Primary cilia mediate mechanotransduction through control of ATP-induced Ca2+ signaling in compressed chondrocytes
Primary cilia mediate mechanotransduction through control of ATP-induced Ca2+ signaling in compressed chondrocytes
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DOI：
10.1096/fj.11-193649
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发表时间：
2012-04-01
期刊：
影响因子：
4.8
通讯作者：
Knight, Martin M.
中科院分区：
文献类型：
--
作者：
Wann, Angus K. T.;Zuo, Ning;Knight, Martin M.
我们研究了软骨细胞初级纤毛在与软骨细胞外基质合成相关的机械转导事件中的作用。我们产生了条件永生化的野生型（WT）和IFFT 88（orpk）（ORPK）突变软骨细胞，缺乏初级纤毛和评估细胞内Ca 2+信号，细胞外基质合成，ATP释放响应生理相关的压缩应变在三维软骨细胞培养系统。将所有条件与未加载的对照进行比较。我们发现，纤毛所需的压缩诱导的Ca 2+信号转导介导的ATP释放，以及相关的上调聚集蛋白聚糖mRNA和硫酸化糖胺聚糖分泌。然而，软骨细胞纤毛不是最初的机械感受器，因为WT和ORPK细胞都显示出机械诱导的ATP释放。相反，我们发现，初级纤毛下游ATP接收所需的，因为ORPK细胞不引起Ca 2+反应外源性ATP，即使WT和ORPK细胞表达类似水平的嘌呤受体。我们认为，嘌呤能钙信号可能是由多囊蛋白-1，因为ORPK细胞只表达C-末端尾巴。这是第一项证明初级纤毛是软骨机械转导的重要细胞器的研究，并确定了初级纤毛以前在任何其他细胞类型中未报道的新作用，即纤毛介导的ATP接收控制。Wann，A. K. T.，左，N。，海克拉夫特角J.，詹森角G.，普尔角一、McGlashan，S. R.，奈特，M。M.初级纤毛通过控制压缩软骨细胞中ATP诱导的Ca 2+信号传导介导机械转导。FASEB J.26，1663-1671（2012）。www.fasebj.org
We investigated the role of the chondrocyte primary cilium in mechanotransduction events related to cartilage extracellular matrix synthesis. We generated conditionally immortalized wild-type (WT) and IFT88(orpk) (ORPK) mutant chondrocytes that lack primary cilia and assessed intracellular Ca2+ signaling, extracellular matrix synthesis, and ATP release in response to physiologically relevant compressive strains in a 3-dimensional chondrocyte culture system. All conditions were compared to unloaded controls. We found that cilia were required for compression-induced Ca2+ signaling mediated by ATP release, and an associated up-regulation of aggrecan mRNA and sulfated glycosaminosglycan secretion. However, chondrocyte cilia were not the initial mechanoreceptors, since both WT and ORPK cells showed mechanically induced ATP release. Rather, we found that primary cilia were required for downstream ATP reception, since ORPK cells did not elicit a Ca2+ response to exogenous ATP even though WT and ORPK cells express similar levels of purine receptors. We suggest that purinergic Ca2+ signaling may be regulated by polycystin-1, since ORPK cells only expressed the C-terminal tail. This is the first study to demonstrate that primary cilia are essential organelles for cartilage mechanotransduction, as well as identifying a novel role for primary cilia not previously reported in any other cell type, namely cilia-mediated control of ATP reception.-Wann, A. K. T., Zuo, N., Haycraft, C. J., Jensen, C. G., Poole, C. A., McGlashan, S. R., Knight, M. M. Primary cilia mediate mechanotransduction through control of ATP-induced Ca2+ signaling in compressed chondrocytes. FASEB J. 26, 1663-1671 (2012). www.fasebj.org