IDENTIFICATION OF INTEGRIN CELL SUBSTRATUM ADHESION RECEPTORS ON CULTURED RAT BONE-CELLS
IDENTIFICATION OF INTEGRIN CELL SUBSTRATUM ADHESION RECEPTORS ON CULTURED RAT BONE-CELLS
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DOI：
10.1002/jor.1100100604
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发表时间：
1992-11-01
影响因子：
2.8
通讯作者：
ALBELDA, SM
中科院分区：
文献类型：
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作者：
BRIGHTON, CT;ALBELDA, SM
骨细胞与其细胞外基质的相互作用在骨发育、修复和疾病中具有重要意义。我们研究了大鼠颅骨骨细胞粘附各种基质蛋白的能力，并确定了整合素细胞基质粘附受体在这些相互作用中的作用。用H-3-胸苷预标记分离的新生大鼠颅骨骨细胞，并将其铺在已预涂有各种底物系列稀释液的塑料威尔斯孔上，显示出对纤连蛋白、纤维蛋白原、层粘连蛋白、玻连蛋白和胶原I和IV的典型剂量-反应粘附曲线。在>0.0001 μ g/ml的所有底物浓度下，细胞对多聚-D-赖氨酸（一种非特异性细胞粘附物）的粘附性都很高。多克隆抗大鼠整联蛋白抗体阻断细胞粘附到除聚-D-赖氨酸之外的所有测试底物。分离的大鼠颅骨骨细胞用I-125表面标记，提取，并用针对大鼠整联蛋白复合物和衍生自α 2、α 3和α 5亚基的胞质结构域的肽制备的多克隆抗体免疫沉淀。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（非还原）分析，鉴定出4条代表整合素混合物的条带，包括α 1 β 1层粘连蛋白/胶原蛋白受体、α 5 β 1纤连蛋白受体和α（v）β 3（或可能的α（v）β 5）玻连蛋白受体。这些实验表明，骨细胞通过特异性整合素粘附于多种细胞外基质蛋白。增加对这些受体的调节以及它们向细胞传递信息的机制的了解对于更全面地理解骨生理学和病理生理学将是重要的。
The interactions of bone cells with their extracellular matrix is of major importance in bone development, repair, and discase. We examined the ability of rat calvarial bone cells to adhere to various matrix proteins and to define the role of integrin cell-substrate adhesion receptors in these interactions. Isolated newborn rat calvarial bone cells prelabeled with H-3-thymidine and plated on plastic wells that had been precoated with serial dilutions of various substrates showed typical dose-response adherence curves to fibronectin, fibrinogen, laminin, vitronectin, and collagen I and IV. Cell adherence to poly-D-lysine, a nonspecific cell adherent, was high at all substrate concentrations >0.0001 mug/ml. A polyclonal anti-rat integrin antibody blocked cell adhesion to all substrates tested except poly-D-lysine. Isolated rat calvarial bone cells were surface labeled with I-125, extracted, and immunoprecipitated with polyclonal antibodies made against the rat integrin complex and peptides derived from the cytoplasmic domains of the alpha2, alpha3, and alpha5 subunits. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (nonreduced) identified four bands representing a mixture of integrins including the alpha1beta1 laminin/collagen receptor, the alpha5beta1 fibronectin receptor, and the alpha(v)beta3 (or possibly alpha(v)beta5) vitronectin receptor. These experiments show that bone cells adhere to a wide variety of extracellular matrix proteins via specific integrins. Increased knowledge about the regulation of these receptors and the mechanisms by which they transmit information to the cell will be important for a more complete understanding of bone physiology and pathophysiology.