Dziadek M, Edgar D, Paulsson M, Timpl R, Fleischmajer R
Ann N Y Acad Sci. 1986;486:248-59. doi: 10.1111/j.1749-6632.1986.tb48078.x.
Mouse Schwann cells and rat RN22 schwannoma cells cultured in the absence of neurons and fibroblasts produce typical basement membrane proteins. Heparan sulfate proteoglycan (low density form), nidogen, and protein BM-40 were identified by radioimmunoassays, immunoblotting, and by immunoprecipitation after metabolic labeling. The cells also produce a laminin-like protein that differs from authentic laminin by a reduced A chain content and lack of antigenic determinants located in the long arm of laminin. Laminin possessing A and B chains is, however, produced by PYS-2 teratocarcinoma cells grown under the same conditions. Laminin from Schwann cell culture medium promotes neurite outgrowth, and this activity could be immunoprecipitated but not blocked by various antibodies against authentic laminin. In addition, Schwann cell laminin is found complexed noncovalently with nidogen. Sulfate incorporation revealed the synthesis of proteoglycans and entactin. A similar set of proteins and in addition collagen IV could be demonstrated in neurofibroma tissue by immunohistology, and were localized to the laminae densae of the multilayered basement membranes around Schwann cells and capillaries. Laminin purified from 0.5 M NaCl neurofibroma tissue extracts possessed both A and B chains. Nidogen was identified in a partially degraded form.
在没有神经元和成纤维细胞的情况下培养的小鼠雪旺细胞和大鼠RN22雪旺瘤细胞会产生典型的基底膜蛋白。通过放射免疫测定、免疫印迹以及代谢标记后的免疫沉淀法鉴定出硫酸乙酰肝素蛋白聚糖(低密度形式)、巢蛋白和蛋白BM - 40。这些细胞还产生一种类层粘连蛋白,其A链含量减少且缺乏位于层粘连蛋白长臂上的抗原决定簇,与真正的层粘连蛋白不同。然而,在相同条件下生长的PYS - 2畸胎癌细胞会产生具有A链和B链的层粘连蛋白。雪旺细胞培养基中的层粘连蛋白可促进神经突生长,这种活性可以通过免疫沉淀法检测到,但不能被针对真正层粘连蛋白的各种抗体阻断。此外,发现雪旺细胞层粘连蛋白与巢蛋白非共价结合。硫酸盐掺入显示蛋白聚糖和内动蛋白的合成。通过免疫组织学可以在神经纤维瘤组织中证明一组类似的蛋白质,此外还有IV型胶原蛋白,它们定位于雪旺细胞和毛细血管周围多层基底膜的致密层中。从0.5M NaCl神经纤维瘤组织提取物中纯化的层粘连蛋白同时具有A链和B链。巢蛋白以部分降解的形式被鉴定出来。