Glomerular Cell Cross-Talk Influences Composition and Assembly of Extracellular Matrix
Glomerular Cell Cross-Talk Influences Composition and Assembly of Extracellular Matrix
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DOI：
10.1681/asn.2013070795
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发表时间：
2014-05-01
影响因子：
13.6
通讯作者：
Lennon, Rachel
中科院分区：
文献类型：
--
作者：
Byron, Adam;Randles, Michael J.;Lennon, Rachel
肾小球基底膜 (GBM) 是肾小球内特殊的细胞外基质 (ECM) 隔室，其中含有组织限制的 IV 型胶原蛋白和层粘连蛋白同工型。它是毛细血管壁的组成部分，因此在功能上与肾小球滤过相关。尽管已经通过全局和基于候选的方法研究了 GBM 的组成，但肾小球细胞类型对 ECM 产生的相对贡献尚不清楚。为了表征细胞对 GBM 的特定贡献，我们使用基于质谱的蛋白质组学来分析体外从足细胞和肾小球内皮细胞分离的 ECM。这些分析确定了 ECM 组成中细胞类型特异性的差异，表明对肾小球 ECM 组装的不同贡献。与单一培养的 ECM 相比，足细胞和内皮细胞的共培养导致 ECM 的组成和组织发生改变，电子显微镜显示共培养的细胞之间存在基底膜样 ECM 沉积，表明细胞间的串扰参与了肾小球 ECM 的产生。值得注意的是，与单一培养 ECM 蛋白质组相比，共培养 ECM 蛋白质组更类似于组织来源的肾小球 ECM 数据集，表明其与体内 GBM 的相关性。蛋白质网络分析揭示了 35 种高度连接的结构 ECM 蛋白的共同核心，这可能对肾小球 ECM 组装很重要。总体而言，这些发现显示了肾小球 ECM 的复杂性，并表明 ECM 的组成和组织都依赖于环境。
The glomerular basement membrane (GBM) is a specialized extracellular matrix (ECM) compartment within the glomerulus that contains tissue-restricted isoforms of collagen IV and laminin. It is integral to the capillary wall and therefore, functionally linked to glomerular filtration. Although the composition of the GBM has been investigated with global and candidate-based approaches, the relative contributions of glomerular cell types to the production of ECM are not well understood. To characterize specific cellular contributions to the GBM, we used mass spectrometry-based proteomics to analyze ECM isolated from podocytes and glomerular endothelial cells in vitro. These analyses identified cell type-specific differences in ECM composition, indicating distinct contributions to glomerular ECM assembly. Coculture of podocytes and endothelial cells resulted in an altered composition and organization of ECM compared with monoculture ECMs, and electron microscopy revealed basement membrane-like ECM deposition between cocultured cells, suggesting the involvement of cell-cell cross-talk in the production of glomerular ECM. Notably, compared with monoculture ECM proteomes, the coculture ECM proteome better resembled a tissue-derived glomerular ECM dataset, indicating its relevance to GBM in vivo. Protein network analyses revealed a common core of 35 highly connected structural ECM proteins that may be important for glomerular ECM assembly. Overall, these findings show the complexity of the glomerular ECM and suggest that both ECM composition and organization are context-dependent.