-6. doi: 10.1002/jcp.22704.
Cilia are mechanosensing organelles that communicate extracellular signals into intracellular responses. Altered functions of primary cilia play a key role in the development of various diseases including polycystic kidney disease. Here, we show that endothelial cells from the oak ridge polycystic kidney (Tg737(orpk/orpk) ) mouse, with impaired cilia assembly, exhibit a reduction in the actin stress fibers and focal adhesions compared to wild-type (WT). In contrast, endothelial cells from polycystin-1 deficient mice (pkd1(null/null) ), with impaired cilia function, display robust stress fibers, and focal adhesion assembly. We found that the Tg737(orpk/orpk) cells exhibit impaired directional migration and endothelial cell monolayer permeability compared to the WT and pkd1(null/null) cells. Finally, we found that the expression of heat shock protein 27 (hsp27) and the phosphorylation of focal adhesion kinase (FAK) are downregulated in the Tg737(orpk/orpk) cells and overexpression of hsp27 restored both FAK phosphorylation and cell migration. Taken together, these results demonstrate that disruption of the primary cilia structure or function compromises the endothelium through the suppression of hsp27 dependent actin organization and focal adhesion formation, which may contribute to the vascular dysfunction in ciliopathies.
纤毛是一种机械感受器,可将细胞外信号转导为细胞内反应。初级纤毛功能异常在包括多囊肾病在内的多种疾病的发生发展中起关键作用。在这里,我们发现与野生型(WT)相比,组装受损的 Oak Ridge 多囊肾病(Tg737(orpk / orpk))小鼠的内皮细胞中肌动蛋白应力纤维和焦点黏附减少。相比之下,纤毛功能受损的多囊蛋白-1 缺陷型(pkd1(null / null))小鼠的内皮细胞显示出强大的应力纤维和焦点黏附组装。我们发现与 WT 和 pkd1(null / null)细胞相比,Tg737(orpk / orpk)细胞表现出受损的定向迁移和内皮细胞单层通透性。最后,我们发现 Tg737(orpk / orpk)细胞中热休克蛋白 27(hsp27)的表达和焦点黏附激酶(FAK)的磷酸化下调,而过表达 hsp27 恢复了 FAK 磷酸化和细胞迁移。总之,这些结果表明,初级纤毛结构或功能的破坏通过抑制 hsp27 依赖性肌动蛋白组织和焦点黏附形成来损害内皮细胞,这可能导致纤毛病中的血管功能障碍。