Production of laminin and fibronectin by Schwannoma cells: cell-protein interactions in vitro and protein localization in peripheral nerve in vivo.
Production of laminin and fibronectin by Schwannoma cells: cell-protein interactions in vitro and protein localization in peripheral nerve in vivo.
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DOI：
10.1083/jcb.96.5.1218
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发表时间：
1983-05
期刊：
影响因子：
--
通讯作者：
Furcht LT
中科院分区：
文献类型：
--
作者：
Palm SL;Furcht LT
我们研究了大鼠神经鞘瘤细胞系（RN22F），以确定它是否产生基底膜糖蛋白层粘连蛋白和纤维连接蛋白，以及它如何在体外与这些蛋白相互作用。用抗血清对层粘连蛋白和纤连蛋白进行免疫沉淀实验和电镜水平的免疫细胞化学定位。条件培养基和溶解的神经鞘瘤细胞的抗层粘连蛋白免疫沉淀物的聚丙烯酰胺凝胶含有降低的Mr为20万和15万的条带。条件培养基的抗层粘连蛋白免疫沉淀含有85万道尔顿和15万道尔顿的非还原带，溶解细胞的免疫沉淀含有85万道尔顿、40万道尔顿、20万道尔顿和15万道尔顿的非还原带。条件培养基的抗纤连蛋白免疫沉淀含有22万道尔顿的还原带，44万道尔顿和22万道尔顿的非还原带。在溶解细胞的抗纤连蛋白免疫沉淀物中未检测到放射性标记蛋白。免疫细胞化学发现层粘连蛋白沿细胞表面呈连续带状分布，而纤维连接蛋白仅沿细胞表面稀疏分布。在细胞粘附试验中，与纤维连接蛋白或未包被的底物相比，神经鞘瘤细胞优先结合层粘连蛋白包被的底物。许多神经鞘瘤细胞在层粘连蛋白底物上表现为弯曲和细长的形态，而在纤维连接蛋白底物上则表现为均匀分布的形态，而在非涂层底物上则表现为圆形、非扩散的形态。免疫荧光染色显示，小鼠坐骨神经的层粘连蛋白主要存在于神经内膜和神经会膜中，纤维连接蛋白主要存在于神经会膜中。雪旺细胞产生层粘连蛋白和纤维连接蛋白可能在周围神经的发育和髓鞘形成以及神经损伤后轴突的适当再生中起重要作用。
We studied a rat Schwannoma cell line (RN22F) to determine if it produced the basement membrane glycoproteins laminin and fibronectin, and how it interacted with these proteins in vitro. We used antisera to laminin and fibronectin for immunoprecipitation experiments and immunocytochemical localization at the electron microscope level. Polyacrylamide gels of antilaminin immunoprecipitates of conditioned medium and solubilized Schwannoma cells contained bands of reduced Mr 200,000 and 150,000. Antilaminin immunoprecipitates of conditioned medium contained nonreduced bands of 850,000 daltons and 150,000, and immunoprecipitates of solubilized cells contained nonreduced bands of 850,000, 400,000, 200,000, and 150,000 daltons. Antifibronectin immunoprecipitates of conditioned medium contained a reduced band of 220,000 daltons, and nonreduced bands of 440,000 and 220,000 daltons. Radio-labeled protein was not detected in antifibronectin immunoprecipitates of solubilized cells. By immunocytochemistry, laminin was found along the cell surface in a continuous band, whereas fibronectin was only sparsely distributed along the cell surface. In cell adhesion assays, Schwannoma cells bound preferentially to laminin- coated substrates as compared to fibronectin or noncoated substrates. A number of Schwannoma cells displayed a curved and elongated morphology on laminin substrates, as compared to a uniformly spread morphology on fibronectin, and a round, nonspread morphology on noncoated substrates. Immunofluorescent staining showed laminin in the endoneurium and perineurium and fibronectin predominantly in the perineurium of mouse sciatic nerve in vivo. The production of laminin and fibronectin by Schwann cells may be important in the development and myelination of peripheral nerves, and the proper regeneration of axons following nerve injury.